Biomedicine & Pharmacotherapy 112 (2019) 108690

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Biomedicine & Pharmacotherapy

journal homepage: www.elsevier.com/locate/biopha

Cancer cells change their glucose metabolism to overcome increased ROS: T

One step from cancer cell to cancer stem cell?
Zahra Ghanbari Movaheda,1, Mohsen Rastegari-Pouyanib,1, Mohammad hossein Mohammadic,
⁎
Kamran Mansouria,d,
a
Medical Biology Research Center, Kermanshah University of Medical sciences, Kermanshah, Iran
b
Student Research Committee, Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
c
HSCT research center, Laboratory Hematology and blood Banking Department, School of Allied Medical Sciences, Shahid Beheshti University of Medical Science, Tehran,
Iran
d
Department of Molecular Medicine, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran

A R T I C LE I N FO A B S T R A C T

Keywords: Cancer cells can adapt to low energy sources in the face of ATP depletion as well as to their high levels of ROS by
Cancer cell altering their metabolism and energy production networks which might also have a role in determining cell fate
Cancer stem cell and developing drug resistance. Cancer cells are generally characterized by increased glycolysis. This is while;
Glycolysis cancer stem cells (CSCs) exhibit an enhanced pentose phosphate pathway (PPP) metabolism. Based on the
ROS
current literature, we suggest that cancer cells when encountering ROS, first increase the glycolysis rate and then
Pentose phosphate pathway
following the continuation of oxidative stress, the metabolic balance is skewed from glycolysis to PPP. Therefore,
we hypothesize in this review that in cancer cells this metabolic deviation during persistent oxidative stress
might be a sign of cancer cells’ shift towards CSCs, an issue that might be pivotal in more effective targeting of
cancer cells and CSCs.

Abbreviations: ACOX1, acyl-CoA oxidase 1; ACOX2, acyl- CoA oxidase 2; ALD, aldolase; ALDH, aldehyde dehydrogenase; AML, acute myeloid leukemia; AMPK,
AMP-activated protein kinase; ARNT, aryl-hydrocarbon receptor nuclear translocator; ATM, ataxia telangiectasia mutated; BCKDH, Branched-chain alpha-keto acid
dehydrogenase; bHLH-PAS, basic helix-loop-helix/Per-Arnt-Sim; SBPase, sedoheptulose bisphosphatase; CSCs, cancer stem cells; DAO, D-amino acid oxidase; DDO, D-
aspartate oxidase; DHB, ethyl 3,4-dihydroxybenzoic acid; DHODH, dihydroorotate dehydrogenase; DMOG, dimethyloxallyl glycine; DUOX 2, dual oxidase 2; DUOX1,
dual oxidase 1; EMT, epithelial-mesenchymal transition; ENO, enolase; EPO, erythropoietin; ETC, electron transfer chain; ETFQOR, electron-transferring flavoprotein
ubiquinone oxidoreductase; F26-BP, fructose-2, 6-bisphosphate; F6P, fructose-6-phosphate; FA, fatty acid; FDH, folate dehydrogenases; FQR, ferredoxin quinone
reductase; G6P, glucose-6-phosphate; G6PD, glucose-6-phosphate dehydrogenase; GAPDH, Glyceraldehyde-3-phosphate dehydrogenase; GLUTs, glucose transpor-
ters; GPDH, glycerol-3-phosphate dehydrogenase; GPI, glucose-6-phosphate isomerase; GPX, glutathione peroxidase; GSH, glutathione; GSSG, glutathione disulfide;
GST, glutathione S-transferases; H2O2, hydrogen peroxide; HAO1, L-α-hydroxyacidoxidase 1; HAO2, L-α-hydroxyacidoxidase 2; HDAC, histone deacetylase; HIF-1,
hypoxia-inducible factor 1; HK, hexokinase; HMM, human malignant mesothelioma; iBCSCs, induced breast cancer stem cells; IDH, isocitrate dehydrogenases; iNOS,
inducible nitric oxide synthase; LDH, lactate dehydrogenase; LET, low linear energy transfer; MAOs, monoamine oxidases; ME, malic enzymes; mGPDH, mi-
tochondrial glycerol-3-phosphate dehydrogenase; MnSOD, manganese superoxide dismutase; mtDNA, mitochondrial DNA; MTHFD1, methylene tetrahydrofolate
dehydrogenase 1; MTHFD2L, MTHFD2-like; NGF, nerve growth factor; NOS, nitric oxide synthase; NOX1, NADPH oxidase 1; NOX2, NADPH oxidase 2; NOX4,
NADPH oxidase 4; O2%−, superoxide; OGDH, 2-oxoglutarate dehydrogenase; OGD, oxygen-glucose deprivation; OXPHOS, oxidative phosphorylation; PAOX, poly-
amine oxidase; PARP, poly(ADP-ribose) polymerase; PDH, pyruvate dehydrogenase; PDK-1, pyruvate dehydrogenase kinase 1; PEP, phosphoenolpyruvate; PFK1,
phosphofructokinase 1; PFKFB, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases; PG, phosphoglycerate; PGDH, phosphogluconate dehydrogenase; PGK,
phosphoglycerate kinase; PGL, phosphogluconolactonase; PHGDH, phosphoglycerate dehydrogenase; PHP, phosphopyruvate; PIKKs, phosphatidylinositol-3-kinase-
related protein kinases; PIPOX, L-pipecolic acid oxidase; PK, pyruvate kinase; PKL, liver-type pyruvate kinase; PKM2, pyruvate kinase M2; PKR, red blood cell
pyruvate kinase; PPP, pentose phosphate pathway; PSAT1, phosphoserine aminotransferase; P-Ser, phosphoserine; PSPH, phosphoserine phosphatase; RNS, reactive
nitrogen species; ROS, reactive oxygen species; RPE, ribulose 5-phosphate epimerase; RPI, ribose 5-phosphate isomerase; SAHA, suberoylanilide hydroxamic acid;
SHMT2, serine hydroxymethyltransferase
⁎
Corresponding author at: Kamran Mnasouri,Medical Biology Research Center, School of Medicine, Sorkheh Ligeh Blvd, P.O. Box 1568, Kermanshah, Iran.
E-mail address: kmansouri@kums.ac.ir (K. Mansouri).
1
The first two authors have equally contributed to this work.

https://doi.org/10.1016/j.biopha.2019.108690
Received 27 December 2018; Received in revised form 12 February 2019; Accepted 14 February 2019
0753-3322/ © 2019 The Authors. Published by Elsevier Masson SAS. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/BY-NC-ND/4.0/).
Z. Ghanbari Movahed, et al.
1. Introduction
Many cancer cells exhibit persistently high reactive oxygen species
(ROS) levels as a consequence of genetic, metabolic (higher metabolism
rate) and microenvironment-associated alterations [1].
In cancer cells high ROS levels can originate from increased meta-
bolic activity, mitochondrial dysfunction, oncogene activity, peroxi-
some activity, increased activity of oxidases, cyclooxygenases, lipox-
igenases, thymidine phosphorylase and increased cellular receptor
signaling, or through crosstalk with infiltrating immune cells [2].
As reported by Spitz and colleagues cancer cells produce greater
amounts of hydrogen peroxide (H2O2) and superoxide (O2%−) compared
to normal cells [3,4].
Cancer cell mitochondria may produce greater steady-state levels of
H2O2 and O2%−, relative to normal cells [5].
Moreover, glucose deprivation, as a tumor microenvironmental
factor, induces oxidative stress in cancer cells [6].
Tumor suppressor and oncogenic pathways, which are frequently
mutated in cancer, commonly lead to increased accumulation of ROS
[7–11]. Furthermore, conditions associated with tumorigenesis such as
hypoxia, inflammation, matrix detachment and mitochondrial dys-
function can all result in excess production of ROS [12–16].
In comparison to normal cells, which are characterized by lower
intracellular ROS levels, cancer cells have a higher antioxidant capacity
to counteract and scavenge ROS which represents a selective advantage
and augments their survival under pro-oxidizing conditions, as high
antioxidant capacity impairs cellular responses to anticancer therapy
and enhances cell survival [17,18].
Studies have indicated that there is a close relationship between
cellular metabolism and redox homeostasis. Cancer cells use this con-
nection for cellular growth or to prevent oxidative damage. By other-
wise using substrates and metabolic intermediates in the biochemical
pathways which leads to the production of key antioxidant molecules,
malignant cells can directly support the mechanisms of ROS detox-
ification [14,19]. Studies have shown that acceleration of glycolysis;
i.e., the Warburg effect; and pentose phosphate pathway (PPP) in
cancer cells (human colon and breast cancer cell lines) could decrease
O2%− and H2O2 production [4] by less reliance on mitochondrial oxi-
dative phosphorylation (OXPHOS) and concomitantly augment the
redox potential via an increase in NADPH [20,21].
Warburg effect, which is a metabolic hallmark of most cancer cells,
is characterized by an excessive conversion of glucose to lactate even in
the presence of ample oxygen [22]. On the other hand, intratumoral
heterogeneity, including those involving cancer stem cells (CSCs), is
currently of interest in cancer research [23]. CSCs, characterized by the
ability to self-renew, are a small subset of cancer cells believed to be
responsible for tumor initiation, maintenance, growth, metastasis, re-
currence and drug resistance [24]. It is plausible that there exist dif-
ferences in metabolic state of heterogeneous tumors composed of dif-
ferentiated cells and CSCs. CSCs have been reported to have an
enhanced PPP metabolism [25].
Studies have shown that during oxidative stress, cancer cells will
shut down the glycolytic pathway and instead enhance glucose flux
through the PPP in order to generate more NADPH for antioxidant
defense [21].
PPP activity is increased in response to ionizing radiation, che-
motherapy or oxidative stress which increase ROS levels and trigger an
adaptive response by augmenting the PPP [26]. Similarly, the popula-
tion of CSCs increases in response to ionizing radiation, chemotherapy
or oxidative stress [27–29]. It has been documented that in multiple
cancer cell lines, the acquisition of drug resistance, a feature of CSCs, is
accompanied by enhanced oxidative PPP activity [26].
Therefore, based on the conducted studies, we hypothesize that the
metabolic deviation from glycolysis to PPP in the encounter of persis-
tent oxidative stress, might represent alterations in cancer cells towards
CSCs.
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Biomedicine & Pharmacotherapy 112 (2019) 108690
In this review, we first describe ROS and their sources as well as
control of intracellular ROS contents. In the next step we elaborate the
role of glycolysis and PPP in coping with ROS and then proceed to the
possible molecular mechanisms involved in activating the Warburg
effect and PPP in response to ROS. Finally, we discuss the possible
molecular mechanisms of metabolic deviation from glycolysis to PPP, as
an appropriate target for more effective cancer treatment.
2. Reactive oxygen species (ROS) with an emphasis on hydrogen
peroxide (H2O2)
ROS is generally taken to include the initial species produced by
oxygen reduction (hydrogen peroxide (H2O2) or superoxide (O2−%)) as
well as their secondary reactive products. Also, reactive nitrogen spe-
cies (RNS) is in common usage to describe reactive species derived from
nitric oxide (NO) [30]. H2O2, as a key metabolite in oxidative stress, is
believed to be a major player in ROS signaling owing to its physico-
chemical properties, which include long half-life, a relatively low re-
activity and the ability to diffuse through membranes [31,32]. H2O2
emerged as the main redox metabolite operative in redox sensing, sig-
naling and redox regulation in physiological conditions (1–10 nM up to
approximately 100 nM). Supraphysiological concentrations of H2O2
(> 100 nM) result in damage of biomolecules, denoted as “oxidative
distress” [33].
Intracellular ROS produced under physiologic conditions serve as
key signaling molecules regulating numerous cellular processes [34].
The abnormal accumulation of endogenous or exogenous ROS by the
mitochondrial electron transport chain (ETC) or in response to low
linear energy transfer (LET) ionizing radiation, like X-rays, may cause
protein denaturation, lipid peroxidation, or DNA mutation (oxidative
distress) [35–37].
ROS have both protumorigenic and antitumorigenic effects.
Escalated generation of ROS in tumor contributes to molecular and
biochemical alterations necessary for tumor initiation, promotion,
progression as well as chemo/radioresistance. ROS play play important
roles in tumor cell signal transduction contributing to tumor cell sur-
vival, proliferation as well as tumor angiogenesis and metastasis. For
example, PI3K/AKT/mTOR survival signaling induced by ROS, results
in the activation of mainly two signaling pathways: 1) Ras-MAPK
leading to cell proliferation and 2) PI3K-Akt-eNOS which cause meta-
bolic modulation and cell survival. ROS also inactivates phosphatase
and tensin homolog (PTEN) pathway implicated in apoptosis initiation
[38]. On the other hand, ROS can negatively regulate cellular FLICE-
inhibitory protein (c-FLIP) by augmenting its proteasomal degradation
facilitating apoptosis. Also, H2O2 can iduce dimerization of apoptosis
signal-regulating kinase 1 (ASK-1) resulting in its activation and sub-
sequent induction of apoptosis. ROS can also induce cytochrome-c (Cyt-
c) release and activate caspase-9 thereby inducing apoptosis [39].
Tumor cells are characterized by an augmented metabolic activity
leading to changes of the cellular redox state that has to cope with the
production of high levels of ROS [18]. In numerous cancer cells per-
sistently upregulated H2O2-dependent signaling pathways are im-
plicated in cell differentiation, growth and survival, yet high H2O2 le-
vels can also induce cell cycle arrest or apoptosis in cells [40]. Human
tumor cells produce high levels of H2O2 [41]. The main antioxidant
enzymes include superoxide dismutase (SOD), that converts O2−% into
H2O2, as well as catalase and glutathione peroxidase (GPX) which
convert H2O2 into water [42]. It has been shown that SOD, via its
metabolic product H2O2, can induce migration, invasion, and epithelial-
mesenchymal transition (EMT) (as a stem cell-like feature) of cancer
cells in pancreatic cancer cells through activation of the H2O2/ERK/NF-
κB axis [43].
Treatment of human malignant mesothelioma (HMM) cells with
H2O2 enhances EMT, as indicated by increased expression levels of vi-
mentin, TWIST1 and SLUG and decreased E-cadherin expression. Also,
expression of stemness genes such as oct4, sox2 and nanog significantly
Z. Ghanbari Movahed, et al.
Fig. 1. Sites for the generation of O2−% and H2O2 in mitochondria.
increases following treatment with H2O2 [44].
Therefore, regarding the importance of H2O2 in cancer biology, in
the current review we aimed to emphasize on this specific kind of ROS.
3. Sources of ROS
ROS can be found in the environment [45–48] such as in pollutants,
iron salts, radiation, and tobacco smoke or can be generated inside
[48,49] or outside the cells [50–53] which can occur through multiple
mechanisms.Generally, continuous production of O2−% and H2O2 takes
place in the mitochondria as the most important source of cellular ROS.
This is the result of the electron transport chain (ETC) located in the
mitochondrial membrane that is essential for the production of energy
within the cells (54,55). O2−% and H2O2 are produced in the mi-
tochondria as by-products of oxidative phosphorylation for ATP
synthesis and fatty acid (FA) metabolism [56,57].
In mitochondria, several proteins involved in glycolysis, β-oxida-
tion, mitochondrial electron transport, and the TCA cycle are able to
generate O2−%, H2O2 and/or other ROS (Fig. 1).
Several oxidases localized in specialized stable (peroxisomes) or
transient (endosomes, phagosomes, multivesicular bodies, etc.) single-
membrane bound organelles are capable of generating substantial
amounts of H2O2 [57].
Mammalian peroxisomes play a pivotal role in various metabolic
pathways such as fatty acid α- and β-oxidation, glyoxylate metabolism,
ether-phospholipid biosynthesis, polyamine oxidation, amino acid cat-
abolism, and the oxidative part of PPP. Interestingly, many of the en-
zymes involved in these pathways produce specific ROS or RNS as by-
products of their normal catalytic function [47].
H2O2 is the main oxidant product of xanthine oxidase (XO) [58].
Reports regarding XO-derived ROS generation frequently address XO as
the O2−%-producing form of xanthine dehydrogenase (XDH) and H2O2
is generated as a secondary byproduct of spontaneous or enzymatic
dismutation of O2−% [59].
ROS originating from cytosol could be delivered into extracellular
space (free extracellular ROS) in different ways including: secretion as
typically occurs in the case of activated degranulating leukocytes,
crossing the plasma membrane through some anion channels (O2−%)
and aquaporins (H2O2) as well as delivery via exosome release [60].
4. Control of the intracellular concentrations of ROS
Under normal physiological conditions, intracellular ROS levels are
steadily maintained to prevent cells from damage. Detoxification from
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Biomedicine & Pharmacotherapy 112 (2019) 108690
ROS is facilitated through antioxidant enzymes (i.e. SODs, peroxir-
edoxins, catalases and glutathione system) specifically scavenging dif-
ferent kinds of ROS or by non-enzymatic molecules (i.e. flavonoids,
glutathione, and vitamins A, C and E) [2]. As earlier mentioned
[4,15,19–21], several biochemical pathways play a role in ROS detox-
ification, such as glycolysis and PPP. Here we proceed to discuss these
two pathways.
4.1. Glycolysis and ROS detoxification
4.1.1. Glycolysis
Glycolysis occurs in all cells of the body. The enzymes involved in
this pathway exist in the cytosomal fraction of the cell. Glycolysis can
occur both in the absence of oxygen (anaerobic) and in the presence of
oxygen (aerobic). Under anaerobic condition, lactate is the end product
of glycolysis. In the aerobic condition, pyruvate is generated, which is
then oxidized to H2O and CO2 [61].
Two ATP molecules are synthesized in the anaerobic glycolysis
while under aerobic conditions ˜seven ATPs are synthesized from glu-
cose conversion to pyruvate [62,63].
4.1.2. Warburg effect
A key change that cancer cells have to make in order to meet their
modified energy demands is to reprogram their cellular metabolism,
one of aspects of which is to switch to an increased glycolysis pathway
in spite of having access to enough oxygen [64].
The metabolism of glucose is essential for supporting all mammalian
life. In mammals, the end product can be lactate or CO2 which is gen-
erated upon full oxidation of glucose via respiration in mitochondria. In
tumor cells and other developing or proliferating cells, glucose uptake
rate is outstandingly augmented and even in the presence of oxygen and
fully functioning mitochondria, lactate is produced. This process is re-
cognized as the Warburg effect [65].
In 1923, Otto Warburg for the first time reported an anomalous
characteristic of energy metabolism in cancer cells [66,67]. Warburg
observed that cancer cells, even under fully oxygenated conditions,
were characterized by accelerated glycolysis and excessive lactate for-
mation [68] Later in 1972, his discovery was named the ‘Warburg Ef-
fect’ by Efraim Racker [69].
Respiration, generating some 32/30 mol of ATP per mole of glucose
[63,70], is downregulated in cancer cells in favor of fermentation which
only produces two moles of ATP per mole of glucose. Therefore, to
supply sufficient ATP, the Warburg effect is associated with an in-
creased uptake of glucose (Fig. 2). Using this low ATP-yielding pathway
Z. Ghanbari Movahed, et al.
Fig. 2. Difference in glucose metabolic pathways in the presence and absence of oxygen in normal vs cancer cells.
by cancer cells seems amazing and several advantages of the Warburg
effect for cancer cells have been proposed [71].
Moreover, glycolysis upregulation is not only for ATP synthesis but
also for synthesis of biomass including ribonucleotides [72] and amino
acids [70] for proliferation and growth in an ever-changing micro-
environment under several material limitations like shortages of oxygen
and nutrients. Upregulation of glycolysis could also be considered as a
way to reduce ROS and therefore oxidative stress in cancer cells
[73,74].
4.1.3. Warburg effect: coping with ROS
In most mammalian cells, mitochondria are an important source of
H2O2 and O2−% [53]. Upregulation of glycolysis rate in cancer cell
energy metabolism could decrease the production of H2O2 and O2−% by
less reliance on mitochondrial OXPHOS (Fig. 3) [75]
4.2. PPP and ROS detoxification
4.2.1. Pentose phosphate pathway (PPP)
PPP is classified into two biochemical branches known as the oxi-
dative and non-oxidative PPP. Non-oxidative PPP reactions, that occur
virtually ubiquitously, play a central metabolic role by providing the
RNA backbone precursors erythrose 4-phosphate and ribose 5-phos-
phate as precursors for aromatic amino acids. On the contrary, oxida-
tive branch reactions of PPP are not universal and are absent in many
aerobic and thermophilic organisms [76–78]. However, the oxidative
branch, being highly active in most of the eukaryotes, via the successive
reactions of G6PD, 6-phosphogluconolactonase (6PGL) and 6-phos-
phogluconate dehydrogenase (6PGDH) converts the glycolytic meta-
bolite glucose 6-phosphate (G6P) into ribulose 5-phosphate. This me-
tabolic sequence produces two NADPH molecules per each glucose 6-
phosphate metabolized [79].
4.2.2. PPP: coping with ROS
NADPH, in its reduced form, is the donor of reductive potential to
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Biomedicine & Pharmacotherapy 112 (2019) 108690
thioredoxins and glutathione which in turn are used (directly or in-
directly) by glutaredoxins, peroxiredoxins and glutathione peroxidases
to neutralize ROS. Therefore, for a large number of ROS-detoxifying
enzymes, NADPH acts as the ultimate donor of reductive power [79].
Several metabolic pathways such as the reactions catalyzed by the
malic enzymes (ME), folate dehydrogenases (FDH) and isocitrate de-
hydrogenases (IDH) can generate NADPH; but the main sources of
cellular NADPH are two enzymes of the oxidative branch of PPP: the
PPP rate-limiting enzyme G6PD and 6PGDH [80].
4.3. Mechanisms of activating the Warburg effect in response to ROS
(DIRECT)
4.3.1. GLUT1 and ROS
In order to involve in the Warburg effect, the cancer cells utilize
much more glucose compared to normal cells which derives most of
their energy from aerobic metabolism [81]. Accordingly the ability of
such cells to bring glucose into their cytoplasm needs to be increased.
Various glucose transporters (GLUTs), primarily GLUTs 1, 3, and 4,
have been shown to be upregulated in endometrial cancer, ovarian
cancer, gastric cancer, meningiomas, glioblastomas and squamous cell
carcinomas as well as in many of these cell lines [82]. GLUT-1, an
important member of GLUT family, is a basic high-affinity glucose
transporter normally expressed in erythrocytes, renal tubules, placenta
and endothelial cells. GLUT-1 expression has been shown to be asso-
ciated with tumor aggressiveness and poor prognosis in many cancers
including lung, breast, stomach and kidney [83].
Various signaling pathways are involved in glucose uptake stimu-
lation as an adaptive response during oxidative stress. ROS-induced
signals, by upregulating GLUT protein expression, induce glucose up-
take. The rapid induction of glucose uptake can occur at the level of
GLUT translocation and regulation of GLUT intrinsic activity.
Interestingly, a number of the signaling proteins are ROS-sensitive
(such as AMP-activated protein kinase (AMPK) and thioredoxin-inter-
acting protein (TXNIP)). This indicates that glucose uptake stimulation
Z. Ghanbari Movahed, et al.
Fig. 3. Warburg effect reduces the production of ROS by less reliance on mitochondrial OXPHOS. Pyruvate is a source for OXPHOS. Mitochondrial OXPHOS is a main
source of ROS production. By increasing the conversion of pyruvate to lactate, Warburg effect reduces ROS production by the mitochondrial OXPHOS.
induced by ROS is part of a/an (adaptive) mechanism triggered by
oxidative and/or metabolic stress [54]. Translocation of GLUT1 to the
plasma membrane is regulated by ataxia telangiectasia mutated (ATM)
protein [84]. This protein is a member of the phosphatidylinositol-3-
kinase-related protein kinases (PIKKs) family playing an important role
in the response to DNA damage after which it becomes phosphorylated.
However, ATM can also localize to the cytosol where following acti-
vation by ROS, it is involved in cytosolic signaling [85]. ATM, following
thiol oxidation by ROS, forms an active dimer of two covalently linked
monomers. Activated ATM localizes near the mitochondria which are
necessary for ROS-induced ATM activation. This suggests that mi-
tochondrial ROS stimulate ATM dimerization and activation. There are
evidence regarding the activation of ATM by ROS during treatment
with the chemotherapeutic agent doxorubicin [54].
4.4. Mechanisms of activating the Warburg effect in response to ROS
(INDIRECT)
4.4.1. Hypoxia-inducible factor 1 (HIF-1) and ROS
4.4.1.1. HIF-1. Hypoxia-inducible factor 1 (HIF-1) is a member of a
family of heterodimeric transcription factors which contain a bHLH-
PAS (basic helix-loop-helix/Per-Arnt-Sim) domain. HIF-1, with a
ubiquitous expression, is the key player in regulating hypoxic genes
and mediates the upregulation of hypoxia-inducible genes [86,87]. HIF-
1 comprises an α-subunit isoform (HIF-1 α, HIF-2 α or HIF-3 α) and a
common β-subunit (HIF-1 β) identical with aryl-hydrocarbon receptor
nuclear translocator (ARNT). Both of these subunits are constitutively
expressed in the cell with the α-subunit being oxygen-labile while the β-
subunit is not sensitive to changes in oxygenation [88]. Target genes of
HIF-1 are implicated in angiogenesis (VEGF or vascular endothelial
growth factor), glucose metabolism and glucose transport (GLUT-1,
glycolytic enzymes), iron transport (transferrin, transferrin receptor)
and hematopoiesis (erythropoietin) and aim for the adaptation to
hypoxia on systemic, local and cellular levels [89]. More than 100
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Biomedicine & Pharmacotherapy 112 (2019) 108690
genes with different functions have been identified as the target genes
activated by HIF-1 [90]. On the other hand, HIF-1α activity and
accumulation are themselves found to be regulated by different
cellular levels of many other factors including Hsp90, TSGA10, etc.
[91,92].
4.4.2. HIF-1 regulates glycolysis and the Warburg effect
GLUT1 and GLUT3 are needed for glucose shuttling across cell
membranes. Due to the high affinity of GLUT1 and GLUT3 for glucose,
expression of these two molecules guarantees efficient glucose uptake
even in the conditions of glucose shortage. Both of the two isoforms
have a HIF-1-inducible gene expression coupling the glycolytic switch
to increased uptake of glucose in hypoxic cancer cells [93].
Hexokinase (HK) 2 that catalyses the first rate-limiting reaction of
glycolysis (glucose phosphorylation to G6P) involving phosphate
transfer from ATP, is a member of the HK family of enzymes. The ne-
gatively charged G6P is trapped inside the cell and can fuel both gly-
colysis and PPP. In comparison with other isoforms of HK, HK2 is a HIF-
1 target gene product [94].
Fructose-2,6-bisphosphate (F2,6-BP) is a key glycolysis regulator
which serves as an allosteric activator of phosphofructokinase 1 (PFK1),
one of the rate-controlling enzymes of glycolysis. F2,6-BP is generated
from fructose-6-phosphate (F6P) by the action of a family of homo-
dimeric enzymes known as 6-phosphofructo-2-kinase/fructose-2,6-bi-
sphosphatases (PFKFB). The family consists of four members among
which PFKFB1, PFKFB2, and PFKFB4 exhibit equal PFK2 (ATP-depen-
dent phosphorylation of F6P to F2,6-BP) and FBPase (depho-
sphorylation of F2,6-BP to F6P) activities under basal conditions, while
PFKFB3 has high PFK2 and almost no FBPase activity [94,95,96]. Hy-
poxia can induce the transcription of all four PFKFB genes but the main
induction is seen for the PFKFB3 gene which is a target of HIF-1
[97,98]. PFKFB3 that is highly expressed in several tumor types, sus-
tains high-rate glycolysis [93].
Pyruvate dehydrogenase (PDH), the enzyme which commits
Z. Ghanbari Movahed, et al.
pyruvate to enter into the Krebs cycle, is inhibited by phosphorylation
via PDK-1 (pyruvate dehydrogenase kinase 1). Under hypoxic condi-
tions, PDH inhibition follows two main objectives: 1) orienting pyr-
uvate to the lactate dehydrogenase (LDH) 5 reaction to produce NAD+
and 2) preventing the excessive production of ROS by uncoupled mi-
tochondria [99,100]. HIF-1α also hinders mitochondrial respiration
through inducing PDK-1 which is a PDH inhibitor [99]. High expression
of PDK-1 strongly correlates with poor outcome in head-and-neck
cancer [101].
LDHs are comprised of up to four copies of two different subunits:
subunit LDH-H, ubiquitously expressed in healthy tissues, is encoded by
the ldhb gene, while subunit LDH-M is encoded by the HIF-1-target gene
ldha and is hence induced by hypoxia. In comparison with LDH-H, LDH-
M has a higher Km for pyruvate and a higher Vmax for pyruvate re-
duction [101,102]. As a result, LDH5/LDH-4 M preferentially catalyzes
the reduction of pyruvate into lactate and plays pivotal roles in re-
sistance to apoptosis and in the maintenance of a high glycolytic flux.
Augmented expression of LDH5 has an unfavorable prognostic sig-
nificance for many human tumors [103–105]. Conversely, silencing of
LDH1/LDH-4H is most commonly observed in glycolytic cancer cells, a
process which involves hypermethylation of the ldhb gene promoter
[106,107].
Pyruvate kinase (PK) is one of the rate-limiting enzymes of the
glycolysis pathway which catalyzes the irreversible transpho-
sphorylation between phosphoenolpyruvate (PEP) and adenosine di-
phosphate (ADP) producing pyruvate and ATP. This reaction is a
committed step which results in glycolysis or oxidative phosphorylation
of pyruvate. In mammals, the PK family comprises of four isoforms: red
blood cell PK (PKR); liver-type PK (PKL); and PK muscle isozyme M1
and M2 (PKM1 and PKM2, respectively). PKM1 and PKM2 are both
generated via alternative splicing of the primary RNA transcript of the
pkm gene. The constitutively active PKM1 is the non-allosteric isoform,
expressed in terminally differentiated tissues requiring a large ATP
supply. By contrast, PKM2, which is subject to complex allosteric reg-
ulation, is expressed in proliferating cells and cancer cells which have
anabolic functions [108]. HIF-1 activates pyruvate kinase M2 (PKM2)
expression and on the other hand, PKM2 functions as a transcriptional
coactivator for HIF-1α. HIF-1 activates PKM2 transcription leading to
the production of a protein which interacts with HIF-1α to stimulate
coactivator recruitment, chromatin binding, and transcriptional acti-
vation. This feed-forward loop augments expression of HIF-1 target
genes which code for proteins mediating the metabolic reprogramming
of cancer cells (e.g., LDH-M and PDK1) [109]. Sun et al showed that
PKM2 is a key glycolytic enzyme in the oncogenic mTOR-induced
Warbug effect, in which c-Myc-hnRNP and HIF-1α cascades act as the
transducers of mTOR regulation of PKM2 [108].
4.4.3. ROS regulates HIF-1
HIF-1 plays a key role in the cellular adaptive response to hypoxia,
which is closely related to pathophysiological conditions, including
cancer [110]. ROS induced by a direct application of H2O2 (0.5 mM)
and menadione (100 μM) to the human prostate carcinoma cell line,
DU145, leads to HIF-1α protein accumulation through attenuating its
degradation and activating its transcriptional activity in an AMPK-de-
pendent manner. Inhibition of AMPK increases HIF-1α ubiquitination
under oxidative stress conditions. Also, HIF-1 regulation by AMPK in
response to H2O2 is under the control of Janus kinase 2 and c-Jun N-
terminal kinase pathways. Collectively, AMPK can be considered as a
pivotal determinant of HIF-1 functions in response to H2O2 and might
play a role in the sophisticated HIF-1 regulatory mechanisms (Fig. 4)
[111].
Hepatoma cells have higher free radicals levels compared to im-
mortalized normal hepatocyte cells. In hepatoma cells, increased level
of ROS stress (SOD and XO gene transfection were used to produce
various cellular redox levels) can directly upregulate HIF-1 and induce
glycolysis without requiring a hypoxic condition. This explains the
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Biomedicine & Pharmacotherapy 112 (2019) 108690
mechanism by which aerobic glycolysis, i.e. the Warburg effect arises
and it is suggested that the Warburg effect is related to an inherently
high cellular ROS and HIF-1 levels [112].
4.5. Mechanisms of activating PPP in response to ROS
In response to ROS, cancer cells might initially upregulate glycolysis
and then following persistent ROS exposure the metabolic balance is
skewed from glycolysis pathway to PPP to generate more NADPH for
antioxidant defense.
PK is a regulator for cellular anti-oxidative metabolism. PKM2 in-
hibition by ROS contributes to cellular antioxidant responses [73].
However, this protein plays a unique regulatory role so that its de-
creased catalytic activity is associated with tumor progression
[113,114]. When the kinase activity of PK is low — as in cancer cells—
its substrate, phosphoenol pyruvate, accumulates which subsequently
inhibits the glycolytic enzyme triose phosphate isomerase (TPI) re-
sulting in the activation of a pathway alternative to glycolysis i.e., PPP
[115,116]. Augmented PPP activity protects the cells against ROS in at
least two ways. First, it provides NADPH, a reducing molecule required
for antioxidant enzymes activity and for the recycling of the antioxidant
peptide glutathione (GSH). Also, NADPH compensates for the redox
imbalance caused by increased synthesis of nucleotides and fatty-acids
[117]. Second, PPP regulates gene expression in favor of adaptation to
oxidative stress [118]. Several types of oxidative stresses, such as H2O2,
hypoxia and diamide inactivate PKM2 in human cancer cells. Anasta-
siou D et al. showed that in A549 human lung cancer cells exposure to
H2O2 (1 mM), diamide (250 μM), as a thiol-oxidizing compound, or
hypoxia (1% O2) [which causes increased ROS generation by mi-
tochondrial complex III [119]] was associated with increased in-
tracellular concentrations of ROS leading to decreased PKM2 activity.
In that study, acute increases of intracellular ROS levels in human lung
cancer cells caused inhibition of the glycolytic enzyme PKM2 through
oxidation of Cys358. This inhibition of PKM2 is required to divert
glucose flux into PPP thereby generating sufficient reducing potential
for detoxification of ROS [73].
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is an
evolutionarily conserved enzyme, controls glucose flux through the
canonical Embden-Meyerhof glycolytic pathway. GAPDH, during pro-
oxidant conditions, contributes to a glycolytic bottleneck favoring a
metabolic switch towards PPP [120].
GAPDH inhibition helps divert glycolytic flux into the oxidative
phase of PPP by allowing metabolites to accumulate upstream of the
inhibition point consistent with the observed induction of PPP enzymes
following H2O2 treatment [121,122].
In mammalian cells (P388D1 cells), GAPDH can be inhibited within
minutes of exposure to H2O2 (100 μM) mainly via direct inactivation of
the enzyme and loss of NAD + cofactor likely through PARP (Poly
(ADP-ribose) polymerase) activation [123,124]. The active site cysteine
of GAPDH is highly sensitive to inhibitory oxidative modifications of
RNS and ROS [125].
TIGAR, or TP53-induced glycolysis and apoptosis regulator, is
identified as a P53 target gene induced by ionizing radiation. TIGAR
has a single bisphosphatase (BPase) activity which degrades F-2,6-BP to
F-6-P. TIGAR-mediated decrease in F-2,6-BP levels, inhibits glycolytic
flux downstream of PFK1. PFK1 inhibition results in the accumulation
of G6P and F6P pools as their consumption is greatly diminished. The
increased G6P can flow into the oxidative arm of the PPP to produce
NADPH. UV and ROS stress can induce P53-dependent TIGAR activa-
tion which inhibits PFK1 [125].
5. In response to increased ROS, which one do the cancer cells
increase? Glycolysis or PPP
Studies have also shown that increased glycolysis and PPP in cancer
cells could reduce the production of ROS [20,21].
Z. Ghanbari Movahed, et al.
Activation of oxidative PPP, through increasing NADPH production
and thus enhanced intracellular redox power of cancer cells, can help
transformed cells escape oxidative stress [126]. The increased PPP in
cancer cells produces high levels of NADPH to reduce ROS while con-
comitantly producing high nucleotide levels for DNA synthesis and
repair. These activities of PPP might induce resistance to certain cancer
therapies which enhance oxidative stress or DNA damage [26].
As demonstrated by several studies, alterations of PPP considerably
contribute to tumor growth and survival under certain stress condi-
tions. During oxidative stress, cancer cells are able to cope with oxi-
dative stress by redirecting the metabolic flux from glycolysis to PPP
[21,127].
Therefore, cancer cells might initially respond to ROS by increasing
glycolysis rate and then in response to increased ROS level/exposure,
they increase PPP.
6. Glycolysis deviation to PPP: from cancer cell to cancer stem cell
Cancer stem cells (CSCs) are a small subset of cancer cells capable of
self-renewing and initiating tumor growth. These cells are implicated in
cancer initiation, maintenance, metastasis and recurrence (29).
Traditional therapies of cancer such as chemo- and radiotherapy have
several limitations that result in treatment failure and cancer recur-
rence.
The Warburg effect is thought to be a common phenomenon in all
tumor cells. However, there seems to exist differences regarding me-
tabolic state in heterogeneous tumors composed of differentiated cells
and CSCs. There are contradictory literature data regarding the meta-
bolic states of CSCs and their differentiated progeny. According to some
studies comparing the metabolic state of leukemia, glioma, and breast
CSCs with the differentiated cell population, CSCs are more dependent
on oxidative phosphorylation than differentiated cells [128–130], while
some other studies point to more dependence of these cells on anae-
robic glucose metabolism [131–133].
CD44 expression by CSCs could be considered as one of the evidence
showing the metabolic deviation from glycolysis to PPP in such cells.
CD44, as one of CSC markers of various kinds of solid tumors, is cate-
gorized into standard (without variables exons, or CD44 s) and variant
(with variable exons) isoforms. Intracellular domain of CD44 s has been
shown to interact with and suppress PKM2 through increasing its
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Biomedicine & Pharmacotherapy 112 (2019) 108690
Fig. 4. HIF-1 mediated effects of ROS (H2O2) on upregulation of
glycolysis. A number of ROS can activate HIF-1α. Activated HIF-
1α upregulates glucose transporters GLUT1 and GLUT3 to in-
crease glucose uptake into cancer cells. HIF-1α accelerates gly-
colysis by upregulating ENO, ALD, PGK1 and PKM2 and meta-
bolizing pyruvate. Pyruvate is not converted to acetyl CoA
because HIF-1α upregulates PDK-1, which inhibits PDH.
phosphorylation resulting in antioxidant status (increased GSH and
reduced ROS) of CSCs. Low PKM2 activity, by inhibiting the conversion
of pyruvate to lactate, promotes the flow of glycolytic intermediates
into PPP. Also, cancer stem-like cells expressing high levels of CD44
variant 8–10 (CD44v) do not show accumulation of intracellular ROS
and are protected from redox stress. CD44v stabilizes cysteine/gluta-
mate antiporter at the cellular membrane which promotes the synthesis
of GSH. On the other hand, cancer cells with high PKM2 level/activity
use a c-Myc-mediated strategy to preferentially splice the M2 isoform
over M1. It has been found that cells with high c-Myc activity also show
high PKM2/PKM1 ratios. A great deal of evidence indicates that c-Myc
stimulates glycolysis and is required to coordinate with HIF-1 for the
regulation of cellular responses to hypoxia. Therefore, high levels of c-
Myc activity are responsible for the increased PKM2/PKM1 ratios in
cancer cells. C-Myc, in combination with HIFs, can promote glycolysis
by upregulating GLUT1, HK2 and PDK1. It has been shown that in
gastric cancer cells, CD44v and c-Myc exhibit an inversed expression
manner. FBW7, a ubiquitin ligase component, can mediate ubiquitin-
proteasome-dependent degradation of c-Myc. Tumor tissue at the in-
vasive front is regarded to be composed of heterogeneous cellular po-
pulations including proliferative cancer stem-like cells (CD44vhigh,
FBW7low, c-Mychigh) and dormant cancer stem-like cells (CD44vhigh,
FBW7high, c-Myclow). Accordingly, CSCs with high expression of CD44v
and FBW7 and low expression of c-Myc tend to be quiescent (G0/G1
dormant phase) [134–136].
It has been determined that histone deacetylase (HDAC) inhibitors,
by reprogramming differentiated cancer cells into stem-like cells, pro-
mote CSC expansion. In a study, Debeb et al. investigated metabolic
differences between stem-like breast cancer cells and differentiated
cancer cells. According to the results of their study, HDAC inhibitor-
induced stem-like cancer cells exhibit an enhanced PPP metabolism.
PPP plays a substantial role in the production of cellular NADPH which
is required for fatty acids synthesis and intracellular ROS detoxification.
As expected, aldehyde dehydrogenase− (ALDH−) or ALDH + cells
treated with valproic acid (VA) or suberoylanilide hydroxamic acid
(SAHA) (as HDAC inhibitors), had significantly higher levels of NADPH
compared to the vehicle-treated cells [25]. Also in a study conducted by
La Noce et al., HDAC2 depletion was shown to promote stemness in
osteosarcoma cell lines. They investigated the effects of valproic acid
(VPA), an HDAC inhibitor, and 5′azacytidine (DAC), a demethylating
Z. Ghanbari Movahed, et al.
agent, on the stem phenotype of two osteosarcoma cell lines MG63 and
Saos2. They found an increased expression of the stemness markers:
CD133, OCT4, SOX2 and NANOG as well as increased sarcospheres and
colonies formation in soft agar (as a tumor stemness characteristic)
following treatment with VPA and DAC alone or their combination.
Also, they performed an analysis of histone modifications previously
shown to be implicated in embryonic stemness and interestingly found
that treatments with VPA and DAC alone and in combination, induced a
decrease of H3K9me3 (trimethylation of histone 3 lysin 9) and
H3K27me3 (as repressive histone markers) and an increase of
H3K4me2 and H3K4me3 (as active histone markers) that were asso-
ciated with an increased acetylation of histone H3 and a significant
reduction of global DNA methylation compared to untreated cells.
Furthermore. HDAC2-silenced osteosarcoma cell lines acquired a stem
phenotype and promoted in vivo tumorigenesis in mouse xenografts.
Overall, they showed that VPA and DAC induce expansion of osteo-
sarcoma CSCs and that HDAC2 is a key mediator in regulating both CSC
phenotype and cancer growth in vivo [137]. In addition, PPP activity is
enhanced in response to oxidative stress [138], ionizing radiation [139]
or chemotherapies [140], which induce high levels of ROS and trigger
an adaptive response by augmenting PPP. In multiple cancer cell lines,
acquisition of drug resistance has been documented to be associated
with elevated oxidative PPP activities. Sustained high levels of GSH and
G6PD are hallmarks of enhanced oxidative PPP activity accounting for
drug resistance [141,142]. Therefore, PPP inhibition seems to be a
promising strategy for cancer targeting. Mele and coleagues in 2018
showed anti-PPP effects of the natural molecule polydatin (3,4′,5-tri-
hydroxystilbene-3-β-D-glucoside; transresveratrol 3-β-mono-D-gluco-
side; piceid) through targeting G6PD. In their study, blockade by
polydatin of G6PD was shown to be associated with ROS accumulation
and strong increase of endoplasmic reticulum (ER) stress leading to cell
cycle arrest at S phase, increased apoptosis (50%), and inhibition of in
vitro invasion (60%). Polydatin inhibited proliferation of head and neck
squamous cell carcinoma (HNSCC) cell lines and caused ER-induced cell
death (as a result of ROS accumulation) due to collaborating actions of
inositol-requiring enzyme 1 (IRE1; an enzyme activated during ER
stress with intrinsic kinase and endoribonuclease activities) and protein
kinase RNA-like ER kinase (PERK) in inducing the transcription of
CHOP (CCAAT-enhancer-binding protein homologous protein) which is
involved in apoptosis activation. G6PD overexpression was shown to
counteract polydatin antitumor effects. In an experimental orthotopic
metastatic model of tongue cancer, polydatin reduced tumor size and
inhibited lymph node metastases. Also, polydatin increased che-
motherapeutic effects of cisplatin and afatinib [143].
Studies have also shown that exposure to stress associates with an
increase in stem cells populations, for example: Ionizing radiation (IR)
can increase ROS production [144]. Radiolysis of water and early ac-
tivation of nitric oxide synthases are main sources of ROS and RNS in
irradiated cells under ambient oxygen. Interestingly, the yield of these
species is highly modulated by different radiation types. Increasing LET
of the irradiating particles, is associated with an increased yield of
molecular products (such as H2O2) and a corresponding decreased yield
of radicals (such as %OH). In contrast, O2%− (or HO2%) is the most
abundant radical species generated by radiations with high LET char-
acter [145,146].
ROS have been demonstrated to have a significant role in mediating
the biological effects of IR. Although considered as a standard therapy
for a variety of malignant tumors, IR also paradoxically promotes tumor
metastasis and recurrence [144]. The epithelial mesenchymal transition
(EMT) has been demonstrated to provide cancer cells with migratory
and invasive characteristics, enabling the metastasis initiation
[147–149]. It is known that IR induces EMT. EMT might be closely
linked to CSCs and the metabolic reprogramming of cancer cells. IR has
been demonstrated to induce CSC phenotype in multiple cancers, such
as: lung, breast and prostate cancers, as well as melanoma. Genotoxic
stress owing to chemotherapy or IR promotes a CSC-like phenotype by
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Biomedicine & Pharmacotherapy 112 (2019) 108690
increasing the production of ROS [144]. It has been shown that IR can
induce reprogramming of differentiated cancer cells into CSCs [150]. In
patients with prostate cancer, radiotherapy elevates the CD44+ cells
population that show CSC properties [151]. Also, IR induces the re-
expression of stem cell regulators like OCT4, SOX2, NANOG, and KLF4,
to promote stemness in cancer cells [144].
Induced breast cancer stem cells (iBCSCs) have been found to be
generated by radiation. Dr. Pajonk and colleagues, after eliminating the
smaller pool of BCSCs and then irradiating the remaining breast cancer
cells, put these cells in mice. They found that the ability of these iBCSCs
to form tumors increased more than 30 folds compared to non-irra-
diated breast cancer cells. Their findings demonstrate that threatening
differentiated breast cancer cells by exerting challenges on them (such
as radiation), induces the generation of iBCSCs which might, along with
surviving CSCs, produce more tumors [150].
Normal stem cells have been shown to contain lower ROS levels
compared to their more mature progeny, a characteristic which is cri-
tical for the maintenance of stem cell function. Low ROS levels are
required to maintain key properties of stem cells including quiescence
and self-renewal whereas high ROS levels result in stem cell differ-
entiation, senescence and apoptosis. Therefore, ROS production in stem
cells is tightly regulated to ensure the ability to maintain tissue
homeostasis highlighting the presence of enhanced ROS scavenging
systems in such cells. Similar to normal tissue stem cells, CSCs exhibit
lower intracellular ROS contents and enhanced ROS defense (anti-
oxidant capabilities) in comparison with their non-tumorigenic progeny
which may contribute to tumor chemo/radio-resistance [151,5]. Nu-
clear factor erythroid2-related factor-2 (NRF2) is a central and pivotal
transcription factor involved in cellular antioxidant defense system
(genes encoding G6PD and 6PGD enzymes of oxidative PPP as the most
important providers of cellular NAPH responsible for antioxidant de-
fense, are major targets of this transcription factor) and thus plays a key
role in cell homeostasis, proliferation and chemo/radioresistance. In
various cancers, inhibition of NRF2-dependent antioxidant response
using siRNA can lead to increased sensitivity to 5-FU, cisplatin, gem-
citabine and doxorubicin. Considered as a potential mediator of cancer
cell resistance, NRF-2 has been shown to be associated with cancer cell
stemness. For example, its silencing reduces the expression of aldehyde
dehydrogenase family, member A1 (ALDH1A1) and ALDH3A1 as bio-
markers of pancreatic cancer stemness [152]. Taking these in mind and
also regarding the fact that the major part of cellular NADPH (re-
sponsible for higher antioxidant capabilities and drug resistance) is
produced in PPP, high PPP rate is deduced to play a key role in the
function as well as survival of CSCs. Indeed, although augmented
aerobic glycolysis (Warburg effect) and function in parallel to enhanced
PPP (which is necessary for providing nucleotides as building blocks for
proliferation as well as supports high antioxidant capabilities in cancer
cells) to support tumor growth [21], the key metabolic balance devia-
tion from glycolysis to PPP and more enhanced oxidative PPP rate
would occur in the encounter of increased ROS level/exposure and
seem indispensable for development of CSC.
7. Dual effects of ROS on HIF-1
Increased levels of ROS in cancer cells can activate glycolysis, either
directly or indirectly. As previously mentioned, ROS can directly in-
fluence on glucose transporters and indirectly activate glycolysis
through affecting the HIF-1 axis.
O2−% and H2O2 are two species initiating cascades of redox chem-
istry which are central to redox biology. A network consisting of redox
active small molecules and enzymes maintains low steady-state levels of
O2%− and H2O2. Different redox couples sense changes in O2−% and
H2O2 flux, each resulting in distinct patterns of gene expression. H2O2,
as a signaling molecule, is a two-electron oxidant ideal for altering the
redox poise of thiols, thereby activating signaling pathways that can
result in quiescence or differentiation, or at very high flux, apoptosis
Z. Ghanbari Movahed, et al.
and cell death. In contrast, a subset of iron-containing enzymes is the
primary target of O2−%. Changes in the activity of these enzymes fol-
lowing reaction with O2−% result in changes in metabolism and gene
expression patterns which promote cell division. Examples are activa-
tion of the HIF system via prolyl hydroxylases and the inactivation of
aconitase. Superoxide dismutase, at the fulcrum of these two signaling
pathways, contributes to the balance between 1-electron and 2-electron
signaling. Therefore, the traditional view of antioxidant enzymes needs
to be expanded. The antioxidant network of small molecules, proteins,
and enzymes, that senses changes in ROS/RNS flux, creates the ap-
propriate balance between 1-electron and 2-electron signaling. As a
result, this apparent antioxidant network is best viewed as a set of in-
terconnected redox networks establishing the basic biology of cells,
tissues, and organisms [153].
Several reports suggest that mitochondria-derived ROS are required
for induction of HIF-1 during hypoxia, the effect that is mediated by
H2O2. Exogenous H2O2 has also been observed to stabilize HIF-1 (in
Hep3B cells treated with 25 or 40 μM H2O2) and catalase over-
expression prevents HIF-1 activity [154]. The PI3K and MAPK path-
ways might mediate H2O2 -induced HIF-1 stabilization, as these path-
ways are known to stabilize HIF-1 and H2O2 activates these pathways
[155–157].
Since H2O2 and HIF-1 levels are thought to be increased in cancer
cells and H2O2 is known to stabilize HIF-1, it seems conceivable that
increased HIF-1 levels in cancer cells might be due to elevated gen-
eration of H2O2 by these cells [154].
Activation of HIF not only elevates anaerobic glycolysis, but also
attenuates mitochondrial respiration. The former occurs by upregula-
tion of the genes that encode GLUT, glucokinases, aldolase A and LDH-
M, the enzymes involved in the conversion of pyruvate to lactate. The
latter occurs through the induction of PDK1, which inhibits PDH in-
volved in the conversion of pyruvate to acetyl-CoA in the mitochondria.
These two phenomena are known to hinder the entry of pyruvate to the
Krebs cycle and divert pyruvate toward lactate formation through
glycolysis [158].
Studies show that MnSOD regulates HIF-1α expression through
modulating the steady-state level of O2−%. In MCF-7 cells an siRNA
meadiated decrease of MnSOD was shown to be associated with an
increase in the hypoxic accumulation of HIF-1α and removal of O2−%
using O2−% scavenger Tempol or an SOD mimic (AEOL10113) or spin
traps (α-4-pyridyl-1-oxide-N-tert-butylnitrone and 5,5-dimethyl-1-pyr-
roline N-oxide) led to a decrease in HIF-1α protein, consistent with the
hypothesis that O2−% acts as an important molecular effecter in hypoxic
stabilization of HIF-1α. The evidence demonstrated that O2−% can
contribute to the stabilization of HIF-1α [159].
Paradoxically, accumulated evidence demonstrates that cellular
H2O2 are able to downregulate the stabilization of HIF-1α in cancer
cells. For instance, exposure of Hela cells to H2O2 (1 mM) selectively
inhibits the accumulation of HIF-1α protein in hypoxia [87]. In HepG2
cells, inhibition of the Fenton reaction increases the levels of HIF-1α,
HIF-1 transactivation, and also activates expression of the HIF-1 target
genes [160]. According to other investigations, increased ROS levels in
different cancer cells, also impairs the hypoxic signaling mechanism
and HIF-1α expression [161]. In addition, in human breast carcinoma
MCF-7 cells, over-expression of thioredoxin, a thiol reducing protein,
potentiates hypoxia-induced gene activation [162]. In normoxic cells,
the dramatic effect observed with the reducing thiol agent N-mercap-
topropionylglycine on erythropoietin (EPO) gene activation and HIF-1α
formation provides a strong indication regarding the ability of ROS to
downregulate HIF-1α in Hep 3B and B-1 cells [163]. The mechanisms
through which ROS destabilizes HIF-1α may involve: 1- increasing HIF-
prolyl hydroxylase activity; 2- activating prolyl hydroxylase (via acting
like oxygen); and 3- increasing oxidation of HIF-1α protein, as mildly
oxidized proteins more likely can be degraded by ubiquitin-proteasome
systems [161].
On the other hand, it has been shown that ROS may promote HIF-1α
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Biomedicine & Pharmacotherapy 112 (2019) 108690
degradation and reduce HIF-1α levels in ischemic neurons and brains.
Ethyl 3,4-dihydroxybenzoic acid (DHB) and dimethyloxallyl glycine
(DMOG) are prolyl hydroxylase inhibitors and well-known activators of
HIF-1. Recently, treatment with either DMOG or DHB during nerve
growth factor (NGF) deprivation has been observed to suppress the
accumulation of ROS to levels that are only slightly above the basal
level measured in NGF-maintained neurons [164].
These results imply that inhibiting ROS might play a role in DHB
and DMOG ability to increase HIF-1 expression. In explants of human
pial arteries, superoxide anion radical produced by xanthine/xanthine
oxidase decrease HIF-1α levels [165]. Co-expression of the redox factor-
1 or the antioxidant pyrrolidine dithiocarbamate increase HIF-1α levels
in RacG12V-cells [166]. Glucose is an essential substrate required for
production of the main intracellular reductants GSH and NADPH by
PPP and subsequently sustains a reduced cellular milieu in the brain. As
a consequence, glucose deprivation would lead to a condition of me-
tabolic oxidative stress. Indeed, glucose deprivation has been reported
to increase ROS production in neurons under hypoxic conditions. A
negative relationship has been observed between HIF-1α and H2O2
levels. Moreover, N-acetyl cysteine (a ROS scavenger) elevates protein
level of HIF-1α whereas L-buthionine sulfoximine, which induces oxi-
dative stress, inhibits HIF-1α expression in hypoxic neurons [161]. This
finding suggests that ROS does not promote HIF-1α protein stabiliza-
tion, but rather induces its degradation in hypoxic neurons.
Though, it is not obvious how ROS induces HIF-1α degradation in
ischemic brain, ROS might promote HIF-1α degradation through two
mechanisms. First: activation of proteasomal degradation pathways.
The proteasomal proteolytic pathways are considered as the main me-
chanisms through which many intracellular proteins are degraded
[167]. These pathways play pivotal roles in the regulation of key cel-
lular processes including apoptosis, differentiation, cytokine-induced
gene expression, and the stress response [168–170]. There are two
forms of proteasome complexes: 26S and 20S. Protein substrate speci-
ficities are usually different between the two complexes. The 26S pro-
teasomal pathway is ubiquitin-dependent and is a key pathway for the
degradation of proteins in cells while the 20S proteasomal pathway is
ubiquitin-independent and is primarily used for the degradation of
cellular oxidized proteins in oxidative stress conditions [167,171–174].
Under hypoxic conditions, ROS may activate the 26S pathway through
increasing prolyl hydroxylase activity [175–178]. ROS may also in-
crease 20S proteasomal activity [179]. HIF-1α, with its cysteine and
methionine residues being vulnerable to oxidative damages, is sensitive
to oxidative attacks [180,181]. In fact, according to a recent publica-
tion, the 20S degradation pathway appears to be implicated in the
turnover of HIF-1α under hypoxic conditions [182]. Second: P53 has
been reported to mediate proteasomal degradation of HIF-1α [183].
P53 is a redox sensitive protein that can be activated by oxidants
[184,185]. ROS may induce the expression of P53 which in turn pro-
motes the degradation of HIF-1α. Furthermore, HIF-1α, under hypoxic
conditions, may transcriptionally activate its own degradation, likely
mediated through acetylation [185].
As shown by a study, low dose exogenous H2O2 or brief oxygen-
glucose deprivation (OGD) provides neuroprotection against severe
OGD 24 h later. H2O2 induces the expression of HIF-1 α in a dose-de-
pendent manner with a maximal induction at 15 μM. On the other hand,
H2O2 at the dose of 15 μM, causes a time-dependent increase in the
expression of HIF-1α with maximal levels at 32 h after preconditioning
[186].
Emerging evidence demonstrate that in addition to hypoxia (by
inhibiting prolyl hydroxylase-dependent degradation of HIF-1α), H2O2
is also a key factor in the activation of HIF-1 and regulation of its ac-
tivity. This regulation likely depends on the dose of H2O2 and also type
of the cell, as pretreatment with high dose of H2O2 (1 mM) has been
reported to inhibit hypoxia-induced HIF-1α protein accumulation in
human Hep3B cells and Hela cells. It has been shown that non-toxic
levels of H2O2 produced by the enzyme glucose oxidase have no effect
Z. Ghanbari Movahed, et al.
on HIF-1 α stabilization in HT1080 human fibrosarcoma cells
[88,181,187–193].
8. Possible mechanisms of glycolysis deviation to PPP
As previously suggested in this review, cancer cells, when en-
countering ROS, first increase the glycolysis rate and then following the
continuation of oxidative stress their metabolic balance is skewed from
glycolysis pathway to PPP. The possible molecular mechanisms are as
follows:
8.1. The role of PKM2 in glycolysis deviation to PPP
PKM2 is an important molecule involved in the glycolysis deviation
to PPP. Oxidation at Cys358 induced by acute increase in intracellular
ROS, decreases PKM2 activity allowing accumulation of G6P which
subsequently diverts glucose flux to PPP thereby generating sufficient
reducing potential (NADPH) for ROS detoxification [73,193]. There-
fore, PKM2 oxidation at Cys358 confers an advantage to cancer cells
(A549 human lung cancer cells), as it allows cells to tolerate ROS [73]
and promotes cancer cell survival during conditions of oxidative stress
[108].
PKM2 is allosterically regulated allowing switching between high
and low activity states. PKM2 exists in two tetrameric and dimeric
states that are catalytically distinct. Tetrameric PKM2 possesses high
catalytic activity associated with ATP synthesis and catabolic metabo-
lism in cells. Dimeric PKM2, which is the less active state of PKM2, has
low catalytic activity and facilitates the production of glycolytic inter-
mediates to flux into glycolysisbranch pathways including PPP (in-
volved in NADPH and nucleotide synthesis) and glycerol synthesis
pathway [108].
Multiple signals including oncogenes, growth factors and ROS have
been shown to destabilize the tetrameric form via several mechanisms
such as tyrosine phosphorylation, association with tyrosine phospho-
peptides or oxidation [194]. ROS can induce HIF-1 activation and
PKM2 inactivation which subsequently result in increased glycolysis
and PPP rates, respectively.
Studies show that ROS dose and exposure time are two important
factors mediating HIF-1 and PKM2 activation/inactivation.
According to a study, in A549 human lung cancer cells PKM2 is
inactivated in the presence of 250 μM diamide or 1 mM H2O2 for 15 min
[73], while another study showed that HIF-1 is activated in 6–8 days in
vitro (DIV) primary cortical neurons from embryonic day 16 (E16) CD1
mouse brains with 15 μM H2O2 for 10 min [186]. Since activation of
HIF-1 requires less H2O2 dose/exposure time than inactivation of
PKM2, in the face of H2O2, first HIF-1 is activated. Therefore, in re-
sponse to ROS, it is glycolysis which is upregulated first.
With the increase in ROS dose/exposure time, HIF-1 and PKM2 are
inactivated. Oxidation of PKM2 decreases enzyme activity and pro-
motes oxidative PPP flux.
8.2. The role of GAPDH in glycolysis deviation to PPP
Another important molecule involved in the glycolysis deviation to
PPP is GAPDH. ROS inactivates GAPDH by directly targeting cysteine
residues [195]. GAPDH inhibition helps divert glycolytic flux into the
oxidative phase of PPP by allowing metabolites to accumulate upstream
of the inhibition point. HIF-1 activation by ROS increases glycolysis
while ROS-mediated inactivation of GAPDH increases PPP. Studies have
shown that a large amount of ROS is needed to inactivate GAPDH. For
example, in a study, GAPDH was shown to be inactivated with 500 μM
H2O2 for 60 min or 2 mM H2O2 for 30 min in S. cerevisiae exponential-
phase cells [196]. As stated before, HIF-1 is activated with 15 μM H2O2
for 10 min [186]. Therefore, in the face of H2O2, first glycolysis is ac-
tivated and then following the increase in H2O2 dose/exposure time,
HIF-1 and GAPDH are inactivated. GAPDH inactivation leads to
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Biomedicine & Pharmacotherapy 112 (2019) 108690
metabolic switch towards PPP.
8.3. The role of TIGAR in glycolysis deviation to PPP
Another important molecule involved in the glycolysis deviation to
PPP is TIGAR (TP53-induced glycolysis and apoptosis regulator). ROS
and UV stress can trigger TIGAR activation that inhibits PFK1. PFK1
inhibition helps divert glycolytic flux into PPP [125].
In a study, TIGAR was shown to be activated with 720 μmol/L H2O2
for 3 h in A549 human lung cancer cells [197] while as stated before, in
another study HIF-1 was shown to be activated with 15 μM H2O2 for
10 min [186]. Since activation of HIF-1 requires less ROS dose/ex-
posure time than that of TIGAR, HIF-1 is first activated. Therefore, first
glycolysis is upregulated and then following the increase in ROS dose/
exposure time, HIF-1 is inactivated and TIGAR is activated. TIGAR
activation allows glycolytic flux to be diverted into PPP.
8.4. The role of P53 in glycolysis deviation to PPP
P53 is also an important molecule involved in the glycolysis de-
viation to PPP. In response to ROS and UV stress, P53 activates TIGAR.
TIGAR degrades F2,6-BP thereby inhibiting PFK1. This allows glyco-
lytic flux to be diverted into PPP [125].
In a study, it was shown that P53 is activated with 0.6 or 0.8 mM
H2O2 in 293 T cell line [198] Another study showed a slight increase in
the expression of P53 in HeLa cells treated with 125 μM H2O2 in a time
dependent manner (1 h–3 h), but this increase was not statistically
significant [199]. In comparison with P53, HIF-1 activation occurs at
lower H2O2 dose/exposure time [186]. As a consequence, in the face of
H2O2, first glycolysis is upregulated.
9. In response to increased ROS, which one do the cancer cells
increase? PPP or other pathways of NADPH production
Oxidative PPP which is a cytoplasmic branch of glycolysis providing
the cell with both NADPH and ribose for nucleotide synthesis, is
thought to be the main source of NADPH. However, there exist other
potential sources of NADPH such as the reactions involving different
isoforms of isocitrate dehydrogenase (IDH) and malic enzyme (ME).
Though less appreciated (until now), serine and glycine metabolism
exploiting the tetrahydrofolate (THF) cycle, can also be regarded as
notable sources of NADPH [200].
The cytosol possesses several sources of NADPH generation in-
cluding the oxidative PPP, IDH1, malic enzyme 1 (ME1), and one-
carbon metabolism. In the mitochondria, NADPH generation is, in part,
controlled by one-carbon metabolism and IDH2. Cytosolic sources of
NADPH comprise the oxidative PPP, IDH1 and enzymes from one-
carbon metabolism such as methylene tetrahydrofolate dehydrogenase
1 (MTHFD1). Mitochondrial sources of NADPH include IDH2, MTHFD2,
and MTHF2L [201].
There are two major NADPH-generating pathways in the cell. One is
PPP which generates NADPH in the cytoplasm. The first step of PPP,
mediated by G6PD, involves the conversion of the glycolytic inter-
mediate G6P and NADP + to 6-phosphogluconolactone and NADPH.
The other major pathway for NADPH generation is one-carbon meta-
bolism (1CM) which is also known as the folate cycle. Serine is the main
donor of one-carbon units to the folate cycle. The serine synthesis
pathway (SSP) provides a mechanism for the carbons derived from
glucose to be deviated from glycolysis for de novo synthesis of serine
[202].
Serine is required for multiple biosynthetic and signaling pathways
including synthesis of amino acids like cysteine and glycine both of
which are required for glutathione synthesis. In the SSP, the glycolytic
intermediate 3-phosphoglycerate (3-PG) is converted to 3-phospho-
pyruvate (3-PHP) by the action of phosphoglycerate dehydrogenase
(PHGDH). In the next step, phosphoserine aminotransferase (PSAT1),
Z. Ghanbari Movahed, et al.
converts 3-PHP to phosphoserine (P-Ser) and in the final step, phos-
phoserine phosphatase (PSPH) converts P-Ser into serine. Then, serine
and NADP + are utilized in 1CM pathway, either in the cytosol or mi-
tochondria, which produces glycine and NADPH. Mitochondrial 1CM is
catalyzed by the enzymes: serine hydroxymethyltransferase 2 (SHMT2),
MTHFD2, or MTHFD2-like (MTHFD2L), and MTHFD1L. In cytosolic
1CM, the same reactions are mediated by SHMT1 and MTHFD1
(MTHFD1 catalyzes the reactions performed by both MTHFD2/
MTHFD2L and MTHFD1L) [202].
In hypoxia, HIFs activate the transcription of PHGDH, PSAT1, and
PSPH to enhance conversion of glucose to serine; and SHMT2,
MTHFD2, and MTHFD1L to enhance generation of mitochondrial
NADPH (by mitochondrial 1CM), which is required to convert GSSG to
GSH to protect against increased ROS generated by the ETC. Under
hypoxic conditions, HIFs also activate the transcription of solute carrier
family 7 member 11 (SLC7A11) and glutamate-cysteine ligase modifier
subunit (GCLM) to increase glutathione production [203].
In a study, IDH was treated with 1, 2, and 10 mM H2O2 for 12 min.
The results showed that incubation with 1, 2, and 10 mM H2O2 for
12 min dose-dependently decreased IDH activity [204].
HIF-1 activates transcription of PHGDH, PSAT1, and PSPH to in-
crease serine synthesis pathway; and SHMT2, MTHFD2, and MTHFD1L
to increase generation of mitochondrial NADPH. On one hand, HIF-1 is
inactivated at very high H2O2 levels, the conditions that cause the
metabolic balance to be deviated from glycolysis to PPP. On the other
hand, under such high H2O2 concentrations IDH1 activity is decreased
and also due to HIF-1 inactivation transcription of PHGDH, PSAT1,
PSPH, SHMT2, MTHFD2, and MTHFD1L is hindered. Therefore it could
be concluded that under very high ROS levels that cause glycolysis
deviation to PPP, other pathways of NADPH synthesis are down-
regulated.
10. Conclusion
Cancer cells, as compared to normal cells of the body, have higher
levels of ROS. Alteration in glucose metabolism is one of the ways by
which cancer cells decrease the amount of ROS. Cancer cells can de-
crease the amount of ROS by increasing glycolysis and PPP. In this
review, by summarizing the current literature, we showed that cancer
cells, when encountering ROS, first activate the glycolysis pathway and
then following the increase in ROS levels/exposure time, the metabolic
balance is skewed from glycolysis to PPP. What determines this meta-
bolic deviation is probably differential responses of certain proteins to
different ROS exposure conditions (dose and time). For example, HIF-1
activation which upregulates glycolysis, occurs at low doses of ROS
(H2O2) while higher doses of ROS cause HIF-1 inactivation thereby
decreasing glycolysis. Inactivation of PKM2, which causes the deviation
towards PPP, requires higher doses of ROS and longer exposure times
compared to what is required for HIF-1 activation. This confirms that
HIF-1 is active in lower doses and promotes glycolysis, while following
the increase in ROS levels/exposure time HIF-1 and PKM2 proteins are
inactivated consequently decreasing glycolysis and promoting PPP.
Also, increased ROS causes GAPDH inactivation which promotes PPP.
Inactivation of this protein occurs at higher ROS levels and longer ex-
posure times compared to HIF-1 activation. Activation of TIGAR and
P53, which cause the same metabolic deviation, also occur at higher
ROS levels and longer exposure times compared to HIF-1 activation.
Therefore, exposure to higher doses of ROS (with longer exposure
times) downregulates glycolysis and upregulates PPP.
In addition, in this review, we hypothesize that the metabolic de-
viation from glycolysis to PPP might represent alterations in cancer
cells towards CSCs.
CSCs, unlike the differentiated cancer cells that exhibit increased
glycolysis, possess different metabolic characteristics including in-
creased PPP activity. CSCs are responsible for drug resistance, tumor
initiation, metastasis and recurrence, the characteristics that are
11
Biomedicine & Pharmacotherapy 112 (2019) 108690
consistent with the features like increased PPP activity. For example,
increased PPP in cancer cells has been shown to associate with in-
creased drug resistance. Based on the previous findings cancer cells are
inclined to increase PPP rate following exposure to excess amounts of
ROS. Since oxidative stress is a strong inducer of CSCs, the number of
these cells will increase following ROS exposure. Therefore, an increase
in ROS might indicate an increase in the population of CSCs with the
feature of increased PPP. On such a basis, it may seem reasonable to
consider the increase in PPP, its metabolites and related proteins as
markers for the detection of CSC overpopulation. Therefore, con-
sidering this issue could be of significant prognostic value and might
provide insight on more effective treatment of cancer.
ROS, have a double deck sword behaviour in cancer. On the one
hand, ROS can promote protumorigenic signaling which facilitates
proliferation, survival and metastasis of cancer cell. On the other hand,
ROS can promote antitumorigenic signaling and trigger oxidative
stress–induced death in cancer cells. Production of ROS is considered as
a mechanism shared by non-surgical therapeutic approaches for can-
cers, such as radiotherapy and chemotherapy. Therefore, due to the
implication of such therapeutic approaches in triggering cell death, ROS
are also used to kill cancer cells. ROS are increasingly being appreciated
as potent signaling molecules implicated in the regulation of a number
of different biological processes. Cancer cells exhibit higher ROS levels
compared to their normal counterparts partly due to an enhanced
metabolic rate as well as mitochondrial dysfunction. This increased
ROS in cancer cells contributes to molecular and biochemical altera-
tions necessary for tumor initiation, promotion, progression as well as
chemoresistance (protumorigenic effects). However, in cancer cells
further increase in ROS levels to a toxic level, through augmenting
chemosensitization and induction of various cell-death pathways, pro-
vides an avenue for cancer treatment (antitumorigenic effects).
Therefore, biological effects of ROS in cancer are complex, paradoxical
and context-dependent. The dual aspects of ROS in cancer biology
provides us with a rational to develop two different contradicting ap-
proaches for cancer treatment. The first strategy involves lowering in-
tracellular ROS levels in cancer cells either by inhibiting ROS genera-
tion or by augmenting ROS scavenging activities using different
antioxidants. For instance, metformin, as an anti-diabetic agent, has
been shown to inhibit mitochondrial respiratory chain complex I within
cancer cells thereby reducing tumorigenesis. Also included in this
strategy, we have the use of antioxidants, either alone or in combina-
tion with other antitumor agents, as a promosing alternative to decrease
intracellular ROS-mediated tumorigenesis [39]. Based on the pivotal
role ROS play as a key constituent in tumor biology [38] as well as
considering our hypothesis on the contribution of higher ROS levels to
CSC development, lowering ROS using antioxidants seems to be a
promising approach for cancer targeting. However, it is a challenging
approach to adopt. Even though antioxidant therapy combats cancer
through impeding ROS-induced carcinogenesis, it can also have pro-
tumor effects by inhibiting cellular senescence or apoptosis induced by
higher ROS levels as a consequence of high genomic instability of
cancer cells [205].
The second ROS-mediated strategy for cancer treatment is to in-
crease ROS levels in the cancer cell to a toxic level able to activate ROS-
induced cell death pathways. This strategy can be achieved through the
use of agents that either augment ROS generation or inhibit antioxidant
systems, or a combination of both. Regarding the fact that in cancer cell
there is an already increased level of ROS compared to its normal
counterpart, therapeutic interventions further increasing the generation
of ROS can specifically induce cell death in cancer cell. For example, it
has been shown that beta-phenylethyl isothiocyanate and 2-methox-
yoestradiol, by causing further increase in ROS, selectively induce
death in human leukemia cells over normal lymphocytes. However, in
this respect eradicating cancer cells does not seem that easy. Cancer
cells also possess higher antioxidant capabilities to prevent excessive
accumulation of ROS [39].
Z. Ghanbari Movahed, et al.
Based on the current review, we propose that treatments based on
oxidative stress can not be considered as appropriate cancer treatment
strategies, as these treatments could induce adaptive mechanisms in
cancer cells, mechanisms that cause the deviation from glycolysis to
PPP which could be translated as a step from cancer cell to CSC. This
point of view further complicates ROS-mediated therapeutic ap-
proaches in tumor context included in the second strategy and adds to
the existing complexity and controversial aspects of ROS exploitation
for cancer treatment. Considering this hypothesis, if we want to apply
the second strategy (activation of ROS-induced cell death pathways) to
target cancer cells, ROS-inducing agents are suggested to be used in
combination with antioxidant inhibitors in order to cope with CSCs
(with high antioxidant capabilities) that are likely to develop following
our therapeutic intervention. Also combined with ROS-promoting
agents, we interestingly suggest the use of oxidative PPP inhibitors; for
example molecules that inhibit G6PD or 6PGD. It is also suggested to
highly focus on therapies specifically targeting important molecules
mediating glycolysis deviation to PPP in combination with ROS-pro-
moting agents. However, achieving a well-tailored ROS-mediated
therapeutic approach for cancer treatment involves a tremendous and
detailed knowledge of dual aspects of ROS in tumor biology and calls
for further investigations in this amazing field of cancer research.
Conflict of interest
The authors declare that they have no conflict of interest.
Acknowledgments
This research was supported by Medical Biology Research Center,
Kermanshah University of Medical Sciences, Kermanshah, Iran (96599).
We would like to specially thank to Prof. Reza Khodarahmi, Dr Bizhan
Solimani, Dr Hamid Rza Mohammadimotlagh and Mozhgan Jahani for
their valuable discussions and help with manuscript preparation.
References
[1] E. Panieri, M. Santoro, ROS homeostasis and metabolism: a dangerous liason in
cancer cells, Cell Death Dis. 7 (2016) e2253.
[2] G.Y. Liou, P. Storz, Reactive oxygen species in cancer, Free Radic. Res. 44 (2010)
479–496.
[3] I.M. Ahmad, N. Aykin-Burns, J.E. Sim, S.A. Walsh, R. Higashikubo, G.R. Buettner,
S. Venkataraman, M.A. Mackey, S.W. Flanagan, L.W. Oberley, D.R. Spitz,
Mitochondrial and H2O2 mediate glucose deprivation-induced stress in human
cancer cells, J. Biol. Chem. 280 (2005) 4254–4263.
[4] N. Aykin-Burns, I.M. Ahmad, Y. Zhu, L.W. Oberley, D.R. Spitz, Increased levels of
superoxide and H2O2 mediate the differential susceptibility of cancer cells versus
normal cells to glucose deprivation, Biochem. J. 418 (2009) 29–37.
[5] D. Zhou, L. Shao, D.R. Spitz, Reactive oxygen species in normal and tumor stem
cells, Adv. Cancer Res. 122 (2014) 1–67.
[6] D.R. Spitz, J.E. Sim, L.A. Ridnour, S.S. Galoforo, Y.J. Lee, Glucose deprivation-
induced oxidative stress in human tumor cells: a fundamental defect in metabo-
lism? Ann. N. Y. Acad. Sci. 899 (2000) 349–362.
[7] O. Vafa, M. Wade, S. Kern, M. Beeche, T.K. Pandita, G.M. Hampton, G.M. Wahl, c-
Myc can induce DNA damage, increase reactive oxygen species, and mitigate p53
function: a mechanism for oncogene-induced genetic instability, Mol. Cell. 9
(2002) 1031–1044.
[8] V. Nogueira, Y. Park, C.C. Chen, P.Z. Xu, M.-L. Chen, I. Tonic, T. Unterman,
N. Hay, Akt determines replicative senescence and oxidative or oncogenic pre-
mature senescence and sensitizes cells to oxidative apoptosis, Cancer Cell 14
(2008) 458–470.
[9] A.A. Sablina, A.V. Budanov, G.V. Ilyinskaya, L.S. Agapova, J.E. Kravchenko,
P.M. Chumakov, The antioxidant function of the p53 tumor suppressor, Nat. Med.
11 (2005) 1306.
[10] K. Bensaad, E.C. Cheung, K.H. Vousden, Modulation of intracellular ROS levels by
TIGAR controls autophagy, EMBO J. 28 (2009) 3015–3026.
[11] W. Hu, C. Zhang, R. Wu, Y. Sun, A. Levine, Z. Feng, Glutaminase 2, a novel p53
target gene regulating energy metabolism and antioxidant function, Proc. Natl.
Acad. Sci. 107 (2010) 7455–7460.
[12] S. Reuter, S.C. Gupta, M.M. Chaturvedi, B.B. Aggarwal, Oxidative stress, in-
flammation, and cancer: how are they linked? Free Radic. Biol. Med. 49 (2010)
1603–1616.
[13] B. Halliwell, Oxidative stress and cancer: have we moved forward? Biochem. J.
401 (2007) 1–11.
12
Biomedicine & Pharmacotherapy 112 (2019) 108690
[14] Z.T. Schafer, A.R. Grassian, L. Song, Z. Jiang, Z. Gerhart-Hines, H.Y. Irie, S. Gao,
P. Puigserver, J.S. Brugge, Antioxidant and oncogene rescue of metabolic defects
caused by loss of matrix attachment, Nature 461 (2009) 109.
[15] K. Ishikawa, K. Takenaga, M. Akimoto, N. Koshikawa, A. Yamaguchi, H. Imanishi,
K. Nakada, Y. Honma, J. Hayashi, ROS-generating mitochondrial DNA mutations
can regulate tumor cell metastasis, Science 320 (2008) 661–664.
[16] F. Weinberg, R. Hamanaka, W.W. Wheaton, S. Weinberg, J. Joseph, M. Lopez,
B. Kalyanaraman, G.M. Mutlu, G.R. Budinger, N.S. Chandel, Mitochondrial me-
tabolism and ROS generation are essential for Kras-mediated tumorigenicity, Proc.
Natl. Acad. Sci. U. S. A. 107 (2010) 8788–8793.
[17] D. Trachootham, J. Alexandre, P. Huang, Targeting cancer cells by ROS-mediated
mechanisms: a radical therapeutic approach? Nat. Rev. Drug Discov. 8 (2009) 579.
[18] A. Glasauer, N.S. Chandel, Targeting antioxidants for cancer therapy, Biochem.
Pharmacol. 92 (2014) 90–101.
[19] G.M. DeNicola, F.A. Karreth, T.J. Humpton, A. Gopinathan, C. Wei, K. Frese,
D. Mangal, K.H. Yu, C.J. Yeo, E.S. Calhoun, F. Scrimieri, J.M. Winter, R.H. Hruban,
C. Iacobuzio-Donahue, S.E. Kern, I.A. Blair, D.A. Tuveson, Oncogene-induced Nrf2
transcription promotes ROS detoxification and tumorigenesis, Nature 475 (2011)
106.
[20] S. Kamarajugadda, L. Stemboroski, Q. Cai, N.E. Simpson, S. Nayak, M. Tan, J. Lu,
Glucose oxidation modulates anoikis and tumor metastasis, Mol. Cell. Biol. 32
(2012) 1893–1907.
[21] P. Jiang, W. Du, M. Wu, Regulation of the pentose phosphate pathway in cancer,
Protein Cell 5 (2014) 592–602.
[22] J. Xie, H. Wu, C. Dai, Q. Pan, Z. Ding, D. Hu, B. Ji, Y. Luo, X. Hu, Beyond Warburg
effect–dual metabolic nature of cancer cells, Sci. Rep. 4 (2014) 4927.
[23] C.E. Meacham, S.J. Morrison, Tumour heterogeneity and cancer cell plasticity,
Nature 501 (2013) 328.
[24] Q. Pan, Q. Li, S. Liu, N. Ning, X. Zhang, Y. Xu, A.E. Chang, M.S. Wicha, Concise
review: targeting cancer stem cells using immunologic approaches, Stem Cells 33
(2015) 2085–2092.
[25] B.G. Debeb, L. Lacerda, R. Larson, A.R. Wolfe, S. Krishnamurthy, J.M. Reuben,
N.T. Ueno, M. Gilcrease, W.A. Woodward, Histone deacetylase inhibitor-induced
cancer stem cells exhibit high pentose phosphate pathway metabolism, Oncotarget
7 (2016) 28329.
[26] K.C. Patra, N. Hay, The phosphate pathway and cancer, Trends Biochem. Sci. 39
(2014) 347–354.
[27] T. Reya, S.J. Morrison, M.F. Clarke, I.L. Weissman, Stem cells, cancer, and cancer
stem cells, Nature (2001) 105.
[28] Y.J. Kim, E.L. Siegler, N. Siriwon, P. Wang, Therapeutic strategies for targeting
cancer stem cells, J. Cancer Metastasis Treat. 2 (2016) 234.
[29] D.L. Dragu, L.G. Necula, C. Bleotu, C.C. Diaconu, M. Chivu-Economescu, Therapies
targeting cancer stem cells: current trends and future challenges, World J. Stem
Cells 7 (2015) 1185.
[30] H.J. Forman, O. Augusto, R. Brigelius-Flohe, P.A. Dennery, B. Kalyanaraman,
H. Ischiropoulos, G.E. Mann, R. Radi, L.J. Roberts, J. Vina, K.J. Davies, Even free
radicals should follow some rules: a guide to free radical research terminology and
methodology, Free Radic. Biol. Med. 78 (2015) 233–235.
[31] H.J. Forman, M. Maiorino, F. Ursini, Signaling functions of reactive oxygen spe-
cies, Biochemistry 49 (2010) 835–842.
[32] C.C. Winterbourn, M.B. Hampton, Thiol chemistry and specificity in redox sig-
naling, Free Radic. Biol. Med. 45 (2008) 549–561.
[33] H. Sies, Hydrogen peroxide as a central redox signaling molecule in physiological
oxidative stress: oxidative eustress, Redox Biol. 11 (2017) 613–619.
[34] E.I. Azzam, J.P. Jay-Gerin, D. Pain, Ionizing radiation-induced metabolic oxidative
stress and prolonged cell injury, Cancer Lett. 327 (2012) 48–60.
[35] W. Droge, Free radicals in the physiological control of cell function, Physiol. Rev.
82 (2002) 47–95.
[36] W. Pendergrass, G. Zitnik, R. Tsai, N. Wolf, X-ray induced cataract is preceded by
LEC loss, and coincident with accumulation of cortical DNA, and ROS; similarities
with age-related cataracts, Mol. Vis. 16 (2010) 1496.
[37] P. Riley, Free radicals in biology: oxidative stress and the effects of ionizing ra-
diation, Int. J. Radiat. Biol. 65 (1994) 27–33.
[38] S. Kumari, A.K. Badana, M.M. G, S. G, R. Malla, Reactive Oxygen Species: A Key
Constituent in Cancer Survival, Biomark. Insights 13 (2018) 1177271918755391..
[39] S. Galadari, A. Rahman, S. Pallichankandy, F. Thayyullathil, Reactive oxygen
species and cancer paradox: to promote or to suppress? Free Radic. Biol. Med. 104
(2017) 144–164.
[40] I. Ganan-Gomez, Y. Wei, H. Yang, M.C. Boyano-Adanez, G. Garcia-Manero,
Oncogenic functions of the transcription factor Nrf2, Free Radic. Biol. Med. 65
(2013) 750–764.
[41] T.P. Szatrowski, C.F. Nathan, Production of large amounts of hydrogen peroxide
by human tumor cells, Cancer Res. 51 (1991) 794–798.
[42] A. Costa, A. Scholer-Dahirel, F. Mechta-Grigoriou, The role of reactive oxygen
species and metabolism on cancer cells and their microenvironment, Semin.
Cancer Biol. 25 (2014) 23–32.
[43] W. Li, L. Cao, L. Han, Q. Xu, Q. Ma, Superoxide dismutase promotes the epithelial-
mesenchymal transition of pancreatic cancer cells via activation of the H2O2/
ERK/NF-κB axis, Int. J. Oncol. 46 (2015) 2613–2620.
[44] M.C. Kim, F.J. Cui, Y. Kim, Hydrogen peroxide promotes epithelial to mesench-
ymal transition and stemness in human malignant mesothelioma cells, Asian Pac.
J. Cancer Prev. 14 (2013) 3625–3630.
[45] V. Ribas, C. Garcia-Ruiz, J.C. Fernandez-Checa, Mitochondria, cholesterol and
cancer cell metabolism, Clin. Transl. Med. 5 (2016) 22.
[46] A. Federico, F. Morgillo, C. Tuccillo, F. Ciardiello, C. Loguercio, Chronic in-
flammation and oxidative stress in human carcinogenesis, Int. J. Cancer 121
Z. Ghanbari Movahed, et al.
(2007) 2381–23896.
[47] M. Fransen, M. Nordgren, B. Wang, O. Apanasets, Role of peroxisomes in ROS/
RNS-metabolism: implications for human disease, Biochim. Biophys. Acta. Mol.
Basis Dis. 1822 (2012) 1363–1373.
[48] V.D. Antonenkov, S. Grunau, S. Ohlmeier, J.K. Hiltunen, Peroxisomes are oxida-
tive organelles, Antioxid. Redox Signal. 13 (2010) 525–537.
[49] M. Valko, C. Rhodes, J. Moncol, M. Izakovic, M. Mazur, Free radicals, metals and
antioxidants in oxidative stress-induced cancer, Chem.-Biol. Interact. 160 (2006)
1–40.
[50] M. Inoue, E.F. Sato, M. Nishikawa, A.M. Park, Y. Kira, I. Imada, Utsumi K,
Mitochondrial generation of reactive oxygen species and its role in aerobic life,
Curr. Med. Chem. 10 (2003) 2495–2505.
[51] D.E. Handy, J. Loscalzo, Redox regulation of mitochondrial function, Antioxid.
Redox Signal. 16 (2012) 1323–1367.
[52] M.M. Briehl, Oxygen in human health from life to death–an approach to teaching
redox biology and signaling to graduate and medical students, Redox Boil 5 (2015)
124–139.
[53] M.P. Murphy, How mitochondria produce reactive oxygen species, Biochem. J.
417 (2009) 1–13.
[54] D.C. Liemburg-Apers, P.H. Willems, W.J. Koopman, S. Grefte, Interactions be-
tween mitochondrial reactive oxygen species and cellular glucose metabolism,
Arch. Toxicol. 89 (2015) 1209–1226.
[55] F. Cosentino, P. Francia, G.G. Camici, P.G. Pelicci, M. Volpe, T.F. Luscher, Final
common molecular pathways of aging and cardiovascular disease: role of the
p66Shc protein, Arterioscler. Thromb. Vasc. Biol. 28 (2008) 622–628.
[56] K.M. Holmstrom, Finkel T, Cellular mechanisms and physiological consequences
of redox-dependent signalling, Nat. Rev. Mol. Cell Biol. 15 (2014) 411.
[57] N.Y. Spencer, J.F. Engelhardt, The basic biology of redoxosomes in cytokine-
mediated signal transduction and implications for disease-specific therapies,
Biochemistry 53 (2014) 1551–1564.
[58] E.E. Kelley, N.K. Khoo, N.J. Hundley, U.Z. Malik, B.A. Freeman, M.M. Tarpey,
Hydrogen peroxide is the major oxidant product of xanthine oxidase, Free Radical.
Bio. Med. 48 (2010) 493–498.
[59] I. Fridovich, Quantitative aspects of the production of superoxide anion radical by
milk xanthine oxidase, J. Biol. Chem. 245 (1970) 4053–4057.
[60] M. Tafani, L. Sansone, F. Limana, T. Arcangeli, E. De Santis, M. Polese, F. Massimo,
A.R. Matteo, The interplay of reactive oxygen species, hypoxia, inflammation, and
sirtuins in cancer initiation and progression, Oxid. Med. Cell. Longev. (2016).
[61] M. Allard, B. Schonekess, S. Henning, D. English, G.D. Lopaschuk, Contribution of
oxidative metabolism and glycolysis to ATP production in hypertrophied hearts,
Am. J. Physiol. Heart Circ. Physiol. 267 (1994) 742–750.
[62] M.G. Vander Heiden, L.C. Cantley, C.B. Thompson, Understanding the Warburg
effect: the metabolic requirements of cell proliferation, Science 324 (2009)
1029–1033.
[63] D. Nelson, Lehninger Principles of Biochemistry, 5nd edn, Macmillan, 2008.
[64] M. Jahani, F. Noroznezhad, K. Mansouri, Arginine: challenges and opportunities of
this two-faced molecule in cancer therapy, Biomed. Pharmacother. 102 (2018)
594–601.
[65] O. Warburg, On the origin of cancer cells, Science 123 (1956) 309–314.
[66] S.W. House, O. Warburg, D. Burk, A.L. Schade, On respiratory impairment in
cancer cells, Science 124 (1956) 267–272.
[67] Warburg O, Minami S, Versuche an uberlebendem carcinom-gewebe, J. Mol. Med.
2 (1923) 776–777.
[68] O. Warburg, Uber den Stoffwechsel der Carcinomzelle, Klin. Wochenschr. 4 (1925)
534–536.
[69] E. Racker, Bioenergetics and the problem of tumor growth: an understanding of
the mechanism of the generation and control of biological energy may shed light
on the problem of tumor growth, Am. Sci. 60 (1972) 56–63.
[70] J.C. Kotz, P.M. Treichel, J. Townsend, Chemistry and Chemical Reactivity, 8 ed.,
Cengage Learning, 2012.
[71] X. Tong, F. Zhao, C.B. Thompson, The molecular determinants of de novo nu-
cleotide biosynthesis in cancer cells, Curr. Opin. Genet. Dev. 19 (2009) 32–37.
[72] J.W. Locasale, A.R. Grassian, T. Melman, C.A. Lyssiotis, K.R. Mattaini, A.J. Bass,
G. Heffron, C.M. Metallo, T. Muranen, H. Sharfi, A.T. Sasaki, D. Anastasiou,
E. Mullarky, N.I. Vokes, M. Sasaki, R. Beroukhim, G. Stephanopoulos, A.H. Ligon,
M. Meyerson, A.L. Richardson, L. Chin, G. Wagner, J.M. Asara, J.S. Brugge,
L.C. Cantley, M.G. Vander Heiden, Phosphoglycerate dehydrogenase diverts gly-
colytic flux and contributes to oncogenesis, Nat. Genet. 43 (2011) 869.
[73] D. Anastasiou, G. Poulogiannis, J.M. Asara, M.B. Boxer, Jk. Jiang, M. Shen,
G. Bellinger, A.T. Sasaki, J.W. Locasale, D.S. Auld, C.J. Thomas, M.G. Vander
Heiden, L.C. Cantley, Inhibition of pyruvate kinase M2 by reactive oxygen species
contributes to cellular antioxidant responses, Science 334 (2011) 1278–1283.
[74] R.B. Hamanaka, N.S. Chandel, Warburg effect and redox balance, Science 334
(2011) 1219–1220.
[75] M. Lee, J.H. Yoon, Metabolic interplay between glycolysis and mitochondrial
oxidation: the reverse Warburg effect and its therapeutic implication, World J.
Biol. Chem. 6 (2015) 148.
[76] L.L. Grochowski, H. Xu, R.H. White, Ribose-5-phosphate biosynthesis in
Methanocaldococcus jannaschii occurs in the absence of a pentose-phosphate
pathway, J. Bacteriol. 187 (2005) 7382–7389.
[77] T. Nunoura, Y. Takaki, J. Kakuta, S. Nishi, J. Sugahara, H. Kazama, G.J. Chee,
M. Hattori, A. Kanai, H. Atomi, K. Takai, H. Takami, Insights into the evolution of
Archaea and eukaryotic protein modifier systems revealed by the genome of a
novel archaeal group, Nucleic Acids Res. 39 (2010) 3204–3223.
[78] C. Brasen, D. Esser, B. Rauch, B. Siebers, Carbohydrate metabolism in Archaea:
current insights into unusual enzymes and pathways and their regulation,
13
Biomedicine & Pharmacotherapy 112 (2019) 108690
Microbiol. Mol. Biol. Rev. 78 (2014) 89–175.
[79] A. Stincone, A. Prigione, T. Cramer, M. Wamelink, K. Campbell, E. Cheung,
V. Olin-Sandoval, N.M. Gruning, A. Kruger, M. Tauqeer Alam, M.A. Keller,
M. Breitenbach, K.M. Brindle, J.D. Rabinowitz, M. Ralser, The return of metabo-
lism: biochemistry and physiology of the pentose phosphate pathway, Biol Rev. 90
(2015) 927–963.
[80] P.J. Fernandez-Marcos, S. Nobrega-Pereira, NADPH: new oxygen for the ROS
theory of aging, Oncotarget 7 (2016) 50814.
[81] R.B. Hamanaka, N.S. Chandel, Targeting glucose metabolism for cancer therapy, J.
Exp. Med. 209 (2012) 211–215.
[82] C.M. Labak, P.Y. Wang, R. Arora, M.R. Guda, S. Asuthkar, A.J. Tsung, K.V. Kiran,
Glucose transport: meeting the metabolic demands of cancer, and applications in
glioblastoma treatment, Am. J. Cancer Res. 6 (2016) 1599.
[83] T.B. Zhang, Y. Zhao, Z.X. Tong, Y.F. Guan, Inhibition of glucose-transporter 1
(GLUT-1) expression reversed Warburg effect in gastric cancer cell MKN45, Int. J.
Clin. Exp. Med. 8 (2015) 2423.
[84] S. Andrisse, G.D. Patel, J.E. Chen, A.M. Webber, L.D. Spears, R.M. Koehler,
Ching Robinson-Hill, R.M, J.K, I. Jeong, J.S, ATM and GLUT1-S490 phosphor-
ylation regulate GLUT1 mediated transport in skeletal muscle, PLoS One 8 (2013)
e66027.
[85] A. Alexander, C.L. Walker, Differential localization of ATM is correlated with ac-
tivation of distinct downstream signaling pathways, Cell Cycle 9 (2010)
3709–3710.
[86] G.L. Semenza, Hypoxia-inducible factor 1: master regulator of O 2 homeostasis,
Curr. Opin. Genet. Dev. 8 (1998) 588–594.
[87] G.L. Wang, G.L. Semenza, Purification and characterization of hypoxia-inducible
factor 1, J. Biol. Chem. 270 (1995) 1230–1237.
[88] L.E. Huang, Z. Arany, D.M. Livingston, H.F. Bunn, Activation of hypoxia-inducible
transcription factor depends primarily upon redox-sensitive stabilization of its α
subunit, J. Biol. Chem. 271 (1996) 32253–32259.
[89] R.S. Bel Aiba, E.Y. Dimova, A. Gorlach, T. Kietzmann, The role of hypoxia in-
ducible factor-1 in cell metabolism–a possible target in cancer therapy, Expert
Opin. Ther. Targets 10 (2006) 583–599.
[90] M. Jahani, K. Mansouri, Role of hypoxia and hypoxia inducible factor in physio-
logical and pathological conditions, J. Kermanshah Univ. Med Sci. 21 (2017)
126–132.
[91] G.N. Masoud, W. Li, HIF-1α pathway: role, regulation and intervention for cancer
therapy, Acta Pharm. Sin. B 5 (2015) 378–389.
[92] K. Mansouri, A. Mostafie, D. Rezazadeh, M. Shahlaei, M.H. Modarressi, New
function of TSGA10 gene in angiogenesis and tumor metastasis: a response to a
challengeable paradox, Hum. Mol. Genet. 25 (2015) 233–244.
[93] P.E. Porporato, R.K. Dadhich, S. Dhup, T. Copetti, P. Sonveaux, Anticancer targets
in the glycolytic metabolism of tumors: a comprehensive review, Front.
Pharmacol. 2 (2011) 49.
[94] S.P. Mathupala, A. Rempel, P.L. Pedersen, Glucose catabolism in cancer cells
identification and characterization of a marked activation response of the type II
hexokinase gene to hypoxic conditions, J. Biol. Chem. 276 (2001) 43407–43412.
[95] D.A. Okar, A.J. Lange, Fructose‐2, 6‐bisphosphate and control of carbohydrate
metabolism in eukaryotes, Biofactors 10 (1999) 1–14.
[96] D.A. Okar, A.J. Lange, A. Manzano, A. Navarro-Sabate, Ls. Riera, R. Bartrons, PFK-
2/FBPase-2: maker and breaker of the essential biofactor fructose-2, 6-bispho-
sphate, Trends Biochem. Sci. 26 (2001) 30–35.
[97] A. Minchenko, I. Leshchinsky, I. Opentanova, N. Sang, V. Srinivas, V. Armstead,
J. Caro, Hypoxia-inducible Factor-1-mediated Expression of the 6-Phosphofructo-
2-kinase/fructose-2, 6-bisphosphatase-3 (PFKFB3) Gene its possible role in the
warburg effect, J. Biol. Chem. 277 (2002) 6183–6187.
[98] O. Minchenko, I. Opentanova, J. Caro, Hypoxic regulation of the 6‐phospho-
fructo‐2‐kinase/fructose‐2, 6‐bisphosphatase gene family (PFKFB‐1–4) expression
in vivo, FEBS Lett. 554 (2003) 264–270.
[99] Jw. Kim, I. Tchernyshyov, G.L. Semenza, C.V. Dang, HIF-1-mediated expression of
pyruvate dehydrogenase kinase: a metabolic switch required for cellular adapta-
tion to hypoxia, Cell Metab. 3 (2006) 177–185.
[100] I. Papandreou, R.A. Cairns, L. Fontana, A.L. Lim, N.C. Denko, HIF-1 mediates
adaptation to hypoxia by actively downregulating mitochondrial oxygen con-
sumption, Cell Metab. 3 (2006) 187–197.
[101] S. Wigfield, S. Winter, A. Giatromanolaki, J. Taylor, M. Koukourakis, A. Harris,
PDK-1 regulates lactate production in hypoxia and is associated with poor prog-
nosis in head and neck squamous cancer, Br. J. Cancer 98 (2008) 1975.
[102] C.L. Markert, J.B. Shaklee, G.S. Whitt, Evolution of a gene, Science 189 (1975)
102–114.
[103] M. Koukourakis, A. Giatromanolaki, E. Sivridis, G. Bougioukas, V. Didilis,
K. Gatter, A.L. Harris, Lactate dehydrogenase-5 (LDH-5) overexpression in non-
small-cell lung cancer tissues is linked to tumour hypoxia, angiogenic factor
production and poor prognosis, Br. J. Cancer 89 (2003) 877.
[104] M.I. Koukourakis, A. Giatromanolaki, E. Sivridis, K.C. Gatter, T. Trarbach,
G. Folprecht, M.M. Shi, D. Lebwohl, T. Jalava, D. Laurent, G. Meinhardt,
A.L. Harris, Prognostic and predictive role of lactate dehydrogenase 5 expression
in colorectal cancer patients treated with PTK787/ZK 222584 (vatalanib) anti-
angiogenic therapy, Clin. Cancer Res. 17 (2011) 4892–4900.
[105] M.I. Koukourakis, A. Giatromanolaki, S. Winter, R. Leek, E. Sivridis, A.L. Harris,
Lactate dehydrogenase 5 expression in squamous cell head and neck cancer relates
to prognosis following radical or postoperative radiotherapy, Oncology 77 (2009)
285–292.
[106] A. Leiblich, S. Cross, J. Catto, J. Phillips, H. Leung, F. Hamdy, A.L. Harris, Lactate
dehydrogenase-B is silenced by promoter hypermethylation in human prostate
cancer, Oncogene 25 (2006) 2953.
Z. Ghanbari Movahed, et al.
[107] M. Thangaraju, K.N. Carswell, P.D. Prasad, V. Ganapathy, Colon cancer cells
maintain low levels of pyruvate to avoid cell death caused by inhibition of
HDAC1/HDAC3, Biochem. J. 417 (2009) 379–389.
[108] G. Dong, Q. Mao, W. Xia, Y. Xu, J. Wang, L. Xu, F. Jiang, PKM2 and cancer: the
function of PKM2 beyond glycolysis, Oncol. Lett. 11 (2016) 1980–1986.
[109] G.L. Semenza, HIF-1 mediates metabolic responses to intratumoral hypoxia and
oncogenic mutations, J. Clin. Invest. 123 (2013) 3664–3671.
[110] G.N. Masoud, W. Li, HIF-1α pathway: role, regulation and intervention for cancer
therapy, Acta Pharm. Sin. B 5 (2015) 378–389.
[111] S.N. Jung, W.K. Yang, J. Kim, H.S. Kim, E.J. Kim, H. Yun, H. Park, S.S. Kim,
W. Choe, I. Kang, J. Ha, Reactive oxygen species stabilize hypoxia-inducible
factor-1 alpha protein and stimulate transcriptional activity via AMP-activated
protein kinase in DU145 human prostate cancer cells, Carcinogenesis 29 (2008)
713–721.
[112] Dy. Shi, Fz. Xie, C. Zhai, J.S. Stern, Y. Liu, Sl. Liu, The role of cellular oxidative
stress in regulating glycolysis energy metabolism in hepatoma cells, Mol. Cancer 8
(2009) 32.
[113] T. Hitosugi, S. Kang, M.G. Vander Heiden, T.W. Chung, S. Elf, K. Lythgoe, S. Dong,
S. Lonial, X. Wang, G.Z. Chen, J. Xie, T.L. Gu, R.D. Polakiewicz, J.L. Roesel,
T.J. Boggon, F.R. Khuri, D.G. Gilliland, L.C. Cantley, J. Kaufman, J. Chen, Tyrosine
phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth,
Sci. Signal. 2 (2009) ra73.
[114] H.R. Christofk, M.G. Vander Heiden, M.H. Harris, A. Ramanathan, R.E. Gerszten,
R. Wei, M.D. Fleming, S.L. Schreiber, L.C. Cantley, The M2 splice isoform of
pyruvate kinase is important for cancer metabolism and tumour growth, Nature
452 (2008) 230.
[115] M.G. Vander Heiden, J.W. Locasale, K.D. Swanson, H. Sharfi, G.J. Heffron,
D. Amador-Noguez, H.R. Christofk, G. Wagner, J.D. Rabinowitz, J.M. Asara,
L.C. Cantley, Evidence for an alternative glycolytic pathway in rapidly pro-
liferating cells, Science 329 (2010) 1492–1499.
[116] N.M. Gruning, M. Rinnerthaler, K. Bluemlein, M. Mulleder, M.M. Wamelink,
H. Lehrach, C. Jakobs, M. Breitenbach, M. Ralser, Pyruvate kinase triggers a me-
tabolic feedback loop that controls redox metabolism in respiring cells, Cell Metab.
14 (2011) 415–427.
[117] R.A. Cairns, I.S. Harris, T.W. Mak, Regulation of cancer cell metabolism, Nat. Rev.
Cancer 11 (2011) 85.
[118] A. Kruger, N.M. Gruning, M.M. Wamelink, M. Kerick, A. Kirpy, D. Parkhomchuk,
K. Bluemlein, M.R. Schweiger, A. Soldatov, H. Lehrach, C. Jakobs, M. Ralser, The
pentose phosphate pathway is a metabolic redox sensor and regulates transcrip-
tion during the antioxidant response, Antioxid. Redox Signal. 15 (2011) 311–324.
[119] J.K. Brunelle, E.L. Bell, N.M. Quesada, K. Vercauteren, V. Tiranti, M. Zeviani,
R.C. Scarpulla, N.S. Chandel, Oxygen sensing requires mitochondrial ROS but not
oxidative phosphorylation, Cell Metab. 1 (2005) 409–414.
[120] M. Ralser, M.M. Wamelink, A. Kowald, B. Gerisch, G. Heeren, E.A. Struys, E. Klipp,
C. Jakobs, M. Breitenbach, H. Lehrach, S. Krobitsch, Dynamic rerouting of the
carbohydrate flux is key to counteracting oxidative stress, J. Biol. 6 (2007) 10.
[121] D. Shenton, C.M. Grant, Protein S-thiolation targets glycolysis and protein
synthesis in response to oxidative stress in the yeast Saccharomyces cerevisiae,
Biochem. J. 374 (2003) 513–519.
[122] C. Godon, G. Lagniel, J. Lee, J.M. Buhler, S. Kieffer, M. Perrot, H. Boucherie,
M.B. Toledano, J. Labarre, The H2O2 stimulon in Saccharomyces cerevisiae, J.
Biol. Chem. 273 (1998) 22480–22489.
[123] C.G. Cochrane, Cellular injury by oxidants, Am. J. Med. 91 (1991) 23–30.
[124] P. Hyslop, D. Hinshaw, Jr.W. Halsey, I. Schraufstatter, R. Sauerheber, R. Spragg,
J.H. Jackson, C.G. Cochrane, The glycolytic and mitochondrial pathways of ADP
phosphorylation are major intracellular targets inactivated by hydrogen peroxide,
J. Biol. Chem. 263 (1988) 1665–1675.
[125] E. Mullarky, L.C. Cantley, Diverting glycolysis to combat oxidative stress,
InnoMed. 2015 (2015) 3–23.
[126] M.A. Kowalik, A. Columbano, A. Perra, Emerging role of the pentose phosphate
pathway in hepatocellular carcinoma, Front. Oncol. 7 (2017) 87.
[127] N. Verma, M. Pink, S. Boland, A.W. Rettenmeier, S. Schmitz-Spanke, Benzo [a]
pyrene-induced metabolic shift from glycolysis to pentose phosphate pathway in
the human bladder cancer cell line RT4, Sci. Rep. 7 (2017) 9773.
[128] E. Vlashi, C. Lagadec, L. Vergnes, T. Matsutani, K. Masui, M. Poulou, R. Popescu,
L. Della Donna, P. Evers, C. Dekmezian, K. Reue, H. Christofk, P.S. Mischel,
F. Pajonk, Metabolic state of glioma stem cells and nontumorigenic cells, Proc.
Natl. Acad. Sci. U. S. A. 108 (2011) 16062–16067.
[129] E.D. Lagadinou, A. Sach, K. Callahan, R.M. Rossi, S.J. Neering, M. Minhajuddin,
J.M. Ashton, S. Pei, V. Grose, K.M. O’Dwyer, J.L. Liesveld, P.S. Brookes,
M.W. Becker, C.T. Jordan, BCL-2 inhibition targets oxidative phosphorylation and
selectively eradicates quiescent human leukemia stem cells, Cell Stem Cell 12
(2013) 329–341.
[130] E. Vlashi, C. Lagadec, L. Vergnes, K. Reue, P. Frohnen, M. Chan, Y. Alhiyari,
M.B. Dratver, F. Pajonk, Metabolic differences in breast cancer stem cells and
differentiated progeny, Breast Cancer Res. Treat. 146 (2014) 525–534.
[131] D. Ciavardelli, C. Rossi, D. Barcaroli, S. Volpe, A. Consalvo, M. Zucchelli, A. De
Cola, E. Scavo, R. Carollo, D. DAgostino, F. ForlI, S. DAguanno, M. Todaro,
G. Stassi, C. Di Ilio, V. De Laurenzi, A. Urbani, Breast cancer stem cells rely on
fermentative glycolysis and are sensitive to 2-deoxyglucose treatment, Cell Death
Dis. 5 (2014) e1336.
[132] Y.A. Shen, C.Y. Wang, Y.T. Hsieh, Y.J. Chen, Y.H. Wei, Metabolic reprogramming
orchestrates cancer stem cell properties in nasopharyngeal carcinoma, Cell Cycle
14 (2015) 86–98.
[133] W. Feng, A. Gentles, R.V. Nair, M. Huang, Y. Lin, C.Y. Lee, S. Cai, F.A. Scheeren,
A.H. Kuo, M. Diehn, Targeting unique metabolic properties of breast tumor
14
Biomedicine & Pharmacotherapy 112 (2019) 108690
initiating cells, Stem Cells 32 (2014) 1734–1745.
[134] G.J. Yoshida, H. Saya, Inversed relationship between CD44 variant and c-Myc due
to oxidative stress-induced canonical Wnt activation, Biochem. Biophys. Res.
Commun. 443 (2014) 622–627.
[135] G.J. Yoshida, Emerging roles of Myc in stem cell biology and novel tumor thera-
pies, J. Exp. Clin. Cancer Res. 37 (2018) 173.
[136] N. Wong, J. De Melo, D. Tang, PKM2, a central point of regulation in cancer
metabolism, Int. J. Biochem. Cell Biol. (2013).
[137] La Noce M, F. Paino, L. Mele, G. Papaccio, T. Regad, A. Lombardi, F. Papaccio,
V. Desiderio, V. Tirino, HDAC2 depletion promotes osteosarcoma’s stemness both
in vitro and in vivo: a study on a putative new target for CSCs directed therapy, J.
Exp. Clin. Cancer Res. 37 (2018) 296.
[138] E. Przybytkowski, D.A. Averill-Bates, Correlation between glutathione and sti-
mulation of the pentose phosphate cycle in situ in chinese Hamster ovary cells
exposed to hydrogen peroxide, Arch. Biochem. Biophys. 325 (1996) 91–98.
[139] S. Tuttle, T. Stamato, M.L. Perez, J. Biaglow, Glucose-6-phosphate dehydrogenase
and the oxidative pentose phosphate cycle protect cells against apoptosis induced
by low doses of ionizing radiation, Radiat. Res. 153 (2000) 781–787.
[140] G.C. Yeh, S.J. Occhipinti, K.H. Cowan, B.A. Chabner, C.E. Myers, Adriamycin re-
sistance in human tumor cells associated with marked alterations in the regulation
of the hexose monophosphate shunt and its response to oxidant stress, Cancer Res.
47 (1987) 5994–5999.
[141] C. Friesen, Y. Kiess, K. Debatin, A critical role of glutathione in determining
apoptosis sensitivity and resistance in leukemia cells, Cell Death Differ. 11
(2004) 73.
[142] T. Gessner, L.A. Vaughan, B.C. Beehler, C.J. Bartels, R.M. Baker, Elevated pentose
cycle and glucuronyltransferase in daunorubicin-resistant P388 cells, Cancer Res.
50 (1990) 3921–3927.
[143] L. Mele, F. Paino, F. Papaccio, T. Regad, D. Boocock, P. Stiuso, A. Lombardi,
D. Liccardo, G. Aquino, A. Barbieri, C. Arra, C. Coveney, M. La Noce, G. Papaccio,
M. Caraglia, V. Tirino, V. Desiderio, A new inhibitor of glucose-6-phosphate de-
hydrogenase blocks pentose phosphate pathway and suppresses malignant pro-
liferation and metastasis in vivo, Cell Death Dis. 9 (2018) 572.
[144] S.Y. Lee, E.K. Jeong, M.K. Ju, H.M. Jeon, M.Y. Kim, C.H. Kim, H.G. Park, S.I. Han,
H.S. Kang, Induction of metastasis, cancer stem cell phenotype, and oncogenic
metabolism in cancer cells by ionizing radiation, Mol. Cancer 16 (2017) 10.
[145] A. Mozumder, Y. Hatano, Charged Particle and Photon Interactions With Matter:
Chemical, Physicochemical, and Biological Consequences With Applications, 1 ed.,
CRC Press, 2003.
[146] J. Meesungnoen, J. Jay-Gerin, Radiation Chemistry of Liquid Water With Heavy
Ions: Monte Carlo Simulation Studies, CRC Press, Taylor & Francis, 2011, pp.
355–400.
[147] J.H. Tsai, J. Yang, Epithelial–mesenchymal plasticity in carcinoma metastasis,
Genes Dev. 27 (2013) 2192–2206.
[148] B. De Craene, G. Berx, Regulatory networks defining EMT during cancer initiation
and progression, Nat. Rev. Cancer 13 (2013) 97.
[149] S. Lamouille, J. Xu, R. Derynck, Molecular mechanisms of epithelial–mesenchymal
transition, Nat. Rev. Mol. Cell Biol. 15 (2014) 178.
[150] C. Lagadec, E. Vlashi, L. Della Donna, C. Dekmezian, F. Pajonk, Radiation‐induced
reprogramming of breast cancer cells, Stem Cells 30 (2012) 833–844.
[151] M. Diehn, R.W. Cho, N.A. Lobo, T. Kalisky, M.J. Dorie, A.N. Kulp, D. Qian,
J.S. Lam, L.E. Ailles, M. Wong, B. Joshua, M.J. Kaplan, I. Wapnir, F.M. Dirbas,
G. Somlo, C. Garberoglio, B. Paz, J. Shen, S.K. Lau, S.R. Quake, J.M. Brown,
I.L. Weissman, M.F. Clarke, Association of reactive oxygen species levels and
radioresistance in cancer stem cells, Nature 458 (2009) 780–783.
[152] H.Q. Duong, K.S. You, S. Oh, S.J. Kwak, Y.S. Seong, Silencing of NRF2 reduces the
expression of ALDH1A1 and ALDH3A1 and sensitizes to 5-FU in pancreatic cancer
cells, Antioxidants 6 (2017) 52.
[153] G.R. Buettner, One-electron and two-electron signaling in redox biology, Free
Radic. Biol. Med. 49 (2010) S7.
[154] M. Lopez-Lazaro, HIF-1: hypoxia-inducible factor or dysoxia-inducible factor?
FASEB J. 20 (2006) 828–832.
[155] R.D. Guzy, B. Hoyos, E. Robin, H. Chen, L. Liu, K.D. Mansfield, M.C. Simon,
U. Hammerling, P.T. Schumacker, Mitochondrial complex III is required for hy-
poxia-induced ROS production and cellular oxygen sensing, Cell Metab. 1 (2005)
401–408.
[156] K.D. Mansfield, R.D. Guzy, Y. Pan, R.M. Young, T.P. Cash, P.T. Schumacker,
M.C. Simon, Mitochondrial dysfunction resulting from loss of cytochrome c im-
pairs cellular oxygen sensing and hypoxic HIF-α activation, Cell Metab. 1 (2005)
393–399.
[157] Q. Cao, K.M. Mak, C. Ren, C.S. Lieber, Leptin stimulates tissue inhibitor of me-
talloproteinase-1 in human hepatic stellate cells respective roles of the JAK/STAT
and JAK-Mediated H2O2-Dependent MAPK pathways, J. Biol. Chem. 279 (2004)
4292–4304.
[158] D. Trachootham, W. Lu, M.A. Ogasawara, N.R.D. Valle, P. Huang, Redox regula-
tion of cell survival, Antioxid. Redox Signal. 10 (2008) 1343–1374.
[159] S. Kaewpila, S. Venkataraman, G.R. Buettner, L.W. Oberley, Manganese super-
oxide dismutase modulates hypoxia-inducible factor-1α induction via superoxide,
Cancer Res. 68 (2008) 2781–2788.
[160] Q. Liu, U. Berchner-Pfannschmidt, U. Moller, M. Brecht, C. Wotzlaw, H. Acker,
K. Jungermann, T. Kietzmann, A Fenton reaction at the endoplasmic reticulum is
involved in the redox control of hypoxia-inducible gene expression, Proc. Natl.
Acad. Sci. U. S. A. 101 (2004) 4302–4307.
[161] H. Shi, Hypoxia inducible factor 1 as a therapeutic target in ischemic stroke, Curr.
Med. Chem. 16 (2009) 4593–4600.
[162] S.J. Welsh, W.T. Bellamy, M.M. Briehl, G. Powis, The redox protein thioredoxin-1
Z. Ghanbari Movahed, et al.
(Trx-1) increases hypoxia-inducible factor 1α protein expression: Trx-1 over-
expression results in increased vascular endothelial growth factor production and
enhanced tumor angiogenesis, Cancer Res. 62 (2002) 5089–5095.
[163] S. Salceda, J. Caro, Hypoxia-inducible factor 1α (HIF-1α) protein is rapidly de-
graded by the ubiquitin-proteasome system under normoxic conditions its stabi-
lization by hypoxia depends on redox-induced changes, J. Biol. Chem. 272 (1997)
22642–22647.
[164] D.J. Lomb, L.A. Desouza, J.L. Franklin, R.S. Freeman, Prolyl hydroxylase inhibitors
depend on extracellular glucose and hypoxia-inducible factor (HIF)-2α to inhibit
cell death caused by nerve growth factor (NGF) deprivation: evidence that HIF-2α
has a role in NGF-promoted survival of sympathetic neurons, Mol. Pharmacol. 75
(2009) 1198–1209.
[165] T.L. Wellman, J. Jenkins, P.L. Penar, B. Tranmer, R. Zahr, K.M. Lounsbury, Nitric
oxide and reactive oxygen species exert opposing effects on the stability of hy-
poxia-inducible factor-1α (HIF-1α) in explants of human pial arteries, FASEB J. 18
(2004) 379–381.
[166] A. Gorlach, U. Berchner-Pfannschmidt, C. Wotzlaw, R.H. Cool, J. Fandrey,
H. Acker, K. Jungermann, T. Kietzmann, Reactive oxygen species modulate HIF-1
mediated PAI-1 expression: involvement of the GTPase Rac1, Thromb. Haemost.
89 (2003) 926–935.
[167] Q. Ding, E. Dimayuga, J.N. Keller, Proteasome regulation of oxidative stress in
aging and age-related diseases of the CNS, Antioxid. Redox Signal. 8 (2006)
163–172.
[168] A. Ciechanover, The ubiquitin proteolytic system from a vague idea, through basic
mechanisms, and onto human diseases and drug targeting, Neurology 66 (2006)
7–19.
[169] K.D. Wilkinson, Roles of ubiquitinylation in proteolysis and cellular regulation,
Annu. Rev. Nutr. 15 (1995) 161–189.
[170] C. Brahimi-Horn, J. Pouyssegur, When hypoxia signalling meets the ubiquitin-
proteasomal pathway, new targets for cancer therapy, Crit. Rev. Oncol. Hematol.
53 (2005) 115–123.
[171] R. Shringarpure, T. Grune, J. Mehlhase, K.J. Davies, Ubiquitin conjugation is not
required for the degradation of oxidized proteins by proteasome, J. Biol. Chem.
278 (2003) 311–318.
[172] T. Grune, T. Reinheckel, K.J. Davies, Degradation of oxidized proteins in K562
human hematopoietic cells by proteasome, J. Biol. Chem. 271 (1996)
15504–15509.
[173] T. Grune, T. Reinheckel, K. Davies, Degradation of oxidized proteins in mamma-
lian cells, FASEB J. 11 (1997) 526–534.
[174] T. Grune, T. Reinheckel, M. Joshi, K.J. Davies, Proteolysis in cultured liver epi-
thelial cells during oxidative stress Role of the multicatalytic proteinase complex,
proteasome, J. Biol. Chem. 270 (1995) 2344–2351.
[175] J.J. Haddad, Antioxidant and prooxidant mechanisms in the regulation of redox
(y)-sensitive transcription factors, Cell. Signal. 14 (2002) 879–897.
[176] M. Callapina, J. Zhou, T. Schmid, R. Kohl, B. Brune, NO restores HIF-1α hydro-
xylation during hypoxia: role of reactive oxygen species, Free Radic. Biol. Med. 39
(2005) 925–936.
[177] H. Elkon, E. Melamed, D. Offen, Oxidative stress, induced by 6-hydroxydopamine,
reduces proteasome activities in PC12 cells, J. Mol. Neurosci. 24 (2004) 387–400.
[178] T. Reinheckel, N. Sitte, O. Ullrich, U. Kuckelkorn, Kj. Davies, T. Grune,
Comparative resistance of the 20S and 26S proteasome to oxidative stress,
Biochem. J. 335 (1998) 637–642.
[179] O. Ullrich, T. Reinheckel, N. Sitte, R. Hass, T. Grune, K.J. Davies, Poly-ADP ribose
polymerase activates nuclear proteasome to degrade oxidatively damaged his-
tones, Proc. Natl. Acad. Sci. U. S. A. 6 (1999) 6223–6228.
[180] G.L. Wang, B.H. Jiang, G.L. Semenza, Effect of altered redox states on expression
and DNA-binding activity of hypoxia-inducible factor 1, Biochem. Biophys. Res.
Commun. 212 (1995) 550–556.
[181] B.M. Emerling, L.C. Platanias, E. Black, A.R. Nebreda, R.J. Davis, N.S. Chandel,
Mitochondrial reactive oxygen species activation of p38 mitogen-activated protein
kinase is required for hypoxia signaling, Mol. Cell. Biol. 25 (2005) 4853–4862.
[182] X. Kong, B. Alvarez-Castelao, Z. Lin, J.G. Castano, J. Caro, Constitutive/hypoxic
degradation of HIF-α proteins by the proteasome is independent of von Hippel
Lindau protein ubiquitylation and the transactivation activity of the protein, J.
Biol. Chem. 282 (2007) 15498–15505.
[183] M. Kaluzova, S. Kaluz, M.I. Lerman, E.J. Stanbridge, DNA damage is a prerequisite
for p53-mediated proteasomal degradation of HIF-1α in hypoxic cells and
15
Biomedicine & Pharmacotherapy 112 (2019) 108690
downregulation of the hypoxia marker carbonic anhydrase IX, Mol. Cell. Biol. 24
(2004) 5757–5766.
[184] M. Jahani, M. Azadbakht, F. Norooznezhad, K. Mansouri, L-arginine alters the
effect of 5-fluorouracil on breast cancer cells in favor of apoptosis, Biomed.
Pharmacother. 88 (2017) 114–123.
[185] Z.N. Demidenko, A. Rapisarda, M. Garayoa, P. Giannakakou, G. Melillo,
M.V. Blagosklonny, Accumulation of hypoxia-inducible factor-1α is limited by
transcription-dependent depletion, Oncogene 24 (2005) 4829.
[186] S. Chang, X. Jiang, C. Zhao, C. Lee, D.M. Ferriero, Exogenous low dose hydrogen
peroxide increases hypoxia-inducible factor-1alpha protein expression and induces
preconditioning protection against ischemia in primary cortical neurons, Neurosci.
Lett. 441 (2008) 134–138.
[187] E.L. Bell, T.A. Klimova, J. Eisenbart, C.T. Moraes, M.P. Murphy, G.S. Budinger,
N.S. Chandel, The Qo site of the mitochondrial complex III is required for the
transduction of hypoxic signaling via reactive oxygen species production, J. Cell
Biol. 177 (2007) 1029–1036.
[188] E.L. Bell, T.A. Klimova, J. Eisenbart, P.T. Schumacker, N.S. Chandel,
Mitochondrial reactive oxygen species trigger hypoxia-inducible factor-dependent
extension of the replicative life span during hypoxia, Mol. Cell. Biol. 27 (2007)
5737–5745.
[189] N.S. Chandel, D.S. McClintock, C.E. Feliciano, T.M. Wood, J.A. Melendez,
A.M. Rodriguez, P.T. Schumacker, Reactive oxygen species generated at mi-
tochondrial complex III stabilize hypoxia-inducible factor-1α during hypoxia a
mechanism of O2 sensing, J. Biol. Chem. 275 (2000) 25130–25138.
[190] R.D. Guzy, B. Sharma, E. Bell, N.S. Chandel, P.T. Schumacker, Loss of the SdhB,
but not the SdhA, subunit of complex II triggers reactive oxygen species-dependent
hypoxia-inducible factor activation and tumorigenesis, Mol. Cell. Biol. 28 (2008)
718–731.
[191] X. Lin, C.A. David, J.B. Donnelly, M. Michaelides, N.S. Chandel, X. Huang,
U. Warrior, F. Weinberg, K.V. Tormos, S.W. Fesik, A. Shen, Chemical genomics
screen highlights the essential role of mitochondria in HIF-1 regulation, Proc. Natl.
Acad. Sci. U. S. A. 105 (2008) 174–179.
[192] E.L. Page, D.A. Chan, A.J. Giaccia, M. Levine, D.E. Richard, Hypoxia-inducible
factor-1α stabilization in nonhypoxic conditions: role of oxidation and in-
tracellular ascorbate depletion, Mol. Biol. Cell 19 (2008) 86–94.
[193] V. Srinivas, I. Leshchinsky, N. Sang, M.P. King, A. Minchenko, J. Caro, Oxygen
sensing and HIF-1 activation does not require an active mitochondrial respiratory
chain electron-transfer pathway, J. Biol. Chem. 276 (2001) 21995–21998.
[194] M. Demaria, V. Poli, PKM2, STAT3 and HIF-1α: The Warburg’s vicious circle, Jak-
Stat. 1 (2012) 194–196.
[195] I. Schuppe‐Koistinen, P. Moldeus, T. Bergman, I.A. Cotgreave, S‐Thiolation of
human endothelial cell glyceraldehyde‐3‐phosphate dehydrogenase after hy-
drogen peroxide treatment, FEBS J. 221 (1994) 1033–1037.
[196] L. Cyrne, F. Antunes, A. Sousa-Lopes, J. Diaz-Berrio, H.S. Marinho,
Glyceraldehyde-3-phosphate dehydrogenase is largely unresponsive to low reg-
ulatory levels of hydrogen peroxide in Saccharomyces cerevisiae, BMC Biochem.
11 (2010) 49.
[197] L. Cao, J. Chen, M. Li, Y.Y. Qin, M. Sun, R. Sheng, F. Han, G. Wang, Z.H. Qin,
Endogenous level of TIGAR in brain is associated with vulnerability of neurons to
ischemic injury, Neurosci. Bull. 31 (2015) 527–540.
[198] J. Xiang, C. Wan, R. Guo, D. Guo, Is hydrogen peroxide a suitable apoptosis in-
ducer for all cell types? Biomed Res. Int. 2016 (2016) 1–6.
[199] M. Singh, H. Sharma, N. Singh, Hydrogen peroxide induces apoptosis in HeLa cells
through mitochondrial pathway, Mitochondrion 7 (2007) 367–373.
[200] O.D. Maddocks, C.F. Labuschagne, K.H. Vousden, Localization of NADPH pro-
duction: a wheel within a wheel, Mol. Cell 55 (2014) 158–160.
[201] R.J. DeBerardinis, N.S. Chandel, Fundamentals of cancer metabolism, Sci. Adv. 2
(2016) e1600200.
[202] D. Samanta, G.L. Semenza, Serine synthesis helps hypoxic cancer stem cells reg-
ulate redox, Cancer Res. 76 (2016) 6458–6462.
[203] D. Samanta, G.L. Semenza, Maintenance of redox homeostasis by hypoxia-in-
ducible factors, Redox Biol. 13 (2017) 331–335.
[204] S.M. Lee, T.L. Huh, J.W. Park, Inactivation of NADP+-dependent isocitrate de-
hydrogenase by reactive oxygen species, Biochimie 83 (2001) 1057–1065.
[205] K.M. Holmström, T. Finkel, Cellular mechanisms and physiological consequences
of redox-dependent signalling, Nat. Rev. Mol. Cell Biol. 15 (2014) 411–421.