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A mechanistic review of the anticancer potential of hesperidin, a

natural flavonoid from citrus fruits

Pratibha Pandey , Fahad Khan

PII: S0271-5317(21)00034-8
DOI: https://doi.org/10.1016/j.nutres.2021.05.011
Reference: NTR 8241

To appear in: Nutrition Research

Received date: 2 March 2021

Revised date: 23 May 2021
Accepted date: 31 May 2021

Please cite this article as: Pratibha Pandey , Fahad Khan , A mechanistic review of the anti-
cancer potential of hesperidin, a natural flavonoid from citrus fruits, Nutrition Research (2021), doi:
https://doi.org/10.1016/j.nutres.2021.05.011

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 Highlights

 Perspective of hesperidin as potential anticancer drug candidate

 Therapeutic potential of hesperidin in in vivo and in vitro models

 Major targets of hesperidin and their associated signaling pathways

 Enhanced anticancerous properties of hesperidin formulations

 A mechanistic review of the anticancer potential of hesperidin, a natural flavonoid from

citrus fruits

Pratibha Pandey1, Fahad Khan1*

1
Department of Biotechnology, Noida Institute of Engineering & Technology, 19, Knowledge

Park-II, Institutional Area, Greater Noida, 201306, India

*Corresponding author:
Dr. Fahad Khan
Assistant Professor
Department of Biotechnology
Noida Institute of Engineering and Technology, Greater Noida
fahadintegralian@gmail.com
Abstract

Hesperidin, a phytoactive compound, is an abundant and economical dietary bioflavonoid

possessing numerous biological and medicinal benefits. Several studies have strongly proven the

significant chemotherapeutic potential of hesperidin. Therefore this review is an effort to bring

together the existing studies demonstrating hesperidin as a potential anticancer agent with its

mode of action reported in the therapeutic strategies for numerous cancer types. Hesperidin acts

via modulating multiple pathways involving cell cycle arrest, apoptosis, antiangiogenic,

antimetastatic and DNA repair in various cancer cells. Hesperidin has been reported to alter

several molecular targets related to carcinogenesis, such as reactive nitrogen species, cellular

kinases, transcription factors, reactive oxygen species, drug transporters, cell cycle mediators and

inflammatory cytokines. Altogether, this review provides significant insights for the potential of

hesperidin to be a strong and promising candidate for pharmaceuticals, functional foods, dietary

supplements, nutraceuticals and geared toward the better management of carcinoma.

Keywords

Anticancer potential; Phytocompound; Metabolism; Hesperidin; Citrus

1. Introduction

Bioflavonoids have been gaining wider attention in cancer management for their strong

antioxidant and bioactive potential (1). Hesperidin (3,5,7-trihydroxyflavanone-7-

rhamnoglucoside) is a natural phenolic compound with several biological effects (2). It is a much

known fact that products derived from citrus fruits have been widely used for combating several

human ailments for thousands of years (3). Flavonoids are considered to be the major category of

such dietary polyphenols (4). Amongst diverse flavonoids, hesperidin, a flavanone glycoside, is

widely found in sweet oranges, lemons, and some other vegetables and fruits (5). It is a colorless

and odorless plant chemical with the chemical formula of C28H34O15 and a molecular weight of

610.1898 g/mol (Figure 1A). Its name was derived from the word "hesperidium", specifically for

the fruit of citrus trees (6). Various studies have reported limited bioavailability and poor

absorption of hesperidin in experimental animals due to the attachment of rutinoside moiety with

the dietary flavonoid (7). However, hesperidin has been recognized as a potent candidate

phytocompound with numerous health benefits such as antimicrobial, anticancer, antioxidant,

anti-inflammatory, antidiabetic, and cardiovascular protective activities (8). Specifically,

hesperidin has been confirmed to possess both chemopreventive and chemotherapeutic potential

(9). To illustrate a better perceptive of the anticancerous potential of hesperidin, this review

focuses on major natural sources of hesperidin, its bioavailability, anticancer potential,

pharmacokinetics, and its associated molecular mechanisms. Furthermore, the anticancer

potential of hesperidin analogs, hesperidin-nanoformulations, and hesperidin metabolites are also

outlined. Several safety issues associated with the usage of hesperidin as a dietary component are

also summarized. We emphasize that hesperidin is a promising natural flavonoid with significant

anticancerous potential, and this review would be highly helpful in furthering the usage of
hesperidin as one of the natural compounds for the prevention and management of various

carcinomas.

2. Natural sources of hesperidin

Hesperidin is isolated from ordinary orange Citrus aurantium L, C. sinensis, C. unshiu, and other

species of the genus Citrus (10). It has also been reported in several other plants (apart from

Citrus) such as in Betulaceae, Fabaceae, Papilionaceae and Lamiaceae, the bark of Zanthoxylum.

cuspidatum, and Zanthoxylum avicennae. Hesperidin is found in various vegetables, fruits, and

several medicinal plants. Though hesperidin is widely distributed in many plants, Cyclopia

maculata stems have been recognized as the major source of hesperidin (11). Citrus fruits,

Cyclopia maculate tea, and oranges are also considered as rich sources of hesperidin. Table 1

outlines the approximate hesperidin content of selected plants with medicinal and nutritional

value.

3. Absorption and metabolism of hesperidin

Numerous research reports have emphasized the importance of absorption and metabolism of

phytocompound for its further utilization as the safe and effective anticancer lead candidate (19).

Dietary flavonoids usually have limited or low bioavailability. During their oral administration,

hesperidin is converted to hesperetin (a trihydroxyflavanone), an aglycone of hesperidin, by β-

glucosidase present in intestinal microflora and then absorbed from the gastrointestinal tract

(GIT) (20). Hesperetin is the predominant flavonoid with having three hydroxy groups at 3'-, 5-

and 7-positions and methoxy substituent at the 4'-position (Figure 1B). Additionally, several

reports have displayed the degradation of flavonoid into several carboxylic acid and phenolic

products after the flavonoid glycosides hydrolysis by gut bacteria (21). Thus, intestinal bacteria

metabolism might play an important role in hesperidin uptake. It is therefore conceivable that
changes in intestinal microflora might affect the absorption and subsequent pharmacokinetics of

hesperidin, and consequently lead to alterations in the biological activity exhibited by hesperidin.

The gut microbiome exhibits a significant role in hesperidin metabolism (22). Several studies

had reported that intake of citrus juice leads to the suppression of inflammatory enzymes and

pro-inflammatory cytokines expression in the colon, thereby resulting in the inhibition of colon

cancer (23), and further explained that some intestinal bacterial enzymes such as rhamnosidases

may exhibit differentiated activity between neohesperidoside moiety of naringenin and rutinoside

moiety of hesperetin (hesperidin). Hesperidin might increase the epithelial barrier role of Caco-2

cells in a concentration-dependent manner (24). Three types of metabolites of hesperidin

including sulfates, aglycons, phenolic acids were absorbed in the intestinal tissue. The liver and

intestinal tract are the key metabolic site, where both host metabolic enzymes and intestinal

bacteria take part in chemical biotransformation (25). Hesperidin is known to be mostly resistant

to enzymatic breakdown in the small intestine and stomach, thus, mainly get into the intact colon

(26). Several earlier reports have suggested the appearance of hesperidin in the small intestine,

even though the majority is transformed in the colon, some breakdown can previously occur in

the small intestine (distal part) (27). It was also found that the antiproliferative and antioxidant

activities of flavonoids were greatly improved after its deglycosylation (28-29). Thus, after

administration of hesperetin in the duodenum, cecum, and colon were valuable to several

intestinal bioactivities. Several bacteria are found in the intestinal tract and gut microbiota

contains numerous enzymes which modify food compounds (30). Microbial enzymes can

remove glucuronides, glycosides, sulfates, and produce glycons (flavonoid), which gets further

metabolized into several ring fission products, thus, a range of phenolic acid metabolites was

produced after hesperidin administration in mice, these metabolites were absorbed in the colonic
tissue (32). Previously reported studies suggested that the metabolites exhibited decreased

polarity, smaller molecular weight, and stronger retention in comparison to the prototype that

might improve absorption and distribution during in vivo (33). Both phase II enzymes including

UGT (uridine-5ʹ-diphosphate-glucuronosyl transferases) and SULT (sulfotransferases) are

associated with the production of the conjugated metabolites (34). Both enzymes are present in

various tissues such as the small intestine, liver, and colon (35). These experiments further

indicated that the sulfates of hesperidin absorbed in the ileum, after oral administration of

hesperidin, the conjugated metabolites would be absorbed in the ileum and duodenum.

Hesperidin metabolite production is commonly contributed by gut microbiota, liver, and

enterocyte (36). Altogether, hesperidin and its metabolites have been broadly absorbed in various

intestinal tissues. After oral consumption of hesperidin, conjugates of glucuronide, and sulfate

(hesperetin) were found in plasma demonstrating absorption in the small intestine. The

microbiota could produce rhamnosidases that perform absorption by cleaving the rhamnose

moiety from hesperidin.

4. Anticancerous effects of hesperidin

4.1 In vitro anticancerous efficacy

Hesperidin has been reported to limit the proliferation of several cancer cells in vitro, such as

glioma, prostate, pancreas, ovarian, skin, liver, colon, lung, cervical, and breast cancer cells (37).

Table 2 summarizes the recent reports on the anticancerous efficacy of hesperidin and its

associated molecular mechanism behind their mode of action in various cancer cells. Hesperidin

concentration varies with the origin of different cancer cell types and showed significant

anticancerous potential at relatively high doses in in vitro studies (38-39). Hesperidin contributes

multifaceted roles in anticancerous potential, such as inhibition of the invasion, proliferation,

metastasis, migration, and angiogenesis of cancer cells, along with apoptotic and autophagic

induction. Anticancerous efficacy of hesperidin is mediated via the regulation of numerous

molecular signaling pathways and molecular targets involved in carcinogenesis. Altogether,

hesperidin is a promising anticancer phytocompound against several cancer cells (in vitro) (40).

Phytocompounds can increase the cytotoxicity of various chemotherapeutic drugs and irradiation

by decreasing the drug-induced toxicity as well as increasing the sensitivity of several cancer

cells. Hesperidin either alone or in combination with chemotherapeutic drugs, including

paclitaxel, tamoxifen, cyclophosphamide, temozolomide, pirarubicin, methotrexate, oxaliplatin,

and doxorubicin has been reported with significant tumor growth inhibition (Table 3). A

combination of hesperidin and naringenin, a bioactive flavonoid exhibited synergistic

anticancerous effects in pancreatic cancer cells (41). Synergistic anticancerous effects of

hesperidin with chemotherapeutic drugs or irradiation are mediated via apoptotic induction, drug

efflux pumps inhibition, modulation of several cell signaling pathways reported in cancer

progression and, cell cycle protein regulation (Table 3). Altogether, these in vitro studies suggest

that hesperidin could be a potential anticancerous adjuvant agent.

4.2 In vivo anticancerous efficacy

In vivo anticancerous efficacy of hesperidin has been explained in xenograft mice models (58).

Hesperidin has exhibited significant anticancerous potential against numerous cancer types in

vivo, including prostate, breast, colon, and lymphoma. Swiss albino mice, Balb/c nude mice, and

Wistar rat models are commonly used to investigate the chemotherapeutic potential of hesperidin

(59). Cancer models are established by implantation of xenografts or murine cancer cells via

intraperitoneal or subcutaneous injection. Several studies have reported the hesperidin

administration either by oral or intraperitoneal administration at varying doses and different

treatment duration depending on the experimental setup. The mechanism behind the tumor

growth inhibitory potential of hesperidin in xenograft has been mainly associated with apoptotic

induction, angiogenesis inhibition, metastasis suppression, and regulation of associated cell

signaling pathways (60). Table 4 summarizes the in vivo anticancerous potential of hesperidin.

5. Anticancer mechanism of hesperidin

Several research studies have reported the role of hesperidin in cell cycle progression arrest in

various cancer cells (67). Hesperidin can arrest cell cycle progression at different checkpoints

including G1, G2/M, or S phase (68). Hesperidin-mediated cell cycle arrest may be associated

with the modulation of cell cycle regulatory proteins including CDK inhibitors, cyclin-dependant

kinases, and cyclins (Figure 2). Hesperidin was reported to enhance wild-type p53 levels in lung

cancer, breast cancer, leukemia cell lines, and colon cancer (69). Xia et al., 2018 reported that

hesperidin inhibited the G1/S transition in the A549 lung adenocarcinoma cell line via

upregulated p21 and downregulated cyclin D1 expression levels. Altogether, hesperidin can

target cell cycle signaling pathways in the cancer therapeutic approach. WNT/β-catenin signaling

pathway contributes a major role in cell proliferation, growth, and survival (70). Aberrant

WNT/β-catenin signaling pathway has been associated with numerous carcinomas such as

prostate cancer, colorectal cancer, and breast cancer (71). Thus, β-catenin signaling has been

considered as one of the attractive targets in cancer therapeutics (72). Hesperidin has been

reported to induce apoptosis in lung cancer cells via downregulation of PCNA protein, β-catenin,

and c-myc, expressions. Wei Hong et al., 2020 reported the therapeutic potential of hesperidin in

promoting the differentiation of human alveolar osteoblasts through the Wnt/β-Catenin signaling

pathway activation. Hesperidin has also presented significant hepatoprotective efficacy in the
hepatocellular carcinoma group via increased β-catenin, Wnt3a, Wnt5a, Cyclin D1 gene, and

protein expressions (73).

Hesperidin has depicted significant anticancerous potential via targeting apoptosis-mediated cell

death pathways. Recent studies have reported apoptosis as one of the major phenomena by which

various anticancerous agents remove neoplastic or preneoplastic cells (74, 75). Through

literature findings, it was postulated that hesperidin induces apoptosis via several mechanisms

such as increasing DNA fragmentation and nuclear condensation. These apoptosis-inducing

effects of hesperidin are associated with caspase-9 and -3 activation, inhibition of cell cycle

progression, regulation of Bcl-2 (B-cell lymphoma-2) family proteins, decreased levels of NF-κB

(nuclear factor-κB), and elevation of reactive oxygen species levels. Hesperidin was also

reported with inhibition of MMP-9, COX-2 (cyclooxygenase-2), MMP-2 (matrix

metalloproteinase-2), and regulation of mitogen-activated protein kinases (c-Jun N-terminal

kinases and extracellular signal-regulated kinases) phosphorylation (76). Hesperidin induced

apoptosis through triggering of mitochondrial apoptotic pathway and inducing G0/G1 arrest in

A549 (human non-small cell lung cancer) cells (77). Hesperidin also promoted programmed cell

death via downregulation of estrogen receptor signaling (non-genomic) pathway in endometrial

cancer cells (78). Besides, hesperidin activated both intrinsic and extrinsic pathways of apoptosis

in several cancer cells via downregulation of Bcl-2 and upregulation of caspases (caspase-3, 8,9),

and Bax (79). This research strongly supported the fact that hesperidin can induce both intrinsic

(mitochondria-mediated) and extrinsic (death receptor-mediated) apoptotic pathways in various

cancer cells. p53 (tumor suppressor gene) activation is correlated with apoptotic induction via

negative and positive regulation of Bcl-2 and Bax (80). The role of hesperidin in apoptotic

induction has also been confirmed via p53 activation (81). Hesperidin inhibited
hepatocarcinogenesis and suppressed cell proliferation, inflammation, collagen deposition,

oxidative stress via activation of PPARγ and Nrf2/ HO-1/ARE pathways (82).

ROS (reactive oxygen species) formations during metabolism have been involved in various

physiological functions (83). The balance between ROS production and its elimination via

antioxidant defense system has been properly maintained in normal cells whereas ROS

homeostasis is deregulated in cancer cells, leading to higher ROS generation (84). Nonetheless,

if ROS generation reaches beyond its threshold level, it could trigger apoptotic induction in

cancer cells, thereby limiting cancer progression, explaining that ROS can be identified as one of

the potent therapeutic tools in cancer management (85). Several studies have reported that

flavonoids induce anticancer potential via increased ROS generation (86). For instance, the

anticancerous potential of hesperidin is regulated by ROS-dependent apoptotic pathways in

several cancer cells such as primary gall bladder cancer cells, HeLa, HepG2, and prostate cancer

cells (87). Also, hesperidin can be a good ROS scavenger and could act as a potent antioxidant

(non-enzymatic) defense system. Hesperidin has also been reported to promote the loss of MMP

(mitochondrial membrane potential), ROS formation, intracellular Ca2+ mobilization, caspase-3

activation, enhanced release of apoptosis-inducing factor, and cytochrome c from mitochondria

(87). It also mediated cell cycle growth arrest in the G0/G1 phase in HeLa cells via

downregulation of cyclinE1, cyclinD1, and CDKs (cyclin-dependent kinase 2) protein expression

level. In this perspective, hesperidin, induced apoptosis in human HepG2 cells via death receptor

pathway and mitochondrial pathway (88). Hesperidin administration has also shown a significant

reduction in the level of serum AFP level, liver function enzymes, and oxidative stress marker.

Various cellular kinases are recognized as the molecular targets of hesperidin (89). EGFR

(receptor tyrosine kinase), is also activated by its ligands that further promote the
phosphorylation of PI3K (phosphoinositide 3-kinase) and MAPK signaling, contributing a

prominent role in cell survival (90). Furthermore, hesperidin treatment suppressed DEN-induced

upregulation of CDK‐2, PI3K, Akt protein expression, and maintained the integrity of liver

tissues from hepatocellular carcinoma formation (91-92). Altogether, the hepatoprotective

potential of hesperidin is mediated through downregulation of the PI3K/Akt pathway.

Inflammation has been considered as one of the crucial contributing factors in cancer

progression. Pro-inflammatory molecules, including interferon c, interleukins, and TNFα, have

also been implicated in cancer progression and development. During inflammation, MAPK

pathway modulation results in activation of AP-1 and NF- ⱪB which is further associated with

increased COX-2 and iNOS gene expression (93-94). Hesperidin can stimulate TNFα secretion

in numerous cancer cells, including A549 and LAN-5 cells (95). Overall, hesperidin has

exhibited significant anticancerous potential through the suppression of the production of various

inflammatory molecules involved in cancer development and progression (Figure 2).

6. Anticancer efficacy of hesperidin analogs, nanoformulations, and metabolites

The anticancerous properties of bioactive phytocompounds can be significantly improved by

various nanoparticle formulations as well as enzymatic and chemical modifications (96). Due to

its poor bioavailability and low solubility, access of hesperidin into cells for its pharmacological

properties remains limited. Numerous drug delivery approaches, including microencapsulation,

enzymatic modification, and complexation have been recommended for enhancing its targeted

delivery, solubility and bioavailability thereby improving its anticancer, and biological activity

(97). According to Ali et al., 2019 (98), modified nanohesperidin exhibited higher growth

inhibitory potential and better cytocompatibility than the native hesperidin in breast cancer cells

by modulating the apoptotic pathway of Caspase-3 and p53. Research conducted by Byun et al.,
2019 explained that hesperidin irradiated with 150 kGy gamma has also resulted in higher

growth inhibition of B16BL6 melanoma cells and lung metastasis in C57BL/6 mice in

comparison to non-irradiated hesperidin (99). Balakrishnan et al., 2020 (100) reported that

formulated hesperidin nanoparticles were more competent in comparison to native hesperidin in

Hep-2 cells. Altogether, hesperidin could be considered a safe and economical drug candidate for

the better management of cancer treatments.

7. Conclusion

Increasing cancer incidence, costlier anticancer drugs with their severe side effects, multidrug

resistance are some of the major challenges in cancer therapeutics. Furthermore, various

chemotherapeutic drugs fail to manage numerous carcinomas. There has been mounting demand

for phytocompounds as potent anticancer drugs because they are easily available with low or

minimal toxicity. It is evident from our review that hesperidin could be a promising anti-cancer

agent. Combining chemotherapeutic drugs with phytocompounds would be a better approach for

the treatment of cancer. In combination, hesperidin usually does not affect the activity of chemo

drugs, but somewhat increases the drug-induced cytotoxicity in organ tissues. Hesperidin has

exhibited both chemopreventive and chemotherapeutic effects against numerous carcinomas.

Hesperidin could affect several cell signaling pathways associated with cancer progression.

Several approaches including the combination of hesperidin with other phytochemicals,

chemotherapeutic drugs, irradiation, and nanoformulations could be utilized further to improve

its bioavailability and efficacy as an anticancer agent. Nevertheless, the antitumor efficacy of

hesperidin has been elucidated only in preclinical studies including in vivo and in vitro cancer

models. However respect to limited clinical studies related to pharmacological properties of

hesperidin, it is quite difficult to draw a clear picture about the effective dose consideration for
cancer management in the human body. Therefore, further studies are still needed to elucidate

the best effective doses for future clinical trials in cancer patients to validate the candidature of

hesperidin as one of the promising and effective therapeutic strategies for cancer management.

Sources of Support

This research received no external funding

Declaration of competing interest

All authors declare no conflict of interest.

Acknowledgements

The authors thank the management of Noida Institute of Engineering & Technology for

providing an opportunity to complete the review.

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Figure 1: Chemical structure of (A) Hesperidin and (B) Hesperitin. PubChem database has been

used to retrieve the chemical structure of these two potential phytocompounds.

Figure 2: Mechanism associated with the anticancer potential of hesperidin and its associated

targets
Table 1: Occurrence of hesperidin in plants

Plant species Hesperidin content References

Citrus limonia 15.4 g kg-1 (12)
Grape fruits (Citrus paradisi) 11.15 % w/w
Mosambi (Citrus limetta) 3.79% w/w
(13)
Lemon (Citrus lemon) 1.72% w/w
Orange (Citrus sinensis) 1.29% w/w
Croatian mandarin peels 31.42 mg/g (14)
Mentha piperita L. 504.2 mg/L (15)
Stevia rebaudiana 493.4 mg/L
Sorbus tianschanica leaves 0.48 mg/g (16)
Citrus aurantium var. bigaradia 2.66 (w/w) % (17)
0.62 mg/g (bagasse)
Citrus limetta 27.3 mg/L (juice) (18)
0.04 mg/g (seed)
0.98 mg/g (bagasse)
Citrus reticulate 21.7 mg/L (juice) (18)
0.09 mg/g (seed)
Citrus sinensis L 23.0 µg/mL (19)
Table 2: In vitro studies of anticancerous effects of hesperidin

Hesperidin
Cancer cell lines Anticancerous potential References
concentration
Maximizes the anticancerous effects in
human pancreatic cancer via down
Panc-1 cells 1-20µM (42)
regulation of p38 and FAK signaling
pathways
Significant apoptotic cells accumulation in
G0/G1 cell cycle phases, apoptotic
MCF-7 human
induction via extrinsic and intrinsic
breast cancer cell 20-40µg/mL (43)
pathways, down regulation of anti
line
apoptotic gene (Bcl-2), and upregulation of
pro-apoptotic gene (Bax).
Decreased PD-L1 expression via inhibition
MDA-MB231
of NF-κB and PI3K/Akt pathway.
triple negative
Hesperidin prevented metastasis
breast 10 to 50 µM (44)
phenotypes via reduction in PD-L1
adenocarcinoma
(immune checkpoint inhibitor) expression
cancer cell line
PD-L1
Hesperidin administration resulted in
alleviation of non small cell lung cancer
A549 and H460
1-2.5 µM by antiproliferative effect and apoptotic (45)
cells
induction in NSCLC cells via the miR-
132/ZEB2 pathway.
Hesperidin loaded nanocomposites
administration induced apoptosis via
HCT116 cells 5-15 µg/mL enhanced ROS generation leading to (46)
excessive mitochondrial membrane
permeability in HCT116 cancer cells.
Apoptosis inducing potential via enhanced
reactive oxygen species generation, loss of
MMP (mitochondrial membrane potential)
Human gall
25-200 µM cell cycle arrest, and caspase-3 activation (47)
bladder carcinoma
in the primary cells generated from
surgically removed cancerous gall bladder
tissues.
Hesperidin inhibits proliferation through
apoptosis mediated by endoplasmic
HeLa cell 20-100 µM (47)
reticulum stress pathways and cell cycle
arrest
Hesperidin treatment induces paraptosis
HepG2 Cells 0.1-2 mM like cell death via phosphorylation of (48)
ERK1/2
 Reduces cell viability, and induces
apoptosis via activated protein expression
A2780 cells 0.1-10 µM (49)
of caspase-3 through cytochrome c
signaling pathways
Inhibition of cell proliferation via down
regulation of β-catenin, PCNA, and c-myc
protein expression. apoptotic induction via
A549 cells 6.25-100 µM (50)
enhanced levels of APAF-1, cytochrome c,
DNA fragmentation, and caspase-3 in
hesperidin treated cells
Exhibited cytotoxic, anti-proliferative and
HT-29 and HCT-
58.45 μmol/L pro-apoptotic effect via AMPK/mTOR (51)
116
signaling pathway
Induces mitochondrial dependent apoptosis
Gastric cancer
1-100 μM by increasing ROS levels and MAPK (51)
(GC) AGS cells
signaling pathway regulation in AGS cells
Abbreviations: FAK, focal adhesion kinase; PD-L1, programmed death-ligand 1; NF-κB,
nuclear factor kappa light chain enhancer of activated B cells; PI3K, phosphatidylinositol 3-
kinase; Akt, protein kinase B; NSCLC, non-small-cell lung carcinoma; ZEB2, zinc finger E-box
Binding homeobox 2; ROS, reactive oxygen species; MMP, mitochondrial membrane potential;
ERK1, extracellular-signal-regulated kinase PCNA, proliferating cell nuclear antigen; c-myc,
cellular myelocytomatosis oncogene; AMPK, AMP-activated protein kinase; mTOR, mammalian
target of rapamycin complex 1; AGS, adenocarcinoma gastric cell line

Table 3: Studies on anticancerous potential of combinations of hesperidin and

chemotherapeutic drug/irradiation

Cancer
Combinatorial treatment Synergistic effects References
cells/Model
Hesperidin in combination with
gemicitabine synergistically
Gall bladder
Hesperidin+Gemicitabine reduced the proliferation of GBC (47, 52)
carcinoma
cells in a more significant way in
comparison to hesperidin alone
Combinatorial treatment of
MCF–7 breast
hesperidin and doxorubicin
cancer cells Hesperidin+Doxorubicin (53)
resulted in the inhibition of Pgp
line
expression in MCF-7/Dox cells
Combined treatment of
hesperetin and platinum
exhibited more significant
A549 cells Hesperetin + Platinum (54)
inhibitory effect on lung cancer
progression in compared with
single drug treatment.
 Combined treatment induced
4T1 murine apoptosis and cell cycle arrest in
metastatic G2/M phase. Combined
Hesperitin+Doxorubicin (55)
breast cancer treatment inhibited MMP-9
cells expression and migration and in
4T1 cells
Synergistically enhance the
anticancerous effect of
MCF7 and Tamoxifen+Hesperidin+Pip
tamoxifen and can be further (56)
T47D cells erine
utilized as safe adjuvant or
vehicle to tamoxifen
Rat model
Significant increase in apoptotic
(Sprague-
effects induced by 5-FU via
Dawley) of Hesperetin+5-fluorouracil
remarkable elevation in gene (57)
HCC (5-FU)
expression of FasL, Fas,
(hepatocellular
caspase-3 and caspase-8
carcinoma)
Abbreviations: GBC, gall bladder cancer; MCF-7, michigan cancer foundation-7; Dox,
Doxorubicin; FasL, fas-ligand; MMP-9, matrix metallopeptidase 9; T47D, Human Breast Cancer
Cells; 4T1, breast cancer cell line derived from the mammary gland tissue of a mouse BALB/c
strain; Pgp, P-glycoprotein
Table 4: In vivo anticancerous studies of Hesperidin

Animal cancer Hesperidin Outcome and possible mode of

References
BALB/c mice model treatment action
Apoptosis induction in tumor cells
Ehrlich ascites
100 mg/kg via DNA fragmentation, down-
carcinoma (EAC) (60)
regulation of antiapoptotic gene
bearing Balb/C mice
Bcl2 and Caspase 3 stimulation
Displayed in vivo and in vitro
apoptotic and antitumor effects in
Bone cancer
MG-63 cells via mediating cell
MG-63 xenograft/ 80 mg/kg (61)
cycle arrest, cell migration
BALB/c mice
inhibition and mitochondrial
membrane mediated apoptosis.
Inhibited esophageal cancer cell
Esophageal cancer
growth and invasion by inhibiting
Eca109 xenograft/ 90 mg/kg (62)
or downregulating the PI3K/AKT
BALB/c-nu/nu mice
signaling pathway.
Cell apoptosis induction in
Esophageal cancer
esophageal cancer cells via
Eca109 xenograft/ 60 mg/kg (63)
mitochondrial mediated intrinsic
BALB/c-nu/nu mice
pathway by ROS accumulation.
hesperidin to be a potent
chemopreventive agent against
DEN-induced Kidney renal carcinogenesis possibly by
200 mg/kg (64)
Cancer/ Wistar rats virtue of its antioxidant properties
and by modulation of multiple
molecular pathways.
Hesperidin treatments combat with
AOM-induced the inflammation via
Colorectal cancer/ 25 mg/kg downregulation of NF-κB and its (65)
Swiss albino mice related target molecules COX-2 and
iNOS.
Cyclophosphamide
Hesperidin treatment resulted in an
induced Premature
80 mg/kg effective treatment for premature (66)
ovarian failure (POF)/
ovarian failure in rats.
rat model
Abbreviations: Bcl-2, B-cell lymphoma 2; BALB, bagg albino; COX-2, cyclooxygenase-2;
iNOS, inducible nitric oxide synthase; DEN, diethylnitrosamine; AOM, azoxymethane; NF-κB,
nuclear factor kappa light chain enhancer of activated B cells; ROS, reactive oxygen species
Figure 1.
Figure 2.
Graphical abstract