Cancer Cell
Previews
ROS Promotes Cancer Cell Survival
through Calcium Signaling
Colleen R. Reczek1 and Navdeep S. Chandel1,*
1Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA
*Correspondence: nav@northwestern.edu
https://doi.org/10.1016/j.ccell.2018.05.010
To avoid reactive oxygen species (ROS)-induced cell death, cancer cells increase their antioxidant defense
system. In this issue of Cancer Cell, Takahashi et al. identify a novel, non-canonical oxidative stress defense
mechanism involving TRPA1, a redox-sensitive Ca2+ channel, and the upregulation of anti-apoptotic path-
ways to promote cancer cell survival.
Cancer cells rely on the signaling capa-
bilities of reactive oxygen species
(ROS) for cell migration, proliferation,
and survival. To increase ROS produc-
tion, cancer cells acquire oncogenic
mutations, lose tumor suppressors, and
accelerate their metabolism. Hydrogen
peroxide (H2O2), a type of ROS, pro-
motes pro-tumorigenic signaling by
mediating the cysteine oxidation of
target proteins to alter their function.
Importantly, if ROS levels are too high,
ROS can promote anti-tumorigenic
signaling and induce cancer cell senes-
cence and cell death (Reczek and Chan-
del, 2017). Therefore, it is critical for
cancer cells to maintain an optimal
ROS level. To prevent excessive ROS
accumulation, cancer cells increase their
antioxidant capacity (Gorrini et al.,
2013). This canonical ROS-neutralizing
mechanism includes upregulation of the
redox-sensitive transcription factor nu-
clear factor erythroid 2-related factor 2
(NRF2). NRF2 protein stabilization
during oxidative stress allows for the
induction of numerous antioxidants,
including enzymes involved in gluta-
thione (GSH) synthesis and utilization,
glutathione peroxidase, and peroxire-
doxin (Gorrini et al., 2013). Given that
cancer cells require ROS homeostasis,
anti-cancer therapies that manipulate
ROS levels seem attractive. Inhibiting
key antioxidant enzymes may elevate
ROS levels to promote oxidative stress-
induced cancer cell death. However, it
is important to consider with this
approach that normal, non-transformed
cells are also sensitive to high ROS
levels; therefore, a viable therapeutic
strategy should involve selectively tar-
geting cancer cells.
In this issue of Cancer Cell, Takahashi
et al. report an unconventional oxidative
stress defense mechanism whereby tran-
sient receptor potential ankyrin 1 (TRPA1)
mediates the upregulation of calcium
(Ca2+)-dependent anti-apoptotic path-
ways to prevent ROS-induced cancer
cell death (Takahashi et al., 2018).
TRPA1 is one member of the TRP family
of ion channel proteins. It is a plasma
membrane-bound protein well known
for its chemosensing ability, mediating
neurogenic pain and inflammation (Paul-
sen et al., 2015). TRPA1 has been shown
to be activated by numerous noxious
compounds, including H2O2, which oxi-
dizes hyper-reactive cysteine residues
on its cytoplasmic side (Bessac et al.,
2008; Paulsen et al., 2015). Activation of
TRPA1 leads to the influx of Ca2+, which
can serve as an important signaling mole-
cule to regulate numerous physiological
and biological responses, including cell
proliferation and survival (Bessac et al.,
2008; Déliot and Constantin, 2015; Paul-
sen et al., 2015). Importantly, these
Ca2+-dependent cellular responses are
also implicated in tumorigenesis, sug-
gesting a role for TRPA1 in cancer. Previ-
ously, the antibacterial agent triclosan
was shown to induce TRPA1 activation
and Ca2+ influx in human prostate cancer
stromal cells, resulting in vascular endo-
thelial growth factor (VEGF) secretion
and increased prostate cancer epithelial
cell growth (Derouiche et al., 2017).
Moreover, through its interaction with
FGFR2, TRPA1 was shown to support
lung adenocarcinoma cell proliferation
and metastasis (Berrout et al., 2017).
Although TRPA1 has been implicated in
cancer, its redox-sensing capabilities
and potential effect on oxidative stress
Cancer Cell 33, June 11, 2018 ª 2018 Elsevier Inc. 949
defense programs have not been
examined.
Takahashi et al. (2018) systematically
determined whether and how ROS-
induced TRPA1 activation and Ca2+ influx
affect the oxidative stress defense pro-
grams in cancer cells to promote cancer
cell survival. The authors first examined
whether TRPA1 was upregulated and
functional in a variety of cancer types;
this would be expected if cancer cells
use TRPA1 to tolerate oxidative stress.
Indeed, Takahashi et al. found increased
TRPA1 mRNA and protein expression in
breast and lung tumors. Moreover, by us-
ing the TRPA1 agonist mustard oil in
combination with genetic (TRPA1 knock-
down) and pharmacological (AP-18, a
TRPA1-specific inhibitor) approaches,
the authors showed that TRPA1 medi-
ates an increase in the intracellular Ca2+
concentration in numerous cancer cell
lines. Next, the authors showed that a
TRPA1-mediated Ca2+ influx was
required for the increased survival of
TRPA1-enriched cancer cells in response
to H2O2. Furthermore, ectopic expres-
sion of TRPA1 was sufficient to improve
the cell survival of TRPA1-low-express-
ing cancer cells. Using a more physiolog-
ically relevant model, Takahashi et al.
found that the inner cells of tumor spher-
oids generated high levels of ROS, which
triggered the TRPA1-mediated Ca2+
influx required for inner cell survival.
Notably, TRPA1 inhibition did not alter
the ROS levels in inner spheroid cells.
Thus, in a mechanism distinct from a
ROS scavenging antioxidant defense
program, TRPA1 oxidization under high
levels of ROS allows for an increase in
Ca2+ into cells to promote cancer cell
survival.

Figure 1. ROS-Mediated TRPA1-Dependent Ca2+ Signaling Promotes Cancer Cell Survival
Cancer cells need to adapt to high levels of reactive oxygen species (ROS) in order to survive. The
transcription factor NRF2 induces expression of the TRPA1 channel. Hydrogen peroxide (H2O2), a type of
ROS produced by the mitochondria, can oxidize critical cysteine residues on TRPA1 to activate the
protein. As a result, intracellular calcium (Ca2+) concentrations increase, activating Ca2+-dependent anti-
apoptotic and pro-survival signaling cascades involving the MCL-1 protein. Thus, ROS can promote
cancer cell survival through Ca2+ signaling.
To better understand how TRPA1 sup-
ports cancer cell survival, Takahashi et al.
(2018) examined the role of TRPA1 in
anchorage-independent growth, a key
feature of anoikis resistance, cell survival
in the absence of extracellular matrix
(ECM) attachment, and tumor metas-
tasis. The authors used the antioxidant
N-Acetyl-L-cysteine (NAC) to show that
ECM detachment-mediated ROS pro-
duction was required for the increase in
intracellular Ca2+ by TRPA1. Further-
more, in a soft agar assay, inhibition of
TRPA1 reduced the colony number and
average colony size of HCC1569 breast
cancer cells, indicating the necessity of
TRPA1 for anchorage-independent
growth. These data suggest that TRPA1
promotes oxidative stress tolerance and
cancer cell survival in part by suppress-
ing anoikis. Takahashi et al. also found
that TRPA1-mediated Ca2+ influx pro-
moted cancer cell resistance to ROS-
producing therapies like carboplatin.
Consistent with their in vitro data, the au-
thors used immunocompromised mice
to show that genetic knockdown and
pharmacological inhibition of TRPA1 sup-
pressed tumor growth and chemoresist-
ance in vivo. Furthermore, by using
950 Cancer Cell 33, June 11, 2018
reverse-phase protein array, the authors
identified without bias that TRPA1
activates Ca2+-dependent pro-survival
and anti-apoptotic programs involving
MCL-1 to mediate oxidative stress toler-
ance in cancer cells. Specifically,
TRPA1 increased the phosphorylation of
RAS-ERK, PI3K/AKT, and mTOR pro-
survival signaling proteins and increased
the expression of the anti-apoptotic pro-
tein MCL-1 in a Ca2+-CaM/PYK2-depen-
dent manner. Moreover, Takahashi et al.
found that TRPA1 expression in cancer
cells was directly regulated by the
redox-sensitive antioxidant master regu-
lator NRF2. Thus, in response to H2O2,
NRF2 is stabilized and translocated to
the nucleus, where it induces the expres-
sion of both canonical (antioxidants) and
non-canonical (TRPA1) oxidative stress
defense proteins.
An intriguing finding of this study was
that high levels of ROS oxidized specific
cysteine residues of TRPA1, not to in-
crease cancer cell antioxidant capacity
but to promote cancer cell survival
through the upregulation of Ca2+-depen-
dent anti-apoptotic pathways (Figure 1).
Thus, ROS can function as signaling mol-
ecules to promote cancer cell survival
Cancer Cell
Previews
through TRPA1. Because TRPA1 was
overexpressed relative to corresponding
normal tissue in numerous cancer types
(Takahashi et al., 2018), targeting
TRPA1, as opposed to directly manipu-
lating ROS levels through antioxidant
inhibition, may be a more selective and
effective anti-cancer therapy. To date,
several TRPA1 inhibitors have been
developed as pain and respiratory thera-
pies; however, their pharmacokinetics
are poor, with a high IC50 and a short
half-life (Chen and Hackos, 2015). Thus,
future work should be done to improve
the efficacy and utility of TRPA1 inhibi-
tors as anti-cancer agents. Combination
therapy may increase the efficacy of in-
hibiting TRPA1, because Takahashi
et al. (2018) showed in vivo that TRPA1
inhibitor treatment in combination with
carboplatin reduced tumor growth more
than either drug treatment alone.
Furthermore, because TRPA1 was
shown to be activated by ROS in a
NRF2-dependent manner, this study
warrants the potential use of a TRPA1 in-
hibitor with selectivity for hypoxic tumor
cells, which have high levels of ROS, or
for tumor cells with a constitutively active
NRF2, like a subset of lung adenocarci-
noma cells. Additionally, it will be of in-
terest to determine whether other cal-
cium channels can be activated by
ROS to promote cancer cell survival.
Nevertheless, the findings from Takaha-
shi et al. reveal the existence of a novel,
ROS-controlled TRPA1-mediated cell
survival mechanism and open up an
exciting new avenue for the treatment
of cancer.
ACKNOWLEDGMENTS
This work was supported by the NIH
(5R35CA197532-02 to N.S.C.).
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Epigenetic ConFUSION: SS18-SSX Fusion
Rewires BAF Complex to Activate
Bivalent Genes in Synovial Sarcoma
Marc A. Morgan1 and Ali Shilatifard1,*
1Simpson Querrey Center for Epigenetics, Department of Biochemistry and Molecular Genetics, Northwestern University Feinberg School of
Medicine, 320 E. Superior St., Chicago, IL 60611, USA
*Correspondence: ash@northwestern.edu
https://doi.org/10.1016/j.ccell.2018.05.011
In this issue of Cancer Cell, McBride and colleagues report that the synovial sarcoma SS18-SSX fusion drives
BAF complex recruitment to bivalent domains repressed by PRC2 complex to orchestrate aberrant transcrip-
tional activation. Redistribution of BAF localization is a major driver of synovial sarcoma proliferation and pre-
sents a promising therapeutic target.
‘‘Uninvited guests are often most
welcome when they leave.’’
-Aesop
A group of friends (the BAF complex) has
gathered for a dinner party, but on this
particular evening, one member (SS18)
has brought along an uninvited and prob-
lematic guest (SSX). One friend (BAF47)
cannot abide this unwelcome new-
comer’s antics and leaves the party. The
interloper hijacks the conversation and
takes it to inappropriate areas (Bivalent
Polycomb repressed genes). This derails
the entire evening and leaves a lingering
malignant mood. This allegorical scene
describes molecular mechanisms for the
intruding SS18-SSX fusion oncoprotein
in synovial sarcoma, as revealed in this
issue of Cancer Cell (McBride et al.,
2018). Expelling the unwanted visitor,
SS18-SSX, restores normal BAF complex
function and reestablishes Polycomb
repression of bivalent genes. Importantly,
the authors reveal that eliminating SS18-
human prostate cancer stromal cells. Cancer
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ulation of oxidative stress as an anticancer strat-
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Paulsen, C.E., Armache, J.P., Gao, Y., Cheng, Y.,
and Julius, D. (2015). Structure of the TRPA1 ion
channel suggests regulatory mechanisms. Nature
520, 511–517.
SSX’s deviant gain-of-function behavior
is the primary requirement for halting
synovial sarcoma proliferation, rather
than restoring BAF47 (also known as
SMARCB1, SNF5, or INI1) to the BAF
complex. These findings shed new light
onto synovial sarcoma pathogenesis and
have important implications for the devel-
opment of targeted therapies.
Synovial sarcoma is a soft-tissue tumor
characterized by a reciprocal t(X;18)
translocation involving the SS18 gene
and one of three related SSX genes
(SSX1, SSX2, or SSX4) that occurs pre-
dominantly in young adults. SS18-SSX fu-
sions contain most of the SS18 protein
sequence followed by additional amino
acids derived from SSX. SS18 normally
participates as a component of the
multi-subunit BAF (mammalian SWI/
SNF) ATP-dependent chromatin-remod-
eling complex, thus implicating the dereg-
ulation of chromatin-based gene-regula-
tory systems as a central driver of
synovial sarcoma pathogenesis (Kadoch
and Crabtree, 2015). Despite SS18-
Cancer Cell 33, June 11, 2018 ª 2018 Published by Elsevier Inc. 951
Reczek, C.R., and Chandel, N.S. (2017). The two
faces of reactive oxygen species in cancer. Annu.
Rev. Cancer Biol. 1, 79–98.
Takahashi, N., Chen, H.Y., Harris, I.S., Stover,
D.G., Selfors, L.M., Bronson, R.T., Deraedt, T.,
Cichowski, K., Welm, A.L., Mori, Y., et al.
(2018). Cancer cells co-opt the neuronal redox-
sensing channel TRPA1 to promote oxidative
stress tolerance. Cancer Cell 33, this issue,
985–1003.
SSX’s status as a bona fide oncoprotein
(Haldar et al., 2007), until recently it re-
mained largely unknown how SS18-SSX
drives synovial sarcoma formation. In
2013, Kadoch and Crabtree published
their breakthrough discovery that SS18-
SSX alters the composition of BAF com-
plexes by evicting both wild-type SS18
and the tumor suppressor BAF47, thus
earmarking them for proteolytic degrada-
tion (Kadoch and Crabtree, 2013). These
biochemical alterations are accompanied
by the transcriptional activation of
oncogenic factors such as SOX2, which
are normally targeted for repression by
the Polycomb Repressive Complex 2
(PRC2). Importantly, removing SS18-
SSX restores wild-type SS18 and BAF47
to the BAF complex and is sufficient to
inhibit synovial sarcoma proliferation
(Kadoch and Crabtree, 2013), thus mak-
ing SS18-SSX an attractive therapeutic
target because of its involvement in both
tumor initiation and tumor maintenance.
In their current study, McBride and col-
leagues use patient-derived cell lines to