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Inhibition of IRE1α kinase increases ferroptosis resistance in triple‑negative breast cancer cells

  • Authors:
    • Shuo Xu
    • Si-Qi Yang
  • View Affiliations / Copyright

    Affiliations: School of Medicine and Health, Urban Vocational College of Sichuan, Chengdu, Sichuan 610110, P.R. China, State Key Laboratory of Biotherapy, Sichuan University, Chengdu, Sichuan 610041, P.R. China
    Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 130
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    Published online on: February 10, 2026
       https://doi.org/10.3892/ol.2026.15483
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Abstract

Ferroptosis is a regulated form of cell death that serves a pivotal role in tumor suppression. Whilst the ribonuclease activity of inositol‑requiring enzyme 1α (IRE1α) is associated with the regulation of ferroptosis, the potential involvement of its kinase domain in this process remains elusive. Thus, the present study aimed to investigate the specific role of the IRE1α kinase domain in regulating ferroptosis in breast cancer, particularly in the triple‑negative breast cancer (TNBC) subtype. To this end, it employed a combination of bioinformatic analysis of clinical datasets, pharmacological inhibition of IRE1α kinase and genetic overexpression models in TNBC cell lines. The present study demonstrated that endoplasmic reticulum to nucleus signaling 1 (ERN1; the gene encoding IRE1α) was significantly downregulated in breast cancer compared with that in normal tissues, and that lower ERN1 levels were associated with a worse prognosis of patients with breast cancer. This association persisted in human epidermal growth factor receptor 2‑positive and TNBC subtypes. In TNBC, IRE1α kinase inhibitors (APY29 and sunitinib) markedly inhibited ferroptosis induced by system Xc‑ inhibition. Moreover, by constructing overexpression models of wild‑type IRE1α (IRE1α‑WT) and a kinase‑dead mutant (IRE1α‑K599A), it was demonstrated that IRE1α‑WT overexpression significantly enhanced sensitivity to ferroptosis, whereas the kinase‑dead mutant had no significant effect. Mechanistically, IRE1α kinase inhibition upregulated solute carrier family 7 member 11 (also known as xCT) expression and promoted glutathione (GSH) synthesis, thereby suppressing ferroptosis. Collectively, the present study reveals a new function of IRE1α kinase in the regulation of ferroptosis, highlighting the critical regulatory role of the IRE1α kinase‑xCT‑GSH axis in ferroptosis in TNBC. Thus, IRE1α kinase may have potential as a therapeutic target.
View Figures

Figure 1

IRE1α is associated with BRCA
progression. (A) UALCAN analysis of the level of ERN1 in
BRCA. (B) UALCAN analysis of the level of ERN1 in BRCA based
on BRCA subclasses. The Kaplan-Meier Plotter was used to analyze
the outcomes patients with BRCA, and the differences in
recurrence-free survival were compared between groups stratified by
ERN1 status: (C) Overall BRCA, (D) HER2-positive breast
cancer subset and (E) triple negative BRCA subset. IRE1α,
inositol-requiring enzyme 1α; UALCAN, University of ALabama at
Birmingham CANcer data analysis Portal; ERN1, endoplasmic
reticulum to nucleus signaling 1; BRCA, breast cancer; TCGA, The
Cancer Genome Atlas; HER2, human epidermal growth factor receptor
2; HR, hazard ratio; ns, not significant.

Figure 2

APY29 antagonizes ferroptosis induced
by system Xc− inhibition. Cell viability in MDA-MB-231
cells treated with (A) erastin, (B) SAS, (C) glutamate or (D)
cystine restriction for 24 h in the presence or absence of APY29
(0.2 µM). Cell viability in MDA-MB-468 cells treated with (E)
erastin or (F) SAS for 24 h in the presence or absence of APY29
(0.2 µM). Cell viability in 4T1 cells treated with (G) erastin or
(H) SAS for 24 h in the presence or absence of APY29 (0.2 µM). Data
are presented as mean ± standard error of the mean (n=6).
****P<0.0001 vs. Control. SAS, sulfasalazine.

Figure 3

Sunitinib and KIRA6 antagonize
ferroptosis induced by system Xc− inhibition. Cell
viability in MDA-MB-231 cells treated with (A) erastin, (B) SAS,
(C) glutamate or (D) cystine restriction for 24 h in the presence
or absence of sunitinib (0.2 µM). Cell viability in MDA-MB-231
cells treated with (E) erastin, (F) SAS, (G) glutamate or (H)
cystine restriction for 24 h in the presence or absence of KIRA6
(0.5 µM). Data are presented as mean ± standard error of the mean
(n=6). ****P<0.0001 vs. Control. SAS, sulfasalazine; KIRA6,
inositol-requiring enzyme 1α kinase-inhibiting RNase
attenuator.

Figure 4

Overexpression of IRE1α-WT promotes
ferroptosis. Dose-response of (A) erastin-, (B) SAS-, (C)
glutamate- or (D) cystine restriction-induced cytotoxicity in
MDA-MB-231 cells overexpressing empty vector, WT IRE1α or
kinase-dead IRE1α mutant. Data are presented as mean ± standard
error of the mean (n=6). ****P<0.0001 vs. IRE1α-WT+ vs. NC.
IRE1α, inositol-requiring enzyme 1α; WT, wildtype; NC, negative
control. SAS, sulfasalazine.

Figure 5

Pharmacological inhibition of IRE1α
kinase attenuates lipid peroxidation. MDA levels were measured in
MDA-MB-231 cells following 12-h treatment with (A) erastin (5 µM),
(B) SAS (1 mM), (C) glutamate (20 mM) or (D) cystine-restricted
medium, in the presence or absence of APY29 (0.2 µM). Lipid ROS
levels were measured in MDA-MB-231 cells following 6-h treatment
with (E) erastin (5 µM), (F) SAS (1 mM), (G) glutamate (20 mM) or
(H) cystine-restricted medium, in the presence or absence of APY29
(0.2 µM), with corresponding statistical histograms. Data are
presented as mean ± standard error of the mean (n=3). IRE1α,
inositol-requiring enzyme 1α; MDA, malondialdehyde; ROS, reactive
oxygen species; SAS, sulfasalazine.

Figure 6

Upregulation of IRE1α kinase
increases lipid peroxidation. MDA levels in MDA-MB-231 cells
overexpressing empty vector, wild-type IRE1α or kinase-dead IRE1α
mutant following 12-h treatment with (A) erastin (5 µM), (B) SAS (1
mM), (C) glutamate (20 mM) or (D) cystine-restricted medium. Lipid
ROS levels in MDA-MB-231 cells overexpressing empty vector,
wild-type IRE1α or kinase-dead IRE1α mutant following 6-h treatment
with (E) erastin (5 µM), (F) SAS (1 mM), (G) glutamate (20 mM) or
(H) cystine-restricted medium, with corresponding statistical
histograms. Data are presented as mean ± standard error of the mean
(n=3). IRE1α, inositol-requiring enzyme 1α; MDA, malondialdehyde;
ROS, reactive oxygen species; NC, negative control; WT, wildtype;
SAS, sulfasalazine.

Figure 7

IRE1α regulates GSH and participates
in ferroptosis. MDA-MB-231 cells were treated with APY29 (0.2 µM)
for 24 h, and the protein expression of (A) xCT, (B) ACSL4 and (C)
GPX4 was analyzed using western blotting. GSH levels were measured
in MDA-MB-231 cells following 12-h treatment with (D) erastin (5
µM), (E) SAS (1 mM), (F) glutamate (20 mM) or (G)
cystine-restricted medium, in the presence or absence of APY29 (0.2
µM). Cellular GSH levels were measured in MDA-MB-231 cells
overexpressing empty vector, wild-type IRE1α or the kinase-dead
IRE1α mutant following 12-h treatment with (H) erastin (5 µM), (I)
SAS (1 mM), (J) glutamate (20 mM) or (K) cystine-restricted medium.
Data are presented as mean ± standard error of the mean (n=3).
IRE1α, inositol-requiring enzyme 1α; GSH, glutathione; xCT, solute
carrier family 7 member 11; ACSL4, acyl-CoA synthetase long chain
family member 4; GPX4, glutathione peroxidase 4; NC, negative
control; WT, wildtype; ns, not significant; SAS, sulfasalazine.
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Copy and paste a formatted citation
Spandidos Publications style
Xu S and Yang S: Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells. Oncol Lett 31: 130, 2026.
APA
Xu, S., & Yang, S. (2026). Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells. Oncology Letters, 31, 130. https://doi.org/10.3892/ol.2026.15483
MLA
Xu, S., Yang, S."Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells". Oncology Letters 31.4 (2026): 130.
Chicago
Xu, S., Yang, S."Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells". Oncology Letters 31, no. 4 (2026): 130. https://doi.org/10.3892/ol.2026.15483
Copy and paste a formatted citation
x
Spandidos Publications style
Xu S and Yang S: Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells. Oncol Lett 31: 130, 2026.
APA
Xu, S., & Yang, S. (2026). Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells. Oncology Letters, 31, 130. https://doi.org/10.3892/ol.2026.15483
MLA
Xu, S., Yang, S."Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells". Oncology Letters 31.4 (2026): 130.
Chicago
Xu, S., Yang, S."Inhibition of IRE1&alpha; kinase increases ferroptosis resistance in triple‑negative breast cancer cells". Oncology Letters 31, no. 4 (2026): 130. https://doi.org/10.3892/ol.2026.15483
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