Esophageal squamous cell carcinoma (ESCC) is among the most lethal malignancies worldwide, with a five-year survival rate below 20%. Ferroptosis—a regulated form of cell death driven by iron-dependent lipid peroxidation—has emerged as a promising therapeutic strategy, yet its regulation in ESCC remains poorly understood. We investigated the role of tripartite motif-containing 31 (TRIM31), an E3 ubiquitin ligase, in ESCC progression and ferroptosis.
Results:
TRIM31 expression was significantly elevated in ESCC tissues compared with normal esophageal tissues (The Cancer Genome Atlas and Genotype-Tissue Expression datasets; p < 0.001) and correlated with advanced Tumor, Node, and Metastasis (TNM) stage (p = 0.004), lymph node metastasis (p = 0.024), and poor overall survival (p = 0.0027). Functional assays revealed that TRIM31 knockdown reduced ESCC cell proliferation, impaired colony formation, and suppressed migration (p < 0.01). In vivo, TRIM31 silencing decreased xenograft tumor volume by over 70% and Ki-67 expression by 73%. Mechanistically, TRIM31 directly interacted with VDAC1 via its coiled-coil domain, promoting VDAC1 ubiquitination and proteasomal degradation. This interaction inhibited ferroptosis, as evidenced by increased lipid reactive oxygen species, elevated intracellular Fe2+ levels, and mitochondrial damage upon TRIM31 knockdown. Conversely, TRIM31 overexpression attenuated ferroptosis induced by RSL3. We further identified hepatocyte nuclear factor 4 alpha as a transcriptional activator of TRIM31, binding to its promoter region. Importantly, TRIM31 knockdown synergized with the ferroptosis inducer imidazole ketone erastin, achieving a 90% reduction in tumor growth without significant toxicity.
Conclusion:
TRIM31 promotes ESCC progression by degrading VDAC1 and suppressing ferroptosis. Targeting TRIM31 enhances ferroptosis-based therapy and represents a novel, clinically actionable strategy for ESCC treatment. Antioxid. Redox Signal. 44, 550–571.
ArifT, Shteinfer-KuzmineA, Shoshan-BarmatzV. Decoding cancer through silencing the mitochondrial gatekeeper VDAC1. Biomolecules, 2024; 14(10):1304.
3.
BrayF, LaversanneM, SungH, et al.Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2024; 74(3):229–263.
4.
CamaraA, ZhouY, WenPC, et al.Mitochondrial VDAC1: A key gatekeeper as potential therapeutic target. Front Physiol, 2017; 8:460.
5.
ChenX, KangR, KroemerG, et al.Broadening horizons: The role of ferroptosis in cancer. Nat Rev Clin Oncol, 2021; 18(5):280–296.
6.
ChenY, FangZM, YiX, et al.The interaction between ferroptosis and inflammatory signaling pathways. Cell Death Dis, 2023; 14(3):205.
7.
ChenZ, WangW, HouJ, et al.NEDD4L contributes to ferroptosis and cell growth inhibition in esophageal squamous cell carcinoma by facilitating xCT ubiquitination. Cell Death Discov, 2024; 10(1):473.
8.
ConradM, WahidaA. The many paths ascending to ferroptosis. Nat Chem Biol, 2025; 21(1):18–19.
9.
DengNH, TianZ, ZouYJ, et al.E3 ubiquitin ligase TRIM31: A potential therapeutic target. Biomed Pharmacother, 2024; 176:116846.
10.
FengQ, NieF, GanL, et al.Tripartite motif 31 drives gastric cancer cell proliferation and invasion through activating the Wnt/β-catenin pathway by regulating Axin1 protein stability. Sci Rep, 2023; 13(1):20099.
11.
GaoG, ZhangX. Broadening horizons: Research on ferroptosis in lung cancer and its potential therapeutic targets. Front Immunol, 2025; 16:1542844.
12.
GeL, SunM, XuD, et al.The E3 ubiquitin ligase TRIM31 enhances the progression of esophageal cancer through the ubiquitination of BBOX1. Cell Signal, 2025; 135:111975; doi: 10.1016/j.cellsig.2025.111975
13.
GuoP, MaX, ZhaoW, et al.TRIM31 is upregulated in hepatocellular carcinoma and promotes disease progression by inducing ubiquitination of TSC1-TSC2 complex. Oncogene, 2018; 37(4):478–488.
14.
GuoP, QiuY, MaX, et al.Tripartite motif 31 promotes resistance to anoikis of hepatocarcinoma cells through regulation of p53-AMPK axis. Exp Cell Res, 2018; 368(1):59–66; doi: 10.1016/j.yexcr.2018.04.013
15.
GuoY, LiQ, ZhaoG, et al.Loss of TRIM31 promotes breast cancer progression through regulating K48- and K63-linked ubiquitination of p53. Cell Death Dis, 2021; 12(10):945.
16.
GuoY, LinP, HuaY, et al.TRIM31: A molecule with a dual role in cancer. Front Oncol, 2022; 12:1047177.
17.
HuH, GuoL, OverholserJ, et al.Mitochondrial VDAC1: A potential therapeutic target of inflammation-related diseases and clinical opportunities. Cells, 2022; 11(19):3174.
18.
JiJ, ZhouC, WuJ, et al.Expression pattern of CDK12 protein in gastric cancer and its positive correlation with CD8(+) cell density and CCL12 expression. Int J Med Sci, 2019; 16(8):1142–1148.
19.
JiangK, YinX, ZhangQ, et al.STC2 activates PRMT5 to induce radioresistance through DNA damage repair and ferroptosis pathways in esophageal squamous cell carcinoma. Redox Biol, 2023; 60:102626.
20.
JiangL, KonN, LiT, et al.Ferroptosis as a p53-mediated activity during tumour suppression. Nature, 2015; 520(7545):57–62.
21.
JiangW, ZhangB, XuJ, et al.Current status and perspectives of esophageal cancer: A comprehensive review. Cancer Commun (Lond), 2025; 45(3):281–331.
22.
JiangX, StockwellBR, ConradM. Ferroptosis: Mechanisms, biology and role in disease. Nat Rev Mol Cell Biol, 2021; 22(4):266–282.
23.
JokhadzeN, DasA, DizonDS. Global cancer statistics: A healthy population relies on population health. CA Cancer J Clin, 2024; 74(3):224–226.
24.
LeiG, ZhangY, KoppulaP, et al.The role of ferroptosis in ionizing radiation-induced cell death and tumor suppression. Cell Res, 2020; 30(2):146–162.
25.
LeiG, ZhuangL, GanB. Targeting ferroptosis as a vulnerability in cancer. Nat Rev Cancer, 2022; 22(7):381–396.
26.
LemastersJJ. Evolution of voltage-dependent anion channel function: From molecular sieve to governator to actuator of ferroptosis. Front Oncol, 2017; 7:303.
27.
LiH, ZhangY, HaiJ, et al.Knockdown of TRIM31 suppresses proliferation and invasion of gallbladder cancer cells by down-regulating MMP2/9 through the PI3K/Akt signaling pathway. Biomed Pharmacother, 2018; 103:1272–1278; doi: 10.1016/j.biopha.2018.04.120
28.
LiY, LiZ, RanQ, et al.Sterols in ferroptosis: From molecular mechanisms to therapeutic strategies. Trends Mol Med, 2025; 31(1):36–49.
29.
LiuB, ZhangM, ChuH, et al.The ubiquitin E3 ligase TRIM31 promotes aggregation and activation of the signaling adaptor MAVS through Lys63-linked polyubiquitination. Nat Immunol, 2017; 18(2):214–224.
30.
LiuC, LiuZ, DongZ, et al.Multifaceted interplays between the essential players and lipid peroxidation in ferroptosis. J Genet Genomics, 2025; 52(9):1071–1081.
31.
LiuCC, LiHH, LinJH, et al.Esophageal cancer stem-like cells resist ferroptosis-induced cell death by active Hsp27-GPX4 pathway. Biomolecules, 2021; 12(1):48.
32.
LiuY, LuS, WuL, et al.The diversified role of mitochondria in ferroptosis in cancer. Cell Death Dis, 2023; 14(8):519.
33.
LuoX, GongHB, GaoHY, et al.Oxygenated phosphatidylethanolamine navigates phagocytosis of ferroptotic cells by interacting with TLR2. Cell Death Differ, 2021; 28(6):1971–1989.
34.
MaX, XiaoL, LiuL, et al.CD36-mediated ferroptosis dampens intratumoral CD8+ T cell effector function and impairs their antitumor ability. Cell Metab, 2021; 33(5):1001–1012.e5.
35.
NgNHJ, GhoshS, BokCM, et al.HNF4A and HNF1A exhibit tissue specific target gene regulation in pancreatic beta cells and hepatocytes. Nat Commun, 2024; 15(1):4288; doi: 10.1038/s41467-024-48647-w
36.
Nowicki-OsuchK, ZhuangL, JammulaS, et al.Molecular phenotyping reveals the identity of Barrett’s esophagus and its malignant transition. Science, 2021; 373(6556):760–767; doi: 10.1126/science.abd1449
37.
PanG, XiaY, HaoM, et al.EZH2 suppresses IR-induced ferroptosis by forming a co-repressor complex with HIF-1α to inhibit ACSL4: Targeting EZH2 enhances radiosensitivity in KDM6A-deficient esophageal squamous carcinoma. Cell Death Differ, 2025; 32(6):1026–1040.
38.
RaEA, LeeTA, Won KimS, et al.TRIM31 promotes Atg5/Atg7-independent autophagy in intestinal cells. Nat Commun, 2016; 7:11726.
39.
ReymondA, MeroniG, FantozziA, et al.The tripartite motif family identifies cell compartments. Embo J, 2001; 20(9):2140–2151.
40.
RyanSK, UgaldeCL, RollandAS, et al.Therapeutic inhibition of ferroptosis in neurodegenerative disease. Trends Pharmacol Sci, 2023; 44(10):674–688.
41.
SheikhM, RoshandelG, McCormackV, et al.Current status and future prospects for esophageal cancer. Cancers (Basel), 2023; 15(3):765.
42.
ShimadaH, NabeyaY, OkazumiS, et al.Prognostic significance of CYFRA 21-1 in patients with esophageal squamous cell carcinoma. J Am Coll Surg, 2003; 196(4):573–578.
43.
SongH, LiuB, HuaiW, et al.The E3 ubiquitin ligase TRIM31 attenuates NLRP3 inflammasome activation by promoting proteasomal degradation of NLRP3. Nat Commun, 2016; 7:13727.
44.
StraubeJ, LaneSW. The TRIM31 paradox: An unexpected benefit for leukemia stem cells. Haematologica, 2023; 108(8):1982–1983.
45.
TangD, ChenX, KangR, et al.Ferroptosis: Molecular mechanisms and health implications. Cell Res, 2021; 31(2):107–125.
46.
TortiSV, TortiFM. Iron and cancer: More ore to be mined. Nat Rev Cancer, 2013; 13(5):342–355.
47.
TunyasuvunakoolK, AdlerJ, WuZ, et al.Highly accurate protein structure prediction for the human proteome. Nature, 2021; 596(7873):590–596.
48.
VermaR, SattarRSA, KumarA, et al.Cross-talk between next generation sequencing methodologies to identify genomic signatures of esophageal cancer. Crit Rev Oncol Hematol, 2021; 162:103348.
49.
WangH, YaoL, GongY, et al.TRIM31 regulates chronic inflammation via NF-κB signal pathway to promote invasion and metastasis in colorectal cancer. Am J Transl Res, 2018; 10(4):1247–1259.
50.
WangY, WeiZ, PanK, et al.The function and mechanism of ferroptosis in cancer. Apoptosis, 2020; 25(11–12):786–798.
51.
WenQ, LiuJ, KangR, et al.The release and activity of HMGB1 in ferroptosis. Biochem Biophys Res Commun, 2019; 510(2):278–283.
52.
XiaW, LvY, ZouY, et al.The role of ferroptosis in colorectal cancer and its potential synergy with immunotherapy. Front Immunol, 2024; 15:1526749.
53.
YangF, XiaoY, DingJH, et al.Ferroptosis heterogeneity in triple-negative breast cancer reveals an innovative immunotherapy combination strategy. Cell Metab, 2023; 35(1):84–100.e8.
54.
YangJ, LuX, HaoJL, et al.VSTM2L protects prostate cancer cells against ferroptosis via inhibiting VDAC1 oligomerization and maintaining mitochondria homeostasis. Nat Commun, 2025; 16(1):1160.
55.
YangY, HuangX, ZhouL, et al.Clinical use of tumor biomarkers in prediction for prognosis and chemotherapeutic effect in esophageal squamous cell carcinoma. BMC Cancer, 2019; 19(1):526.
56.
YaoY, ChenZ, ZhangH, et al.Selenium-GPX4 axis protects follicular helper T cells from ferroptosis. Nat Immunol, 2021; 22(9):1127–1139.
57.
YuC, ChenS, GuoY, et al.Oncogenic TRIM31 confers gemcitabine resistance in pancreatic cancer via activating the NF-κB signaling pathway. Theranostics, 2018; 8(12):3224–3236.
58.
YuanS, WeiC, LiuG, et al.Sorafenib attenuates liver fibrosis by triggering hepatic stellate cell ferroptosis via HIF-1α/SLC7A11 pathway. Cell Prolif, 2022; 55(1):e13158.
59.
ZhangC, LiuX, JinS, et al.Ferroptosis in cancer therapy: A novel approach to reversing drug resistance. Mol Cancer, 2022; 21(1):47.
60.
ZhangJ, CaoL, WangX, et al.The E3 ubiquitin ligase TRIM31 plays a critical role in hypertensive nephropathy by promoting proteasomal degradation of MAP3K7 in the TGF-β1 signaling pathway. Cell Death Differ, 2022; 29(3):556–567.
61.
ZhangLL, ZhuWJ, ZhangXX, et al.Ferroptosis patterns and tumor microenvironment infiltration characterization in esophageal squamous cell cancer. Front Genet, 2022; 13:1047382.
62.
ZhangY, ShiJ, LiuX, et al.BAP1 links metabolic regulation of ferroptosis to tumour suppression. Nat Cell Biol, 2018; 20(10):1181–1192.
63.
ZhangY, TanH, DanielsJD, et al.Imidazole Ketone Erastin induces ferroptosis and slows tumor growth in a mouse lymphoma model. Cell Chem Biol, 2019; 26(5):623–633.e9.
64.
ZhaoL, ZhouX, XieF, et al.Ferroptosis in cancer and cancer immunotherapy. Cancer Commun (Lond), 2022; 42(2):88–116.
65.
ZhaoZ, SongX, WangY, et al.E3 ubiquitin ligase TRIM31 alleviates dopaminergic neurodegeneration by promoting proteasomal degradation of VDAC1 in Parkinson’s Disease model. Cell Death Differ, 2024; 31(11):1410–1421.
66.
ZhouQ, LiuT, QianW, et al.HNF4A-BAP31-VDAC1 axis synchronously regulates cell proliferation and ferroptosis in gastric cancer. Cell Death Dis, 2023; 14(6):356.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
