TMEM92 shields DDX3X from TTC3-mediated degradation to confer chemoresistance in triple-negative breast cancer

Abstract

Background: Triple-negative breast cancer (TNBC) remains a major clinical challenge because of its aggressive characteristics, limited targeted treatment options, and frequent chemoresistance. However, the molecular mechanisms governing protein stability that drive TNBC progression and therapeutic resistance remain incompletely understood.

Methods: TMEM92 expression and clinical relevance were evaluated using public datasets, patient specimens, and TNBC cell models. Loss-of-function, rescue, xenograft, protein interaction, and ubiquitination assays were performed to determine the biological function and molecular mechanism of TMEM92 in TNBC progression and cisplatin response.

Results: TMEM92 was prominently expressed in TNBC and correlated with poor prognosis. Functionally, depletion of TMEM92 suppressed TNBC cell proliferation, migration, invasion, and survival while promoting apoptosis in vitro and in vivo. Mechanistically, TMEM92 directly associated with DEAD-box helicase 3 X-linked (DDX3X) and protected it from degradation by the E3 ubiquitin ligase tetratricopeptide repeat domain 3 (TTC3). TMEM92 competitively prevented TTC3 binding to DDX3X, thereby inhibiting TTC3-mediated K48-linked ubiquitination and subsequent proteasomal degradation of DDX3X. Re-expression of DDX3X rescued the anti-tumor effects induced by TMEM92 knockdown. Therapeutically, TMEM92 targeting sensitized TNBC cells and xenograft tumors to cisplatin. TMEM92 knockout reduced the cisplatin IC₅₀ by 44.0% in MDA-MB-231 cells and 42.9% in BT-549 cells, and TMEM92 depletion enhanced cisplatin-induced tumor growth inhibition by approximately 70.6% compared with cisplatin alone.

Conclusions: This study identifies a novel TMEM92DDX3XTTC3 axis that regulates DDX3X protein stability and drives TNBC progression and chemoresistance, revealing a potential prognostic and therapeutic vulnerability in TNBC.

Keywords

cisplatin resistance / DDX3X / TMEM92 / triple-negative breast cancer / TTC3 / ubiquitination

References

[1] de Melo Gagliato D, Buzaid AC, Perez-Garcia J, Cortes J. Immunotherapy in breast cancer: current practice and clinical challenges. BioDrugs. 2020; 34(5): 611-623.

[2] Dogra AK, Prakash A, Gupta S, Gupta M. Prognostic significance and molecular classification of triple negative breast cancer: a systematic review. Eur J Breast Health. 2025; 21(2): 101-114.

[3] Ma T, Hao XM, Chen HD, et al. Predictive markers of rapid disease progression and chemotherapy resistance in triple-negative breast cancer patients following postoperative adjuvant therapy. Sci Rep. 2025; 15(1): 386.

[4] Zagami P, Carey LA. Triple negative breast cancer: pitfalls and progress. NPJ Breast Cancer. 2022; 8(1): 95.

[5] Ma J, Deng Y, Chen D, et al. Spatial immunophenotypes orchestrate prognosis in triple-negative breast cancer with Miller-Payne grade 4 following neoadjuvant chemotherapy. NPJ Breast Cancer. 2023; 9(1): 57.

[6] Yoder R, Kimler BF, Staley JM, et al. Impact of low versus negative estrogen/progesterone receptor status on clinico-pathologic characteristics and survival outcomes in HER2-negative breast cancer. NPJ Breast Cancer. 2022; 8(1): 80.

[7] Khadela A, Soni S, Megha K, et al. Contracting triple-negative breast cancer with immunotherapeutic armamentarium: recent advances and clinical prospects. Med Oncol. 2022; 40(1): 48.

[8] Obidiro O, Battogtokh G, Akala EO. Triple negative breast cancer treatment options and limitations: future outlook. Pharmaceutics. 2023; 15(7): 1796.

[9] Li Q, Ye Z, Wang G, et al. Natural products as novel therapeutic agents for triple-negative breast cancer: current evidence, mechanisms, challenges, and opportunities. Molecules. 2025; 30(6): 1201.

[10] Elmorsy EA, Saber S, Hamad RS, et al. Advances in understanding cisplatin-induced toxicity: molecular mechanisms and protective strategies. Eur J Pharm Sci. 2024; 203:106939.

[11] Pajewska M, Partyka O, Czerw A, et al. Advanced and metastatic triple negative breast cancer-potential new treatment. Cancers (Basel). 2025; 17(7): 1183.

[12] Eslami M, Memarsadeghi O, Davarpanah A, Arti A, Nayernia K, Behnam B. Overcoming chemotherapy resistance in metastatic cancer: a comprehensive review. Biomedicines. 2024; 12(1): 183.

[13] Khan SU, Fatima K, Aisha S, Malik F. Unveiling the mechanisms and challenges of cancer drug resistance. Cell Commun Signal. 2024; 22(1): 109.

[14] Kang H, Lee CJ. Transmembrane proteins with unknown function (TMEMs) as ion channels: electrophysiological properties, structure, and pathophysiological roles. Exp Mol Med. 2024; 56(4): 850-860.

[15] Zhang S, Wan X, Lv M, Li C, Chu Q, Wang G. TMEM92 acts as an immune-resistance and prognostic marker in pancreatic cancer from the perspective of predictive, preventive, and personalized medicine. Epma j. 2022; 13(3): 519-534.

[16] Girard N, Park K, Lee SH, et al. A brief report on stable disease among amivantamab-treated patients with post-platinum epidermal growth factor receptor exon 20 insertion-mutated non-small cell lung cancer: a response-based analysis from the CHRYSALIS study. Cancer Treat Res Commun. 2024; 40:100832.

[17] Zheng Y, Meng L, Qu L, et al. Co-targeting TMEM16A with a novel monoclonal antibody and EGFR with Cetuximab inhibits the growth and metastasis of esophageal squamous cell carcinoma. J Transl Med. 2024; 22(1): 1046.

[18] Chen C, Chen Z, Zhao J, et al. TMEM45A enhances palbociclib resistance and cellular glycolysis by activating AKT/mTOR signaling pathway in HR+ breast cancer. Cell Death Discov. 2025; 11(1): 47.

[19] Shi J, Zheng D, Yao B, Liu Q, Xu H, Piao H. Research progress on TMEM proteins in cancer progression and chemoresistance (Review). Int J Mol Med. 2025; 56(6): 219.

[20] Wu Z, Pan T, Li W, et al. Comprehensive pan-cancer analysis reveals prognostic implications of TMEM92 in the tumor immune microenvironment. Clin Transl Oncol. 2024; 26(10): 2701-2717.

[21] Herrera-Quiterio GA, Encarnación-Guevara S. The transmembrane proteins (TMEM) and their role in cell proliferation, migration, invasion, and epithelial-mesenchymal transition in cancer. Front Oncol. 2023; 13:1244740.

[22] Lin MZ, Teng LL, Sun XL, Zhang LP, Chen F, Yu LJ. Transmembrane protein 92 performs a tumor-promoting function in breast carcinoma by contributing to the cell growth, invasion, migration and epithelial-mesenchymal transition. Tissue Cell. 2020; 67:101415.

[23] Ye C, Ren S, Sadula A, et al. The expression characteristics of transmembrane protein genes in pancreatic ductal adenocarcinoma through comprehensive analysis of bulk and single-cell RNA sequence. Front Oncol. 2023; 13:1047377.

[24] Yao L, Hao Q, Wang M, et al. KLHL29-mediated DDX3X degradation promotes chemosensitivity by abrogating cell cycle checkpoint in triple-negative breast cancer. Oncogene. 2023; 42(47): 3514-3528.

[25] Zhang H, Mañán-Mejías PM, Miles HN, Putnam AA, MacGillivray LR, Ricke WA. DDX3X and stress granules: emerging players in cancer and drug resistance. Cancers (Basel). 2024; 16(6): 1131.

[26] Zeng Y, Tang X, Chen J, Kang X, Bai D. Optimizing total RNA extraction method for human and mice samples. PeerJ. 2024; 12:e18072.

[27] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001; 25(4): 402-408.

[28] Westerberg LJS, Dedic B, Näslund E, Thorell A, Spalding KL. Superior normalization using total protein for western blot analysis of human adipocytes. PLoS One. 2025; 20(7):e0328136.

[29] Juarez P, Martínez-Cerdeño V. Triple enzymatic immunochemistry for interneuron populations in postmortem human cerebral cortex. Heliyon. 2023; 9(10):e20626.

[30] Lei SY, Qin H, Liu S, Sheng J. Key nano-strategies for cisplatin resistance in advancing anti-cancer therapy. Int J Nanomedicine. 2025; 20: 13255-13292.

[31] Bi J, Wu Z, Zhang X, et al. TMEM25 inhibits monomeric EGFR-mediated STAT3 activation in basal state to suppress triple-negative breast cancer progression. Nat Commun. 2023; 14(1): 2342.

[32] Mirza L, Steventon L, Roylance R, et al. Regional differences in neo/adjuvant chemotherapy timing in patients with early-stage triple-negative breast cancer in England. Breast Cancer Res Treat. 2025; 209(1): 139-146.

[33] Xiong N, Wu H, Yu Z. Advancements and challenges in triple-negative breast cancer: a comprehensive review of therapeutic and diagnostic strategies. Front Oncol. 2024; 14:1405491.

[34] Wang Y, Qin Y, Wu C, et al. OSU-T315 overcomes immunosuppression in triple-negative breast cancer by targeting the ILK/NF-κB signaling pathway to enhance immunotherapeutic efficacy. Int Immunopharmacol. 2024; 143(Pt 3):113530.

[35] Joshi DC, Sharma A, Prasad S, et al. Novel therapeutic agents in clinical trials: emerging approaches in cancer therapy. Discov Oncol. 2024; 15(1): 342.

[36] Areewong S, Suppramote O, Prasopporn S, Jirawatnotai S. Exploiting acquired vulnerability to develop novel treatments for cholangiocarcinoma. Cancer Cell Int. 2024; 24(1): 362.

[37] Erler P, Kurcon T, Cho H, et al. Multi-armored allogeneic MUC1 CAR T cells enhance efficacy and safety in triple-negative breast cancer. Sci Adv. 2024; 10(35):eadn9857.

[38] Cui Y, Zhang W, Zeng X, Yang Y, Park SJ, Nakai K. Computational analysis of the functional impact of MHC-II-expressing triple-negative breast cancer. Front Immunol. 2024; 15:1497251.

[39] Shen L, Zhang J, Xu M, et al. DDX3 acts as a tumor suppressor in colorectal cancer as loss of DDX3 in advanced cancer promotes tumor progression by activating the MAPK pathway. Int J Biol Sci. 2022; 18(10): 3918-3933.

[40] Zhang W, Cao L, Yang J, et al. AEP-cleaved DDX3X induces alternative RNA splicing events to mediate cancer cell adaptation in harsh microenvironments. J Clin Invest. 2023; 134(3):e173299.

[41] Fan Z, Xu L, Gao Y, et al. The cytoplasmic-nuclear transport of DDX3X promotes immune-mediated liver injury in mice regulated by endoplasmic reticulum stress. Cell Death Dis. 2024; 15(9): 702.

[42] Khadivjam B, Bonneil É, Thibault P, Lippé R. RNA helicase DDX3X modulates herpes simplex virus 1 nuclear egress. Commun Biol. 2023; 6(1): 134.

[43] Leon-Ferre RA, Goetz MP. Advances in systemic therapies for triple negative breast cancer. BMJ. 2023; 381:e071674.

[44] Zhao Z, Li L, He M, Li Y, Ma X, Zhao B. Prognostic and predictive markers for early stage triple-negative breast cancer treated with platinum-based neoadjuvant chemotherapy. Cancer Med. 2024; 13(20):e70336.

[45] Marra A, Trapani D, Viale G, Criscitiello C, Curigliano G. Practical classification of triple-negative breast cancer: intratumoral heterogeneity, mechanisms of drug resistance, and novel therapies. NPJ Breast Cancer. 2020; 6: 54.

[46] Fu R, Zhao B, Chen M, et al. Moving beyond cisplatin resistance: mechanisms, challenges, and prospects for overcoming recurrence in clinical cancer therapy. Med Oncol. 2023; 41(1): 9.

[47] Pompella A, Corti A, Visvikis A. Redox mechanisms in cisplatin resistance of cancer cells: the twofold role of gamma-glutamyltransferase 1 (GGT1). Front Oncol. 2022; 12:920316.

[48] Wang G, Duan P, Wei Z, Liu F. Curcumin sensitizes carboplatin treatment in triple negative breast cancer through reactive oxygen species induced DNA repair pathway. Mol Biol Rep. 2022; 49(4): 3259-3270.

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2026 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.