Abstract: A series of chemotherapeutic drugs that induce DNA damage, such as cisplatin (DDP), are standard clinical treatments for ovarian cancer, testicular cancer, and other diseases that lack effective targeted drug therapy. Drug resistance is one of the main factors limiting their application. Sensitizers can overcome the drug resistance of tumor cells, thereby enhancing the antitumor activity of chemotherapeutic drugs. In this study, we aimed to identify marketable drugs that could be potential chemotherapy sensitizers and explore the underlying mechanisms. We found that the alcohol withdrawal drug disulfiram (DSF) could significantly enhance the antitumor activity of DDP. JC-1 staining, propidium iodide (PI) staining, and western blotting confirmed that the combination of DSF and DDP could enhance the apoptosis of tumor cells. Subsequent RNA sequencing combined with Gene Set Enrichment Analysis (GSEA) pathway enrichment analysis and cell biology studies such as immunofluorescence suggested an underlying mechanism: DSF makes cells more vulnerable to DNA damage by inhibiting the Fanconi anemia (FA) repair pathway, exerting a sensitizing effect to DNA damaging agents including platinum chemotherapy drugs. Thus, our study illustrated the potential mechanism of action of DSF in enhancing the antitumor effect of DDP. This might provide an effective and safe solution for combating DDP resistance in clinical treatment.

双硫仑通过抑制范可尼贫血修复途径增加顺铂的抗肿瘤活性

[1]BaiZS, PengYL, YeXY, et al., 2022. Autophagy and cancer treatment: four functional forms of autophagy and their therapeutic applications. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(2):89-101.

[2]BrabecV, NovákováO, 2006. DNA binding mode of ruthenium complexes and relationship to tumor cell toxicity. Drug Resist Updat, 9(3):111-122.

[3]CastellaM, JacquemontC, ThompsonEL, et al., 2015. FANCI regulates recruitment of the FA core complex at sites of DNA damage independently of FANCD2. PLoS Genet, 11(10):e1005563.

[4]ChenSY, ChangYL, LiuST, et al., 2021. Differential cytotoxicity mechanisms of copper complexed with disulfiram in oral cancer cells. Int J Mol Sci, 22(7):3711.

[5]ChenZ, ChenJJ, 2021. Mass spectrometry-based protein‒protein interaction techniques and their applications in studies of DNA damage repair. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(1):1-20.

[6]FangL, QiH, WangP, et al., 2022. UPF1 increases amino acid levels and promotes cell proliferation in lung adenocarcinoma via the eIF2α-ATF4 axis. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(10):863-875.

[7]FengYL, LiuSC, ChenRD, et al., 2021. Target binding and residence: a new determinant of DNA double-strand break repair pathway choice in CRISPR/Cas9 genome editing. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(1):73-86.

[8]GalluzziL, SenovillaL, VitaleI, et al., 2012. Molecular mechanisms of cisplatin resistance. Oncogene, 31(15):1869-1883.

[9]GuoLM, CuiJ, WangHR, et al., 2021. Metformin enhances anti-cancer effects of cisplatin in meningioma through AMPK-mTOR signaling pathways. Mol Ther Oncolytics, 20:119-131.

[10]HalatschME, KastRE, Karpel-MasslerG, et al., 2021. A phase Ib/IIa trial of 9 repurposed drugs combined with temozolomide for the treatment of recurrent glioblastoma: CUSP9v3. Neuro-Oncol Adv, 3(1):vdab075.

[11]IshiaiM, 2021. Regulation of the fanconi anemia DNA repair pathway by phosphorylation and monoubiquitination. Genes, 12(11):1763.

[12]JangraA, ChoiSA, YangJ, et al., 2020. Disulfiram potentiates the anticancer effect of cisplatin in atypical teratoid/rhabdoid tumors (AT/RT). Cancer Lett, 486:38-45.

[13]KimSK, KimH, LeeDH, et al., 2013. Reversing the intract

[14]able nature of pancreatic cancer by selectively targeting ALDH-high, therapy-resistant cancer cells. PLoS ONE, 8(10):e78130.

[15]KitaY, HamadaA, SaitoR, et al., 2019. Systematic chemical screening identifies disulfiram as a repurposed drug that enhances sensitivity to cisplatin in bladder cancer: a summary of preclinical studies. Br J Cancer, 121(12):‍1027-1038.

[16]LeeJO, KangMJ, ByunWS, et al., 2019. Metformin overcomes resistance to cisplatin in triple-negative breast cancer (TNBC) cells by targeting RAD51. Breast Cancer Res, 21:115.

[17]LiangZR, ZhangT, ZhanT, et al., 2021. Metformin alleviates cisplatin-induced ototoxicity by autophagy induction possibly via the AMPK/FOXO3a pathway. J Neurophysiol, 125(4):1202-1212.

[18]LiuCC, WuCL, LinMX, et al., 2021. Disulfiram sensitizes a therapeutic-resistant glioblastoma to the TGF-β receptor inhibitor. Int J Mol Sci, 22(19):10496.

[19]MajeraD, SkrottZ, ChromaK, et al., 2020. Targeting the NPL4 adaptor of p97/VCP segregase by disulfiram as an emerging cancer vulnerability evokes replication stress and DNA damage while silencing the ATR pathway. Cells, 9(2):469.

[20]MakovecT, 2019. Cisplatin and beyond: molecular mechanisms of action and drug resistance development in cancer chemotherapy. Radiol Oncol, 53(2):148-158.

[21]MinKA, YuFQ, YangVC, et al., 2010. Transcellular transport of heparin-coated magnetic iron oxide nanoparticles (Hep-MION) under the influence of an applied magnetic field. Pharmaceutics, 2(2):119-135.

[22]NakanoT, WarnerKA, OklejasAE, et al., 2021. mTOR inhib

[23]ition ablates cisplatin-resistant salivary gland cancer stem cells. J Dent Res, 100(4):377-386.

[24]NechushtanH, HamamrehY, NidalS, et al., 2015. A phase IIb trial assessing the addition of disulfiram to chemotherapy for the treatment of metastatic non-small cell lung cancer. Oncologist, 20(4):366-367.

[25]ParkYM, GoYY, ShinSH, et al., 2018. Anti-cancer effects of disulfiram in head and neck squamous cell carcinoma via autophagic cell death. PLoS ONE, 13(9):e0203069.

[26]QiX, YanDH, ZuoJC, et al., 2021. Development of a novel chemokine signaling-based multigene signature to predict prognosis and therapeutic response in colorectal cancer. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(12):1053-1059.

[27]RenXY, LiYC, ZhouY, et al., 2021. Overcoming the compensatory elevation of NRF2 renders hepatocellular carcinoma cells more vulnerable to disulfiram/copper-induced ferroptosis. Redox Biol, 46:102122.

[28]RochaCRR, SilvaMM, QuinetA, et al., 2018. DNA repair pathways and cisplatin resistance: an intimate relationship. Clinics, 73(S1):e478s.

[29]SchmidtovaS, KalavskaK, GercakovaK, et al., 2019. Disulfiram overcomes cisplatin resistance in human embryonal carcinoma cells. Cancers, 11(9):1224.

[30]SkrottZ, MistrikM, AndersenKK, et al., 2017. Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4. Nature, 552(7684):194-199.

[31]SkrottZ, MajeraD, GurskyJ, et al., 2019. Disulfiram’s anti-cancer activity reflects targeting NPL4, not inhibition of aldehyde dehydrogenase. Oncogene, 38(40):6711-6722.

[32]SmogorzewskaA, MatsuokaS, VinciguerraP, et al., 2007. Identification of the FANCI protein, a monoubiquitinated FANCD2 paralog required for DNA repair. Cell, 129(2):289-301.

[33]WangK, MichelakosT, WangB, et al., 2021. Targeting cancer stem cells by disulfiram and copper sensitizes radioresistant chondrosarcoma to radiation. Cancer Lett, 505:37-48.

[34]WuQ, ZhangMY, WenYM, et al., 2022. Identifying chronic alcoholism drug disulfiram as a potent DJ-1 inhibitor for cancer therapeutics. Eur J Pharmacol, 926:175035.

[35]XiaL, LinHX, ZhouYM, et al., 2022. ZNF750 facilitates carcinogenesis via promoting the expression of long non-coding RNA CYTOR and influences pharmacotherapy response in colon adenocarcinoma. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(7):587-596.

[36]XuYQ, ZhouQ, FengXL, et al., 2020. Disulfiram/copper markedly induced myeloma cell apoptosis through activation of JNK and intrinsic and extrinsic apoptosis pathways. Biomed Pharmacother, 126:110048.

[37]YangYY, Lindsey-BoltzLA, VaughnCM, et al., 2021. Circadian clock, carcinogenesis, chronochemotherapy connections. J Biol Chem, 297(3):101068.

[38]YangZ, GuoF, AlbersAE, et al., 2019. Disulfiram modulates ROS accumulation and overcomes synergistically cisplatin resistance in breast cancer cell lines. Biomed Pharmacother, 113:108727.

[39]YardeDN, OliveiraV, MathewsL, et al., 2009. Targeting the Fanconi anemia/BRCA pathway circumvents drug resistance in multiple myeloma. Cancer Res, 69(24):9367-9375.

[40]ZhangHJ, ChenD, RinglerJ, et al., 2010. Disulfiram treatment facilitates phosphoinositide 3-kinase inhibition in human breast cancer cells in vitro and in vivo. Cancer Res, 70(10):3996-4004.

[41]ZhaoLJ, TengB, WenLJ, et al., 2014. mTOR inhibitor AZD8055 inhibits proliferation and induces apoptosis in laryngeal carcinoma. Int J Clin Exp Med, 7(2):337-347.

[42]ZhongWW, WangDJ, YaoB, et al., 2021. Integrative analysis of prognostic long non-coding RNAs with copy number variation in bladder cancer. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(8):664-681.

[43]ZirjacksL, StranskyN, KlumppL, et al., 2021. Repurposing disulfiram for targeting of glioblastoma stem cells: an in vitro study. Biomolecules, 11(11):1561.