{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T15:27:58Z","timestamp":1767713278280,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T00:00:00Z","timestamp":1762819200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation of Kore","award":["2022R1A2C2003513"],"award-info":[{"award-number":["2022R1A2C2003513"]}]},{"name":"Catholic Medical Center Research Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Tyrosine kinase inhibitors (TKIs), such as sunitinib and sorafenib, are standard treatments for renal cell carcinoma (RCC). However, most patients treated with these drugs eventually develop drug resistance and relapse; therefore, new treatment options for RCC are urgently required. Recent studies have focused on combination therapies targeting distinct molecular pathways that may produce synergistic effects and help overcome drug resistance in RCC. Niclosamide, an anthelmintic agent, is effective against various cancers; however, its potential in combination with sunitinib for treating RCC has not been evaluated. In this study, we assessed the therapeutic efficacy of niclosamide in combination with sunitinib against RCC and explored the underlying molecular mechanisms. Niclosamide alone inhibited RCC cell proliferation, whereas its combination with sunitinib produced a synergistic anticancer effect, both in vitro and in vivo. RNA sequencing (RNA-seq) and bioinformatic analyses showed that niclosamide modulated critical pathways, including BRIP1- and FANCA-mediated DNA repair and E2F2-regulated cell cycle progression. These findings provide proof-of-concept that niclosamide enhances TKI efficacy through modulation of DNA repair and cell cycle pathways, supporting the rationale for DNA damage response (DDR)-targeted combination strategies in RCC.<\/jats:p>","DOI":"10.3390\/ijms262210922","type":"journal-article","created":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T12:44:55Z","timestamp":1762865095000},"page":"10922","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Novel Therapeutic Strategy for Renal Cell Carcinoma: Niclosamide Enhances Sunitinib Efficacy via DNA Repair and Cell Cycle Pathways"],"prefix":"10.3390","volume":"26","author":[{"given":"Ae Ryang","family":"Jung","sequence":"first","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ga Eun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mee Young","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seung Ah","family":"Rhew","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8496-8825","authenticated-orcid":false,"given":"Dongho","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"U-Syn","family":"Ha","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sung-Hoo","family":"Hong","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6775-1157","authenticated-orcid":false,"given":"Ji Youl","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sae Woong","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4860-6127","authenticated-orcid":false,"given":"Yong Hyun","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Urology, Seoul St. Mary\u2019s Hospital, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.eururo.2018.08.036","article-title":"Epidemiology of Renal Cell Carcinoma","volume":"75","author":"Capitanio","year":"2019","journal-title":"Eur. Urol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"17009","DOI":"10.1038\/nrdp.2017.9","article-title":"Renal cell carcinoma","volume":"3","author":"Hsieh","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.semcancer.2012.06.001","article-title":"The VHL\/HIF axis in clear cell renal carcinoma","volume":"23","author":"Shen","year":"2013","journal-title":"Semin. Cancer Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1038\/nrd2380","article-title":"Molecular basis for sunitinib efficacy and future clinical development","volume":"6","author":"Faivre","year":"2007","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15586\/jkcvhl.2014.7","article-title":"Sunitinib resistance in renal cell carcinoma","volume":"1","author":"Morais","year":"2014","journal-title":"J. Kidney Cancer VHL"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"17588359211034708","DOI":"10.1177\/17588359211034708","article-title":"New approaches to first-line treatment of advanced renal cell carcinoma","volume":"13","author":"George","year":"2021","journal-title":"Ther. Adv. Med. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"100110","DOI":"10.1016\/j.crphar.2022.100110","article-title":"Evaluation of synergism in drug combinations and reference models for future orientations in oncology","volume":"3","author":"Duarte","year":"2022","journal-title":"Curr. Res. Pharmacol. Drug Discov."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/s41416-023-02502-9","article-title":"Unlocking hidden potential: Advancements, approaches, and obstacles in repurposing drugs for cancer therapy","volume":"130","author":"Weth","year":"2024","journal-title":"Br. J. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.canlet.2014.04.003","article-title":"Multi-targeted therapy of cancer by niclosamide: A new application for an old drug","volume":"349","author":"Li","year":"2014","journal-title":"Cancer Lett."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Xu, Q., Krause, M., Samoylenko, A., and Vainio, S. (2016). Wnt Signaling in Renal Cell Carcinoma. Cancers, 8.","DOI":"10.3390\/cancers8060057"},{"key":"ref_11","first-page":"2944","article-title":"Niclosamide exerts anticancer effects through inhibition of the FOXM1-mediated DNA damage response in prostate cancer","volume":"11","author":"Kim","year":"2021","journal-title":"Am. J. Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1159\/000490140","article-title":"Niclosamide Exhibits Potent Anticancer Activity and Synergizes with Sorafenib in Human Renal Cell Cancer Cells","volume":"47","author":"Yu","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1436","DOI":"10.1186\/s40064-016-3153-x","article-title":"Niclosamide suppresses renal cell carcinoma by inhibiting Wnt\/beta-catenin and inducing mitochondrial dysfunctions","volume":"5","author":"Zhao","year":"2016","journal-title":"Springerplus"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"W488","DOI":"10.1093\/nar\/gkaa216","article-title":"SynergyFinder 2.0: Visual analytics of multi-drug combination synergies","volume":"48","author":"Ianevski","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1038\/nrm.2016.58","article-title":"Regulation of DNA double-strand break repair by ubiquitin and ubiquitin-like modifiers","volume":"17","author":"Schwertman","year":"2016","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5858","DOI":"10.1074\/jbc.273.10.5858","article-title":"DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139","volume":"273","author":"Rogakou","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1038\/ncb884","article-title":"DNA damage-induced G-M checkpoint activation by histone H2AX and 53BP1","volume":"4","author":"Chen","year":"2002","journal-title":"Nat. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"909","DOI":"10.15252\/embj.201693860","article-title":"Interplay between Fanconi anemia and homologous recombination pathways in genome integrity","volume":"35","author":"Michl","year":"2016","journal-title":"Embo J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2272334","DOI":"10.1080\/15384047.2023.2272334","article-title":"Combination therapy of niclosamide with gemcitabine inhibited cell proliferation and apoptosis via Wnt\/\u03b2-catenin\/c-Myc signaling pathway by inducing \u03b2-catenin ubiquitination in pancreatic cancer","volume":"24","author":"Kang","year":"2023","journal-title":"Cancer Biol. Ther."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/j.biopha.2017.05.097","article-title":"Niclosamide induces apoptosis through mitochondrial intrinsic pathway and inhibits migration and invasion in human thyroid cancer in vitro","volume":"92","author":"Yu","year":"2017","journal-title":"Biomed. Pharmacother."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4150","DOI":"10.7150\/jca.26948","article-title":"Niclosamide Inhibits Cell Growth and Enhances Drug Sensitivity of Hepatocellular Carcinoma Cells via STAT3 Signaling Pathway","volume":"9","author":"Wang","year":"2018","journal-title":"J. Cancer"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1089\/ars.2012.4938","article-title":"New Insights into the Link Between DNA Damage and Apoptosis","volume":"19","author":"Cianfanelli","year":"2013","journal-title":"Antioxid. Redox Signal."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.clgc.2019.05.004","article-title":"DNA Damage Response Pathway Alteration in Locally Advanced Clear-Cell Renal-Cell Carcinoma Is Associated With a Poor Outcome","volume":"17","author":"Na","year":"2019","journal-title":"Clin. Genitourin. Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"593039","DOI":"10.3389\/fgene.2020.593039","article-title":"The Clinical Significance of DNA Damage Repair Signatures in Clear Cell Renal Cell Carcinoma","volume":"11","author":"Guo","year":"2020","journal-title":"Front. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"60","DOI":"10.3892\/or.2022.8271","article-title":"Apigenin inhibits renal cell carcinoma cell proliferation through G2\/M phase cell cycle arrest","volume":"47","author":"Bao","year":"2022","journal-title":"Oncol. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1350","DOI":"10.3892\/or.2014.2965","article-title":"Kaempferol induces cell cycle arrest and apoptosis in renal cell carcinoma through EGFR\/p38 signaling","volume":"31","author":"Song","year":"2014","journal-title":"Oncol. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1007\/s00280-019-03999-9","article-title":"6-Gingerol induces cell-cycle G1-phase arrest through AKT-GSK 3\u03b2-cyclin D1 pathway in renal-cell carcinoma","volume":"85","author":"Xu","year":"2020","journal-title":"Cancer Chemother. Pharmacol."},{"key":"ref_28","first-page":"7403","article-title":"Dauricine inhibits viability and induces cell cycle arrest and apoptosis via inhibiting the PI3K\/Akt signaling pathway in renal cell carcinoma cells","volume":"17","author":"Zhang","year":"2018","journal-title":"Mol. Med. Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1758835920907504","DOI":"10.1177\/1758835920907504","article-title":"Tyrosine kinase inhibitors and immunotherapy combinations in renal cell carcinoma","volume":"12","author":"Rassy","year":"2020","journal-title":"Ther. Adv. Med. Oncol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13578-016-0134-2","article-title":"Maintenance of genome stability by Fanconi anemia proteins","volume":"7","author":"Palovcak","year":"2017","journal-title":"Cell Biosci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2764","DOI":"10.1038\/s41388-023-02800-9","article-title":"FANCA deficiency promotes leukaemic progression by allowing the emergence of cells carrying oncogenic driver mutations","volume":"42","author":"Pawlikowska","year":"2023","journal-title":"Oncogene"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"10477","DOI":"10.1096\/fj.201802439RR","article-title":"Elevated FANCA expression determines a worse prognosis in chronic lymphocytic leukemia and interferes with p53 function","volume":"33","author":"Yanez","year":"2019","journal-title":"FASEB J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"289","DOI":"10.2741\/e767","article-title":"BRIP1, a potential candidate gene in development of non-BRCA1\/2 breast cancer","volume":"8","author":"Ouhtit","year":"2016","journal-title":"Front. Biosci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1530\/EC-17-0173","article-title":"BRIP1 overexpression is correlated with clinical features and survival outcome of luminal breast cancer subtypes","volume":"7","author":"Gupta","year":"2018","journal-title":"Endocr. Connect."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.3748\/wjg.v26.i11.1197","article-title":"Landscape of BRIP1 molecular lesions in gastrointestinal cancers from published genomic studies","volume":"26","author":"Voutsadakis","year":"2020","journal-title":"World J. Gastroenterol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2810","DOI":"10.1038\/sj.onc.1208612","article-title":"The E2F transcriptional network: Old acquaintances with new faces","volume":"24","author":"Dimova","year":"2005","journal-title":"Oncogene"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1007\/s12032-022-01715-x","article-title":"E2F2 enhances the chemoresistance of pancreatic cancer to gemcitabine by regulating the cell cycle and upregulating the expression of RRM2","volume":"39","author":"Liu","year":"2022","journal-title":"Med. Oncol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"600","DOI":"10.18632\/genesandcancer.143","article-title":"Clinical performance of E2Fs 1-3 in kidney clear cell renal cancer, evidence from bioinformatics analysis","volume":"8","author":"Liang","year":"2017","journal-title":"Genes Cancer"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6377","DOI":"10.1038\/s41598-021-85969-x","article-title":"Phase Ib trial of reformulated niclosamide with abiraterone\/prednisone in men with castration-resistant prostate cancer","volume":"11","author":"Parikh","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1158\/0008-5472.CAN-09-3722","article-title":"Sunitinib Acts Primarily on Tumor Endothelium rather than Tumor Cells to Inhibit the Growth of Renal Cell Carcinoma","volume":"70","author":"Huang","year":"2010","journal-title":"Cancer Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1061","DOI":"10.1158\/1535-7163.MCT-17-0925","article-title":"MicroRNA-203 Inhibits Long Noncoding RNA HOTAIR and Regulates Tumorigenesis through Epithelial-to-mesenchymal Transition Pathway in Renal Cell Carcinoma","volume":"17","author":"Dasgupta","year":"2018","journal-title":"Mol. Cancer Ther."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1093\/bioinformatics\/btp120","article-title":"TopHat: Discovering splice junctions with RNA-Seq","volume":"25","author":"Trapnell","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1542","DOI":"10.12688\/f1000research.8923.1","article-title":"TCGA Workflow: Analyze cancer genomics and epigenomics data using Bioconductor packages","volume":"5","author":"Silva","year":"2016","journal-title":"F1000Res"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/10922\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T12:45:30Z","timestamp":1762865130000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/10922"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,11]]},"references-count":43,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262210922"],"URL":"https:\/\/doi.org\/10.3390\/ijms262210922","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,11]]}}}