{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T11:16:41Z","timestamp":1781867801802,"version":"3.54.5"},"reference-count":50,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2023,12,18]],"date-time":"2023-12-18T00:00:00Z","timestamp":1702857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012283","name":"Chiayi Chang Gung Memorial Hospital","doi-asserted-by":"publisher","award":["CMRPG6H0381-83"],"award-info":[{"award-number":["CMRPG6H0381-83"]}],"id":[{"id":"10.13039\/501100012283","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bladder cancer is becoming one of the most common malignancies across the world. Although treatment strategy has been continuously improved, which has led to cisplatin-based chemotherapy becoming the standard medication, cancer recurrence and metastasis still occur in a high proportion of patients because of drug resistance. The high efficacy of regorafenib, a broad-spectrum kinase inhibitor, has been evidenced in treating a variety of advanced cancers. Hence, this study investigated whether regorafenib could also effectively antagonize the survival of cisplatin-resistant bladder cancer and elucidate the underlying mechanism. Two types of cisplatin-resistant bladder cancer cells, T24R1 and T24R2, were isolated from T24 cisplatin-sensitive bladder cancer cells. These cells were characterized, and T24R1- and T24R2-xenografted tumor mice were created to examine the therapeutic efficacy of regorafenib. T24R1 and T24R2 cells exhibited higher expression levels of epithelial\u2013mesenchymal transition (EMT) and stemness markers compared to the T24 cells, and regorafenib could simultaneously inhibit the viability and the expression of EMT\/stemness markers of both T24R1 and T24R2 cells. Moreover, regorafenib could efficiently arrest the cell cycle, promote apoptosis, and block the transmigration\/migration capabilities of both types of cells. Finally, regorafenib could significantly antagonize the growth of T24R1- and T24R2-xenografted tumors in mice. These results demonstrated the therapeutic efficacy of regorafenib in cisplatin-resistant bladder cancers. This study, thus, provides more insights into the mechanism of action of regorafenib and demonstrates its great potential in the future treatment of cisplatin-resistant advanced bladder cancer patients.<\/jats:p>","DOI":"10.3390\/ijms242417610","type":"journal-article","created":{"date-parts":[[2023,12,18]],"date-time":"2023-12-18T08:55:29Z","timestamp":1702889729000},"page":"17610","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Therapeutic Potential of Regorafenib in Cisplatin-Resistant Bladder Cancer with High Epithelial\u2013Mesenchymal Transition and Stemness Properties"],"prefix":"10.3390","volume":"24","author":[{"given":"Feng-Che","family":"Kuan","sequence":"first","affiliation":[{"name":"Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan"},{"name":"Division of Hematology and Oncology, Department of Medicine, Chang Gung Memorial Hospital Chiayi Branch, Chiayi 61363, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jhy-Ming","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Animal Science, National Chiayi University, Chiayi 60004, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5683-8406","authenticated-orcid":false,"given":"Yun-Ching","family":"Huang","sequence":"additional","affiliation":[{"name":"Division of Urology, Department of Surgery, Chang Gung Memorial Hospital Chiayi Branch, Chiayi 61363, Taiwan"},{"name":"Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2276-7785","authenticated-orcid":false,"given":"Shun-Fu","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Medical Research and Development, Chang Gung Memorial Hospital Chiayi Branch, Chiayi 61363, Taiwan"},{"name":"Center for General Education, Chiayi Chang Gung University of Science and Technology, Chiayi 61363, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5548-7675","authenticated-orcid":false,"given":"Chung-Sheng","family":"Shi","sequence":"additional","affiliation":[{"name":"Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan"},{"name":"Division of Colon and Rectal Surgery, Department of Surgery, Chang Gung Memorial Hospital Chiayi Branch, Chiayi 61363, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21708","article-title":"Cancer statistics, 2022","volume":"72","author":"Siegel","year":"2022","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Garc\u00eda-Caballero, M., Torres-Vargas, J.A., Marrero, A.D., Mart\u00ednez-Poveda, B., Medina, M., and Quesada, A.R. (2022). Angioprevention of Urologic Cancers by Plant-Derived Foods. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14020256"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1038\/s41598-021-04035-8","article-title":"In vitro assessment of intra-operative and post-operative environment in reducing bladder cancer recurrence","volume":"12","author":"Tse","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e31635","DOI":"10.1097\/MD.0000000000031635","article-title":"Metformin and bladder cancer: Drug repurposing as a potential tool for novel therapy: A review","volume":"101","author":"Feng","year":"2022","journal-title":"Medicine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1200\/JCO.2001.19.3.666","article-title":"Radical cystectomy in the treatment of invasive bladder cancer: Long-term results in 1054 patients","volume":"19","author":"Stein","year":"2001","journal-title":"J. Clin. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1634\/theoncologist.2015-0440","article-title":"Neoadjuvant chemotherapy for muscle-invasive bladder cancer: A systematic review and two-step meta-analysis","volume":"21","author":"Yin","year":"2016","journal-title":"Oncologist"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"63","DOI":"10.4111\/icu.2016.57.1.63","article-title":"Upregulated expression of BCL2, MCM7, and CCNE1 indicate cisplatin-resistance in the set of two human bladder cancer cell lines: T24 cisplatin sensitive and T24R2 cisplatin resistant bladder cancer cell lines","volume":"57","author":"Kim","year":"2016","journal-title":"Investig. Clin. Urol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"303","DOI":"10.2147\/JEP.S267383","article-title":"Advances in our understanding of the molecular mechanisms of action of csplatin in cancer therapy","volume":"13","author":"Tchounwou","year":"2021","journal-title":"J. Exp. Pharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"202","DOI":"10.4111\/icu.20230006","article-title":"Perioperative systemic therapy in muscle invasive bladder cancer: Current standard method, biomarkers and emerging strategies","volume":"64","author":"Kim","year":"2023","journal-title":"Investig. Clin. Urol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3152","DOI":"10.1080\/10667857.2022.2133073","article-title":"Gold nanoparticles loaded TNF-\u03b1 and CALNN peptide as a drug delivery system and promising therapeutic agent for breast cancer cells","volume":"37","author":"Jabir","year":"2022","journal-title":"Mater. Technol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Ibrahim, A.A., Kareem, M.M., Al-Noor, T.H., Al-Muhimeed, T., AlObaid, A.A., Albukhaty, S., Sulaiman, G.M., Jabir, M., Taqi, Z.J., and Sahib, U.I. (2021). Pt(II)-thiocarbohydrazone complex as cytotoxic agent and apoptosis inducer in caov-3 and HT-29 cells through the p53 and caspase-8 pathways. Pharmaceuticals, 14.","DOI":"10.3390\/ph14060509"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2115","DOI":"10.7150\/thno.69424","article-title":"Platinum-based drugs for cancer therapy and anti-tumor strategies","volume":"12","author":"Zhang","year":"2022","journal-title":"Theranostics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1038\/s41568-019-0213-x","article-title":"Controversies around epithelial-mesenchymal plasticity in cancer metastasis","volume":"19","author":"Williams","year":"2019","journal-title":"Nat. Rev. Cancer"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"25","DOI":"10.21037\/sci.2019.08.08","article-title":"Epithelial-mesenchymal plasticity-engaging stemness in an interplay of phenotypes","volume":"6","author":"Chin","year":"2019","journal-title":"Stem Cell Investig."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1186\/s13000-015-0263-7","article-title":"Gene expression of OCT4, SOX2, KLF4 and MYC (OSKM) induced pluripotent stem cells: Identification for potential mechanisms","volume":"10","author":"Cai","year":"2015","journal-title":"Diagn. Pathol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Tan, J., Wang, Y., Sun, L., Xu, S., Li, C., and Jin, X. (2022). The origin and evolution of bladder cancer stem cells. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.950241"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1038\/labinvest.2017.17","article-title":"Epithelial-mesenchymal transition promotes SOX2 and NANOG expression in bladder cancer","volume":"97","author":"Migita","year":"2017","journal-title":"Lab. Invest."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"260","DOI":"10.4252\/wjsc.v8.i8.260","article-title":"Epithelial plasticity in urothelial carcinoma: Current advancements and future challenges","volume":"8","author":"Garg","year":"2016","journal-title":"World J. Stem Cells"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"66668","DOI":"10.18632\/oncotarget.19112","article-title":"Bladder cancer stem cells: Clonal origin and therapeutic perspectives","volume":"8","author":"Li","year":"2017","journal-title":"Oncotarget"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1020675","DOI":"10.3389\/fonc.2022.1020675","article-title":"Impact of SOX2 function and regulation on therapy resistance in bladder cancer","volume":"12","author":"Chen","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1038\/s41573-021-00195-4","article-title":"Kinase drug discovery 20 years after imatinib: Progress and future directions","volume":"20","author":"Cohen","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1186\/s12943-018-0804-2","article-title":"Kinase-targeted cancer therapies: Progress, challenges and future directions","volume":"17","author":"Bhullar","year":"2018","journal-title":"Mol. Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"101993","DOI":"10.1016\/j.ctrv.2020.101993","article-title":"Evolving role of regorafenib for the treatment of advanced cancers","volume":"86","author":"Grothey","year":"2020","journal-title":"Cancer Treat. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5523","DOI":"10.2147\/CMAR.S305939","article-title":"Effect of regorafenib in delaying definitive deterioration in health-related quality of life in patients with advanced cancer of three different tumor types","volume":"13","author":"Hofheinz","year":"2021","journal-title":"Cancer Manag. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1002\/cam4.4430","article-title":"Regorafenib for Taiwanese patients with unresectable hepatocellular carcinoma after sorafenib failure: Impact of alpha-fetoprotein levels","volume":"11","author":"Hsu","year":"2022","journal-title":"Cancer Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.ejca.2021.06.027","article-title":"An open-label single-arm phase II study of regorafenib for the treatment of angiosarcoma","volume":"154","author":"Agulnik","year":"2021","journal-title":"Eur. J. Cancer"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3464","DOI":"10.1002\/cncr.32964","article-title":"A multi-institutional phase 2 trial of regorafenib in refractory advanced biliary tract cancer","volume":"126","author":"Kim","year":"2020","journal-title":"Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4009","DOI":"10.2217\/fon-2019-0480","article-title":"Regorafenib in patients with refractory metastatic pancreatic cancer: A Phase II study (RESOUND)","volume":"15","author":"Bozzarelli","year":"2019","journal-title":"Future Oncol."},{"key":"ref_29","first-page":"4919","article-title":"Regorafenib induces apoptosis and inhibits metastatic potential of human bladder carcinoma cells","volume":"37","author":"Hsu","year":"2017","journal-title":"Anticancer. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1002\/tox.22734","article-title":"Regorefenib induces extrinsic\/intrinsic apoptosis and inhibits MAPK\/NF-\u03baB-modulated tumor progression in bladder cancer in vitro and in vivo","volume":"34","author":"Chiang","year":"2019","journal-title":"Environ. Toxicol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1186\/s12964-020-00625-7","article-title":"Role of tyrosine kinases in bladder cancer progression: An overview","volume":"18","author":"Zangouei","year":"2020","journal-title":"Cell Commun. Signal"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"365","DOI":"10.3892\/mco.2017.1145","article-title":"Response to regorafenib at an initial dose of 120 mg as salvage therapy for metastatic colorectal cancer","volume":"6","author":"Osawa","year":"2017","journal-title":"Mol. Clin. Oncol."},{"key":"ref_33","first-page":"371","article-title":"Reduced dose of salvage-line regorafenib monotherapy for metastatic colorectal cancer in Japan","volume":"35","author":"Hirano","year":"2015","journal-title":"Anticancer. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1200\/JCO.19.03296","article-title":"Regorafenib plus nivolumab in patients with advanced gastric or colorectal cancer: An open-label, dose-escalation, and dose-expansion phase Ib trial (REGONIVO, EPOC1603)","volume":"38","author":"Fukuoka","year":"2020","journal-title":"J. Clin. Oncol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s12276-019-0308-1","article-title":"Anti-tumoral activity of single and combined regorafenib treatments in preclinical models of liver and gastrointestinal cancers","volume":"51","author":"Fondevila","year":"2019","journal-title":"Exp. Mol. Med."},{"key":"ref_36","first-page":"1241","article-title":"[Regorafenib versus S-1 plus bevacizumab for metastatic colorectal cancer as salvage line-A phase II study (OGSG1301)]","volume":"48","author":"Kodama","year":"2021","journal-title":"Gan To Kagaku Ryoho"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"912699","DOI":"10.3389\/fonc.2022.912699","article-title":"Neoadjuvant treatment in muscle-invasive bladder cancer: From the beginning to the latest developments","volume":"12","author":"Roviello","year":"2022","journal-title":"Front. Oncol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e2015808118","DOI":"10.1073\/pnas.2015808118","article-title":"ACTL6A promotes repair of cisplatin-induced DNA damage, a new mechanism of platinum resistance in cancer","volume":"118","author":"Xiao","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Kiss, R.C., Xia, F., and Acklin, S. (2021). Targeting DNA damage response and repair to enhance therapeutic index in cisplatin-based cancer treatment. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22158199"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"49447","DOI":"10.1007\/s11356-021-15759-5","article-title":"Role of multidrug resistance-associated proteins in cancer therapeutics: Past, present, and future perspectives","volume":"28","author":"Elfadadny","year":"2021","journal-title":"Environ. Sci. Pollut. Res. Int."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1002\/dvdy.24620","article-title":"Epithelial-mesenchymal transition and plasticity in the developmental basis of cancer and fibrosis","volume":"247","author":"Runyan","year":"2018","journal-title":"Dev. Dyn."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Dzobo, K., Senthebane, D.A., Ganz, C., Thomford, N.E., Wonkam, A., and Dandara, C. (2020). Advances in therapeutic targeting of cancer stem cells within the tumor microenvironment: An updated review. Cells, 9.","DOI":"10.20944\/preprints202005.0315.v1"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Abugomaa, A., Elbadawy, M., Yamawaki, H., Usui, T., and Sasaki, K. (2020). Emerging roles of cancer stem cells in bladder cancer progression, tumorigenesis, and resistance to chemotherapy: A potential therapeutic target for bladder cancer. Cells, 9.","DOI":"10.3390\/cells9010235"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"253","DOI":"10.2220\/biomedres.36.253","article-title":"Protein expression profile related to cisplatin resistance in bladder cancer cell lines detected by two-dimensional gel electrophoresis","volume":"36","author":"Taoka","year":"2015","journal-title":"Biomed. Res."},{"key":"ref_45","first-page":"2597","article-title":"Modified annexin V\/propidium iodide apoptosis assay for accurate assessment of cell death","volume":"50","author":"Rieger","year":"2011","journal-title":"J. Vis. Exp."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Wu, M.H., Hsu, W.B., Chen, M.H., and Shi, C.S. (2022). Inhibition of neddylation suppresses osteoclast differentiation and function in vitro and alleviates osteoporosis in vivo. Biomedicines, 10.","DOI":"10.3390\/biomedicines10102355"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1016\/j.ccell.2018.03.006","article-title":"lncRNA epigenetic landscape analysis identifies EPIC1 as an oncogenic incRNA that interacts with MYC and promotes cell-cycle progression in cancer","volume":"33","author":"Wang","year":"2018","journal-title":"Cancer Cell"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"5391","DOI":"10.1158\/1078-0432.CCR-14-2680","article-title":"Activation of EZH2 and SUZ12 regulated by E2F1 predicts the disease progression and aggressive characteristics of bladder cancer","volume":"21","author":"Lee","year":"2015","journal-title":"Clin. Cancer Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"736","DOI":"10.3892\/ijo.2017.3844","article-title":"Insulin induction instigates cell proliferation and metastasis in human colorectal cancer cells","volume":"50","author":"Lu","year":"2017","journal-title":"Int. J. Oncol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"115526","DOI":"10.18632\/oncotarget.23308","article-title":"Curcumol potentiates celecoxib-induced growth inhibition and apoptosis in human non-small cell lung cancer","volume":"8","author":"Cai","year":"2017","journal-title":"Oncotarget"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/24\/17610\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:40:54Z","timestamp":1760132454000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/24\/17610"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,18]]},"references-count":50,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["ijms242417610"],"URL":"https:\/\/doi.org\/10.3390\/ijms242417610","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,18]]}}}