Graduate Institute of Pathology and Parasitology, National Defense Medical Center, Taipei, R.O.C.
Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, R.O.C.
Kaohsiung J Med Sci. 2021 Feb;37(2):128-135. doi: 10.1002/kjm2.12306. Epub 2020 Oct 14.
The aim of the study is to investigate the ability of phytochemicals to overcome the multiple drug resistance (MDR) of bladder cancer. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the cytotoxic sensitivity of T24-GCB cells, a GCB resistant cell line, to different phytochemicals, including capsaicin, quercetin, curcumin, and resveratrol, and their combination with gemcitabine. Western blot analysis was used to detect the expression of membranous ABCC2 and metabolic proteins, DCK, TK1, and TK2 in tumor cells. Animal models were used to confirm the treatment efficacy of phytochemicals in combination with gemcitabine to bladder cancer. The observed/expected ratio of cytotoxicity analysis revealed that capsaicin has synergistic effect with gemcitabine to T24-GCB cells in a dose-dependent pattern. Quercetin, curcumin, and resveratrol have additive effect with gemcitabine to T24-GCB cells. Capsaicin and quercetin alone and combination with gemcitabine decreased the expression of ABCC2 and DCK and TKs, in T24-GCB cells. On the contrary, resveratrol and curcumin alone and combination with gemcitabine increased the expression of ABCC2 but decreased cytoplasmic kinases simultaneously. In xenografted subcutaneous tumor model on nude mice, combination treatment of capsaicin and gemcitabine demonstrated the highest tumor suppression effect when compared to capsaicin or gemcitabine treatment alone. The MDR of bladder cancer is closely related to membranous ABCC2, cytoplasmic DCK, and TKs expression. Capsaicin owns the strongest synergistic cytotoxic effect of gemcitabine to T24-GCB cells. This combination regimen may provide as an adjunctive treatment for overcoming MDR in bladder cancer.
本研究旨在探讨植物化学物质克服膀胱癌多药耐药（MDR）的能力。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法评估 GCB 耐药细胞系 T24-GCB 对不同植物化学物质（包括辣椒素、槲皮素、姜黄素和白藜芦醇）及其与吉西他滨联合的细胞毒性敏感性。Western blot 分析用于检测肿瘤细胞中膜 ABCC2 和代谢蛋白 DCK、TK1 和 TK2 的表达。动物模型用于证实植物化学物质与吉西他滨联合治疗膀胱癌的疗效。细胞毒性分析的观察/预期比值显示，辣椒素与吉西他滨对 T24-GCB 细胞具有剂量依赖性的协同作用。槲皮素、姜黄素和白藜芦醇与吉西他滨对 T24-GCB 细胞具有相加作用。辣椒素和槲皮素单独及与吉西他滨联合降低了 T24-GCB 细胞中 ABCC2 和 DCK 和 TKs 的表达。相反，白藜芦醇和姜黄素单独及与吉西他滨联合增加了 ABCC2 的表达，但同时降低了细胞质激酶的表达。在裸鼠皮下移植瘤模型中，与辣椒素或吉西他滨单独治疗相比，辣椒素和吉西他滨联合治疗显示出最高的肿瘤抑制效果。膀胱癌的 MDR 与膜 ABCC2、细胞质 DCK 和 TKs 的表达密切相关。辣椒素对 T24-GCB 细胞具有最强的与吉西他滨协同细胞毒性作用。这种联合治疗方案可能为克服膀胱癌的 MDR 提供辅助治疗。
