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目的探讨槲皮素能否逆转膀胱癌细胞对顺铂的耐药性并研究其机制。方法 MTT法检测顺铂耐药T24膀胱癌细胞在顺铂和槲皮素处理下的细胞活力。流式细胞实验检测顺铂耐药T24膀胱癌细胞在顺铂和槲皮素处理下的细胞凋亡。Western blot试验检测顺铂耐药T24膀胱癌细胞在顺铂和槲皮素处理下Bim、活化caspase-9、活化caspase-3的表达水平以及细胞色素C、Smac/DIABLO的释放。免疫共沉淀实验检测顺铂耐药T24膀胱癌细胞在顺铂和槲皮素处理下Bim蛋白与Bak、Bax蛋白的相互作用。结果相比于常规T24细胞,顺铂耐药T24细胞对顺铂的敏感性显著下降。MTT和流式细胞实验结果表明槲皮素能显著促进顺铂对耐药T24细胞的杀伤活性和凋亡诱导活性。免疫共沉淀和Western blot试验结果表明槲皮素能显著上调顺铂耐药T24细胞中Bim蛋白的表达,从而通过与Bak、Bax蛋白的相互作用促进顺铂对肿瘤细胞线粒体膜孔道的开放,诱导细胞色素C和Smac/DIABLO从线粒体中释放到细胞质中,最终引起caspase-9和caspase-3的活化。结论槲皮素通过Bim-Bak/Bax途径提高顺铂耐药膀胱癌细胞对顺铂的敏感性。
Objective To investigate whether quercetin can reverse the drug resistance of bladder cancer cells to cisplatin and to study its mechanism. Methods MTT assay was used to detect cell viability of cisplatin-resistant T24 bladder cancer cells treated with cisplatin and quercetin. Flow cytometry was used to detect the apoptosis of cisplatin-resistant T24 bladder cancer cells treated with cisplatin and quercetin. Western blot was used to detect the expression of Bim, activated caspase-9, activated caspase-3 and the release of cytochrome C, Smac / DIABLO in cisplatin-resistant T24 bladder cancer cells treated with cisplatin and quercetin. Immunoprecipitation assay was used to detect the interaction between Bim protein and Bak, Bax protein in cisplatin - resistant T24 bladder cancer cells treated with cisplatin and quercetin. Results Compared with conventional T24 cells, the sensitivity of cisplatin-resistant T24 cells to cisplatin was significantly decreased. MTT and flow cytometry results show that quercetin can significantly promote cisplatin on drug-resistant T24 cell killing activity and apoptosis-inducing activity. Co-immunoprecipitation and Western blot showed that quercetin significantly up-regulated the expression of Bim protein in cisplatin-resistant T24 cells and promoted the opening of cisplatin to the mitochondrial membrane pore of tumor cells through the interaction with Bak and Bax proteins Cytochrome C and Smac / DIABLO are released from the mitochondria into the cytoplasm, eventually leading to the activation of caspase-9 and caspase-3. Conclusion Quercetin increases the sensitivity of cisplatin-resistant bladder cancer cells to cisplatin through the Bim-Bak / Bax pathway.