Rab11对膀胱癌细胞顺铂化疗耐药性的影响及机制

Effect of Rab11 on cisplatin resistance in bladder cancer cells

目的探讨小鸟苷三磷酸(GTP)酶Rab11对膀胱癌细胞顺铂化疗耐药性的影响及作用机制。方法培养人膀胱癌细胞,分为观察1组、对照1组、观察2组、对照2组,分别加入2μmol/L顺铂诱导凋亡。24 h后,观察1组细胞转染Rab11特异性siRNA,对照1组细胞转染Control siRNA,观察2组细胞转染Rab11过表达质粒,对照2组细胞转染p CMV6质粒。转染48 h后收集各组细胞,采用CCK-8法测算细胞存活率,采用流式细胞术测算细胞凋亡率,采用Western blotting法检测细胞B淋巴细胞瘤-2(Bcl-2)蛋白表达。结果观察1组细胞存活率低于对照1组,观察2组细胞存活率高于对照2组(P均<0.05)。观察1组细胞凋亡率高于对照1组,观察2组细胞凋亡率低于对照2组(P均<0.05)。观察1组细胞Bcl-2蛋白表达低于对照1组,观察2组细胞Bcl-2蛋白表达高于对照2组(P均<0.05)。结论 Rab11能增强膀胱癌细胞对化疗药物顺铂的耐药性,其机制可能为调控凋亡相关蛋白Bcl-2的表达。

Objective To investigate the effect and mechanism of the small guanosine triphosphate enzyme（ GTPase） Rab11 on cisplatin resistance in bladder cancer cells. Methods The cultured human bladder cancer cells were divided into the observation group 1,control group 1,observation group 2,and control group 2. Apoptosis was induced by addition of 2 μmol/L cisplatin. After 24 h,cells were transfected with Rab11-specific siRNA in the observation group 1 and the cells were transfected with control siRNA in the control group 1. The cells were transfected with Rab11 overexpression plasmid in the observation group 2,and cells were transfected with p CMV6 plasmid in the control group 2. After 48-hour transfection,cells of each group were harvested,the cell viability was measured by CCK-8,apoptosis rate was determined by flow cytometry,and B-cell lymphoma 2（ Bcl-2） protein was detected by Western blotting. Results The survival rate of the cells in the observation group 1 was lower than that in the control group 1,and the survival rate of the cells in the observation group 2 was higher than that in the control group 2（ both P〈0. 05）. The apoptosis rate of the cells in the observation group 1 was higher than that in the control group1,and the apoptosis rate of the cells in the observation group 2 was lower than that in the control group 2（ both P〈0. 05）. The Bcl-2 protein expression in the observation group 1 was lower than that in the control group 1; the expression of Bcl-2 protein in the observation group 2 was higher than that in the control group 2（ both P〈0. 05）. Conclusion Rab11 can enhance the resistance of bladder cancer cells to cisplatin by regulating the expression of apoptosis-associated protein Bcl-2.