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[目的]探讨过表达的Bcl-w蛋白对膀胱肿瘤细胞5637的增殖能力、顺铂敏感性的影响。[方法]采用质粒pcDNA3.1(+)构建Bcl-w过表达载体,转染5637细胞后用Western blot验证过表达效果,用CCK-8、平板克隆形成实验、流式细胞仪检测pcDNA3.1-Bcl-w细胞和对照组细胞的增殖和生存曲线、克隆形成能力、细胞周期。[结果]成功构建pcDNA3.1-Bclw重组质粒并转染5637细胞。pcDNA3.1-Bcl-w细胞增殖曲线在24、48、72h均高于对照组(0.814±0.022 vs 0.727±0.017,P=0.006;1.259±0.017 vs 1.078±0.027,P=0.001;1.787±0.024 vs1.520±0.021,P<0.001),差异具有统计学意义;克隆形成能力高于对照组(60.67±6.03 vs48.67±6.11,P<0.001),差异具有统计学意义;处于G0/G1期的比例低于对照组(50.76%vs57.02%,P<0.001),差异具有统计学意义;在30、50、70、90、110μmol/L各浓度顺铂处理下生存曲线高于对照组[(83.8%±1.7%)vs(93.9%±1.2%),P<0.001;(59.5%±0.7%)vs(72.5%±1.9%),P=0.028;(50.5%±2.2%)vs(57.2%±1.2%),P=0.007;(37.2%±0.7%)vs(45.7%±1.4%),P<0.001;(29.1%±0.5%)vs(33.2%±0.6%),P<0.001],差异具有统计学意义。[结论]高表达的Bcl-w增强了膀胱癌细胞5637的增殖能力,削弱了肿瘤细胞对化疗药物顺铂的敏感性。
[Objective] To investigate the effect of overexpression of Bcl-w protein on the proliferation and cisplatin sensitivity of bladder cancer cell line 5637. [Method] The Bcl-w overexpression vector was constructed by using the plasmid pcDNA3.1 (+). After transfected into 5637 cells, the over-expression was verified by Western blot. The expression of pcDNA3.1 was detected by flow cytometry with CCK-8, Bcl-w cells and control cells proliferation and survival curve, clonality, cell cycle. [Result] The pcDNA3.1-Bclw recombinant plasmid was successfully constructed and transfected into 5637 cells. The proliferation curve of pcDNA3.1-Bcl-w cells were higher than that of the control group at 24,48,72h (0.814 ± 0.022 vs 0.727 ± 0.017, P = 0.006; 1.259 ± 0.017 vs 1.078 ± 0.027, P = 0.001; 1.787 ± 0.024 vs1 .520 ± 0.021, P <0.001). The difference was statistically significant. The clonogenic capacity was higher than that of the control group (60.67 ± 6.03 vs 48.67 ± 6.11, P <0.001), and the difference was statistically significant. (50.76% vs57.02%, P <0.001), and the difference was statistically significant. The survival curves were higher in cisplatin treated groups of 30, 50, 70, 90 and 110μmol / L than those in control group [ P <0.001; (59.5% ± 0.7%) vs. (72.5% ± 1.9%), P = 0.028; (50.5% ± 2.2%) vs. (57.2% P <0.001); P = 0.007; (37.2% ± 0.7%) vs (45.7% ± 1.4% The difference was statistically significant. [Conclusion] Highly expressed Bcl-w enhances the proliferation of bladder cancer cell line 5637 and weakens the sensitivity of tumor cells to chemotherapeutic drug cisplatin.