摘要
[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。
[Purpose] To investigate the effect of Bcl-w overexpression on proliferation and sensi- tivity to cisplatin chemotherapy of bladder cancer cell line 5637. [Methods] The plasmid pcD- NA3.1（＋） was used to restructure plasmid pcDNA3.1-Bcl-w.Westeru blot was performed to confirm the effect of pcDNA3.1-Bcl-w after 5637 cells were transfected. The growth curve, survival curve, colony formation ability and cell cycle of pcDNA3.1-Bcl-w cells and control cells were detected by CCK-8, plate colony formation assay and flow cytometry,respectively. [ Results ] Plasmid pcD- NA3.1-Bcl-w was successfully constructured and transfected to 5637 cells. Compared with control cells,pcDNA3.1-Bclw ceils had a higher growth curve at the point of 24h,48h and 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 vs 1.520± 0.021, P〈0.001） ; formed more colonies （60.67±6.03 vs 48.67±6.11, P〈0.001） ; had a lower propor- tion of G0/GI phase （50.76% vs 57.02%,P〈0.001）;had a higher survival curve at different cisplatin concentration [ （83.8%±1.7%）vs （93.9%±1.2%） ,P〈0.001 at 30μmol/L; （59.5%±0.7%）vs （72.5%±1.9%） ,P=-0.028 at 50μmol/L; （50.5%±2.2%）vs（57.2%±1.2%）,P=0.007 at 70μmol/L; （37.2%±0.7%）vs （45.7%±1.4%）,P〈0.001 at 90p, mol/L; （29.1%±0.5%）vs （33.2%±0.6%）,P〈 0.001 at 110μmol/L]. [Conclusion ] The overexpression of Bcl-w faciliates the proliferation of bladder cancer cells 5637 and weakens the sensitivity of cancer cells to cisplatin chemotherapy.
出处
《肿瘤学杂志》
CAS
2014年第7期555-559,共5页
Journal of Chinese Oncology
基金
上海市科委基础研究项目(12ZR1417700)
