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RNA干扰下调p53抑制因子iASPP表达对膀胱癌细胞的影响
引用本文:刘涛,李琳,贾麾,荆宏伟,孔垂泽.RNA干扰下调p53抑制因子iASPP表达对膀胱癌细胞的影响[J].中华泌尿外科杂志,2011,32(4).
作者姓名:刘涛  李琳  贾麾  荆宏伟  孔垂泽
作者单位:1. 中国医科大学附属第一医院泌尿外科,沈阳,110001
2. 广东省人民医院康复科
3. 中国医科大学
基金项目:辽宁省博士科研启动基金
摘    要:目的 探讨RNA干扰使p53抑制因子iASPP表达下调对膀胱癌细胞的影响.方法 iASPP siRNA慢病毒感染入膀胱癌细胞株5637和T24,实时定量PCR和蛋白质印迹法检测iASPP的表达;噻唑盐法测定细胞生长;集落形成测定法检测集落形成比率;荧光激活细胞分选术检测细胞周期.结果 iASPP siRNA慢病毒感染后细胞iASPP mRNA为0.60±0.02,低于阴性对照慢病毒组(1.00±0.03,P<0.05).膀胱癌细胞iASPP蛋白表达降低.iASPP下调能抑制入膀胱癌细胞5637和T24的生长和增殖(P<0.05),每个集落中细胞数显著减少(P<0.05),集落数目也低于阴性对照组(P<0.05).iASPP siRNA慢病毒5637细胞培养中的G1期细胞比率明显高于阴性对照慢病毒组,分别为(51.94±0.98)%和(46.00±0.77)%;T24细胞结果与之类似,分别为(60.04±0.45)%和(53.62±0.69)%;组间比较差异均有统计学意义(P<0.05).结论 RNA干扰能够使膀胱癌细胞iASPP表达下调,从而抑制膀胱癌细胞的生长和增殖.
Abstract:
Objective To discuss the effects of silencing of iASPP gene on human bladder cancer cells. Methods RNAi silencing of iASPP gene in bladder cancer cell 5637 and T24 cells were used by lentiviral mediated interfering short hairpin RNAs. Cell proliferation was tested by MTT assay, and rate of colony was tested by colony formation assay. Cell cycles were tested by using fluorescence-activated cell sorting. Results Down-regulation of iASPP could inhibit the growth and proliferation of human bladder cancer cells (P<0.05). iASPP know-down could decrease the colony formation of 5637 and T24 cells (P<0, 05). Knocking down of iASPP in 5637 and T24 cells showed cell arrested at G1. Conclusions Silencing of iASPP gene could inhibit proliferation and colony formation of bladder cancer, iASPP might be an important target for gene therapy of bladder cancer.

关 键 词:RNA干扰  肿瘤抑制蛋白质p53  膀胱肿瘤  

Effects of silencing of iASPP gene on human bladder cancer cells
LIU Tao,LI Lin,JIA Hui,JING Hong-wei,KONG Chui-ze.Effects of silencing of iASPP gene on human bladder cancer cells[J].Chinese Journal of Urology,2011,32(4).
Authors:LIU Tao  LI Lin  JIA Hui  JING Hong-wei  KONG Chui-ze
Abstract:Objective To discuss the effects of silencing of iASPP gene on human bladder cancer cells. Methods RNAi silencing of iASPP gene in bladder cancer cell 5637 and T24 cells were used by lentiviral mediated interfering short hairpin RNAs. Cell proliferation was tested by MTT assay, and rate of colony was tested by colony formation assay. Cell cycles were tested by using fluorescence-activated cell sorting. Results Down-regulation of iASPP could inhibit the growth and proliferation of human bladder cancer cells (P<0.05). iASPP know-down could decrease the colony formation of 5637 and T24 cells (P<0, 05). Knocking down of iASPP in 5637 and T24 cells showed cell arrested at G1. Conclusions Silencing of iASPP gene could inhibit proliferation and colony formation of bladder cancer, iASPP might be an important target for gene therapy of bladder cancer.
Keywords:iASPP
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