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p15基因对人肝癌HepG2顺铂耐药细胞周期和耐药性的影响研究
引用本文:朱振双,陈伟庆,房殿亮,张杨.p15基因对人肝癌HepG2顺铂耐药细胞周期和耐药性的影响研究[J].中国药房,2012(41):3888-3890.
作者姓名:朱振双  陈伟庆  房殿亮  张杨
作者单位:重庆医科大学附属第二医院消化内科,重庆400010
摘    要:目的:利用p15-shRNA表达载体干扰细胞周期相关基因p15,探讨p15基因对人肝癌HepG2顺铂耐药细胞周期和耐药性的影响。方法:用梯度浓度的顺铂诱导HepG2细胞耐药并建立动态耐药模型HepG2/顺铂/2.0,以不同浓度(30、60、90nmol·L-1)的干扰质粒p15-shRNAY2和阴性对照质粒(shRNA-F)转染HepG2/顺铂/2.0细胞后48h检测p15荧光阳性细胞,筛选最佳干扰浓度;以筛选结果转染HepG2/顺铂/2.0细胞,与未转染细胞和阴性对照转染细胞比较,检测细胞存活情况、周期分布和p15、Bcl-2、Bax的蛋白表达。结果:最佳干扰浓度为60nmol·L-1;与未转染细胞和阴性对照转染细胞比较,p15-shRNAY2转染细胞的存活率明显增加,G1期细胞明显减少,p15、Bax蛋白表达明显降低,Bcl-2蛋白表达明显增加(P均<0.01)。结论:p15-shRNA可抑制p15基因表达,干扰细胞周期使G1期的比例减少,增加HepG2/顺铂/2.0对顺铂的耐药性。

关 键 词:细胞周期  p15基因  肝癌  顺铂耐药  shRNA干扰

Effects of p15 Gene on Cell Cycle and Drug Resistance of Human Hepatoma Cell Line HepG2 Resistant to Cisplatin
ZHU Zhen-shuang,CHEN Wei-qing,FANG Dian-liang,ZHANG Yang.Effects of p15 Gene on Cell Cycle and Drug Resistance of Human Hepatoma Cell Line HepG2 Resistant to Cisplatin[J].China Pharmacy,2012(41):3888-3890.
Authors:ZHU Zhen-shuang  CHEN Wei-qing  FANG Dian-liang  ZHANG Yang
Institution:(Dept. of Digestive Diseases, The Sec- ond Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China)
Abstract:OBJECTIVE: To interfering in cell cycle related gene p15 by pl5-shRNA expression vector, and to investigate the effect of p15 gene on cell cycle and drug resistance of human hepatoma cell line HepG2 resistant to cisplatin. METHODS: Drug re- sistance of HepG2 cell was induced by different concentrations of cisplatin and dynamic drug-resistance model HepG2/cisplatin/2.0 was established. HepG2/cisplatin/2.0 cell line was transfected with different concentrations (30, 60, 90 nmol. L-1) of interference plasmid (p15-shRNA Y2) and negative control interference plasmid (shRNA-F), and 48 h later the fluorescence positive cells were determined and the optimal interference concentration was screened. HepG2/cisplatin/2.0 cell line was transfected according to the re- suits of screening and compared with non-transfected cell and negative control transfected cell. The cell viability, cycle distribution and the protein expressions of plS, Bcl-2 and Bax were determined. RESULTS: The optimal interference concentrations was 60 nmol . L - 1. Compared with non-transfected cell and negative control transfected cell, the viability of transfected cell targeting plS-shRNAY2 increased significantly and the number of it decreased significantly at G1, phase; protein expression of p15 and Bax decreased and that of Bcl-2 increased significantly (all P〈0.01). CONCLUSION: p15-shRNA can inhibit the expression of p15 gene, and the fraction of G1 subpopulation is decreased by interfering in cell cycle and increased the cisplatin-resistance of HepG2/ cisplatin/2.0 to cisplatin.
Keywords:Cell cycle  p 15 gene  Liver cancer  Cisplatin-resistance  shRNA interference
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