摘要
目的 :研究逆转录病毒介导的p2 7kip1基因过度表达对细胞周期和细胞增殖的影响。方法 :构建含人 p2 7kip1基因的逆转录病毒真核表达载体 ,应用脂质体介导法 ,将外源性 p2 7kip1基因转染人胃癌细胞系BGC 82 3,应用Westernblot、FCM等方法 ,检测p2 7kip1蛋白在胃癌细胞内的表达及其对细胞增殖的影响。结果 :获得含人 p2 7kip1cDNA的重组逆转录病毒载体 ,外源性野生型 p2 7kip1基因整合于胃癌细胞并获稳定表达 ,细胞生长速度受抑制 ,同时出现G1期阻滞。结论 :构建的 p2 7kip1重组逆转录病毒能有效介导外源性 p2 7kip1在人胃癌细胞系中高表达 ,抑制细胞增殖 ,为进一步研究以 p2 7kip1为目的基因的肿瘤基因治疗奠定了基础。
Objective:To study the inhibitory effects of retro virus mediated p27 kip1 gene on cell cycle and proliferation.Methods:Recombinant retrovirus eukaryotic expression vector pLNCX p27 kip1 containing exogenous human p27 kip1 cDNA was constructed and used to infect cells.The exogenous p27 kip1 expression in BGC 823 cells and its effects on proliferation were determined by Western blot,FCM staining.Results:Exogenous p27 kip1 had successfully been transferred into BGC 823 cells and obtained permanent expression.The growth rate of these transfected BGC 823 cells was inhibited.The percentage of phase G 1 cells was increased and that of phase S cells was decreased by analyzing cell cycle.Conclusion:p27 kip1 recombinant retrovirus prepared in this study can efficiently transfer p27 kip1 into gastric cell line,and the over expression of p27 kip1 inhibits the proliferation,suggesting that p27 kip1 might be used as a potential therapeutic agent for the gene therapy on human gastric cancer.
出处
《中国现代普通外科进展》
CAS
2002年第3期149-151,共3页
Chinese Journal of Current Advances in General Surgery
