摘要
目的 比较一个位点不同碱基的鼠p5 3minigene对细胞生长的影响。方法 三种小鼠p5 3mini gene[野生型 (Arg)、假野生突变型 (Leu)和突变型 (His) ]真核表达载体 ,由LipofectAMINE介导转染p5 3缺失的H12 99细胞 ,在转染后不同时间点收集细胞 ,利用细胞计数、流式细胞术、蛋白免疫印迹等方法比较三种p5 3对细胞体外生长的影响。结果 在转染后不同时间点 ,野生型和假野生突变型p5 3均能诱导细胞凋亡和周期阻滞 ,并可转录激活Bax、p2 1WAF1和Mdm 2 ,突变型则无以上效应。结论 野生型和假野生突变型p5 3具有诱导肺癌细胞凋亡和引起周期阻滞的功能 ,这种作用可能是通过转录激活Bax和p2 1WAF1而实现 ,Mdm 2与野生型和假野生型p5 3可以反馈调节。 172Leu和 172His两种突变表现出截然不同的生物学效应 ,支持p5 3基因存在保留野生型抑瘤功能突变方式的观点。
Objective To investigate the effect on cell growth after transfected with mouse 172 wild type p53 (Arg), pseudo wild type p53 (ArgLeu) and mutant p53 (ArgHis). Methods Vectors of three kinds of p53 minigene were transfected into p53 null H1299 cells through lipofectAMINE. Cells were collected at different time after transfection and analyzed by cell counting, flow cytometry and Western blotting. Results Wild type p53 and pseudo wild type p53 could induce cell apoptosis and cell cycle arrest at different time. Bax, p21 WAF1 and Mdm 2 were transactivated by wild type p53 and pseudo wild type p53. No tumor growth suppression was observed in mutant p53 transfected cells. Conclusion Wild type p53 and pseudo wild type p53 can inhibit cell growth by inducing apoptosis and cell cycle arrest through transactivation of Bax and p21 WAF1 . There exists feedback loop between Mdm 2 and p53. The different biological effect between 172Leu and 172His shows that some p53 variants such as 172 ArgLeu may act as wild type p53.
出处
《中国肺癌杂志》
CAS
2001年第2期94-98,共5页
Chinese Journal of Lung Cancer
