外源性p21 waf1基因转染对人肺癌细胞系A549细胞增殖的影响

Effects of p21wafl gene transfection on the proliferation of human lung carcinoma cells

目的探讨外源性p21waf1基因转染对人肺癌细胞系A549细胞增殖的影响。方法用脂质体介导的方法将pIRES—p21waf1真核表达载体转染人肺癌细胞系A549分为pIRES—p21waf1转染组、pIRES—neo空载体组及未转染组，应用实时荧光定量RT-PCR、Western blot分别检测各组p21waf1基因mRNA及p21蛋白表达情况，MTT法检测细胞增殖情况，流式细胞仪分析细胞周期变化，所有数据进行统计学分析。结果p21waf1转染组p21waf1mRNA和p21蛋白高表达，差异有统计学意义（P〈0．01）；p21waf1转染组A549细胞生长速度明显低于空载体组和未转染组（P〈0．01）；流式细胞仪观察到p21waf1转染组细胞发生G1／S阻滞，G1期细胞比例显著高于空载体组和未转染组[（76．68±2．56）％vs（50．27±1．24）％、（49．89±1．31）％，P〈0．01）]，S期比例显著低于空载体组和未转染组[（12．48±1．08）％VS（39．59±1．19）％、（41．56±1．17）％，P〈0．01]，并出现凋亡峰。结论p21waf1基因转染可以抑制人肺癌细胞系A549细胞增殖并能诱导其发生细胞凋亡，可能为肺癌的基因治疗提供一条新的途径。

Objective To investigate the effect of transfected p21waf1 gene on the proliferation of human lung carcinoma cells. Methods p21waf1gene packaged with lipofectin was transfected into the cells of a human lung carcinoma cell line A549, which stably expressed p21waf1gene. The p21 wall mRNA and protein expression of A549-p21 waf1, and cells were detected by Real Time PC R and Western blot methods, respectively. The cell growth was detected by MTT. The cell cycle pattern was assayed by flow cytometry. Results After transfection of pIRES-p21waf1 the p21waf1 mRNA and protein of A549-p21waf1 were highly expressed as compared with other cells. Cell growth was obviously inhibited and resulted in an accumulation of cells in G1 , presenting that the proportion of the cells in G1 phase was obviously increased from （49.89 ± 1.31 ） % and （50.27 ± 1.24） % to （76. 68 ± 2.56） %, and that of the cells in S phases was obviously decreased from （39.59 ± 1.19） % and （41.56 ± 1.17 ） % to （ 12.48 ± 1.08 ） % respectively, as compared with control group. Conclusions p21 waft gene can inhibit the growth and induce apoptosis of human lung carcinoma cells, which could be a new method of gene therapy for the lung carcinoma in future.