人肺腺癌细胞系GLC-82中p21^(WAF1)和p53基因的过表达

Overexpression of p21WAF1 and p53 in Human Lung Adenocarcinoma Cell Line

目的 探讨p21^(WAF1)和p53基因过表达对人肺腺癌细胞生长和细胞凋亡的影响。方法 用带有p21cDNA的真核表达载体pCEP和带有p53的腺病毒表达载体pAdCMV,分别通过基因转染和腺病毒感染,使其在人肺腺癌细胞系(GLC-82)中过表达;通过流式细胞仪、透射电镜和原位末端分析(TUNEL)观察细胞的生长和凋亡情况。结果 p21过表达的细胞系G1期细胞增加,细胞生长抑制,克隆形成率下降,电镜下未见特征性凋亡小体,原位细胞凋亡分析未见凋亡信号;p53腺病毒感染的肿瘤细胞增殖明显抑制并出现死亡,原位细胞凋亡检测到凋亡信号。结论 p21和p53基因的过表达能够抑制人肺腺癌细胞的生长,其中p53能够诱发细胞凋亡,因此这两种基因在肿瘤基因治疗上有着广泛的应用前景。

Objective To study the growth inhibitory effects of p21WAF1 and p53 overexpression in human lung adenocarcinoma cell line. Methods The p21WAF1 and p53 gene were transfected respectively into a human lung adenocarcinoma cell line, GLC-82. Flow cytometry (FLC) , transmission electron microscopy (EM) and TUNEL technique were used to evaluate cell growth and identify apoptosis. Results The GLC-82 transfected by p21 plasmid showed increased cell number in G1 phase of cell cycle, decreased proliferation potential and decreased cloning efficiency. Apoptosis have not been detected neither on EM nor by TUNEL technique, whereas the GLC-82 infected by Ad-p53 showed significantly decreased proliferation potential and some of them even died, in addition apoptosis was confirmed by TUNEL technique. Conclusion The results indicate that p21WAF1 and p53 can inhibit proliferation; p53 also can induce apoptosis of lung adenocarcinoma cell. Therefore, these two genes should have a wide application in gene therapy of tumors in future.