转染野生型p53基因对人肺腺癌细胞作用的研究

Research on the Effects of p53 Gene in Human Lung Adenocarcinoma Cell Line

目的 研究外源性野生型 p5 3基因对人肺腺癌细胞系GLC 82的生物学作用。 方法 将含有野生型 p5 3基因的pDOR neo逆转录病毒载体 ,通过Lipofectin转染GLC 82细胞 ,采用流式细胞分析、电镜下观察、3 H TdR掺入试验、软琼脂培养集落形成率测试及裸小鼠成瘤性实验 ,观察野生型p5 3基因对GLC 82细胞的生物学效应。结果 电镜下观察可见 ,转染野生型p5 3基因可使GLC 82细胞出现一些病理现象 ,如细胞质中有明显的空泡及线粒体肿胀 ;流式细胞仪分析结果显示 ,野生型 p5 3基因可阻止GLC 82细胞周期停止于G1 S区域 ;3 H TdR掺入试验结果提示野生型 p5 3基因能够抑制GLC 82细胞的DNA合成 ,软琼脂培养集落形成率减少及裸小鼠成瘤减慢。结论 转染野生型p5 3基因可抑制GLC 82细胞恶性增殖。

Objective To determine the effect of p53 gene on human lung adenocarcinoma cell line GLC-82.Methods Retrovival vector pDOR-neo containing p53 gene was transferred into GLC-82 cells by lipofectin technique.The effect of wp53 gene on GLC-82 cells was analyzed by flow cytometry(FCM),electron microscope,~3H-TdR incorporation test,soft agar culture and tumorigenicity test.Results Some vacuoles and swollen mitochondria as the signs of cell pathological change were found under electron microscope in the cytoplasma of transferred GLC-82 cells.The FCM analysis showed that cell cycle of the transferred GLC-82 cells was blocked in G1-S phase.Synthesis of DNA in the transferred GLC-82 Cells was inhibited in ~3H-TdR incorporation test;the rate of colony-forming decreased in soft agar culture and the tumorigenicity in nude mice decreased.Conclusion The expression of p53 gene can inhibit the growth of GLC-82 cells and tumor formation in nude mice.