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目的探讨p53基因和PTEN基因在脑胶质瘤细胞系U251发生发展过程中的作用机制。方法用不同MOI的p53腺病毒表达载体pAdCMV-p53及空载体pAdCMV-lacZ分别感染表达野生型PTEN基因和突变型PTEN基因的细胞系,RT-PCR及Westernblot方法检测转染效率;并通过MTT检测生长抑制率、流式细胞仪检测细胞周期及TUNEL检测分析细胞凋亡等指标观察p53基因及PTEN基因对U251细胞生长的影响。结果MOI为100时,p53基因可引起U251细胞G0G1期阻滞、诱导细胞凋亡,生长抑制;MOI为50时,U251-p53+PTEN生长抑制率明显高于U251-p53,并能出现细胞凋亡,而U251-p53仅出现少量细胞凋亡。结论p53基因可以通过细胞周期G0G1期阻滞及诱导细胞凋亡抑制胶质瘤细胞系U251的生长;PTEN基因可以促进p53基因对胶质瘤细胞系U251的生长抑制作用,并能增加U251细胞对p53基因诱导凋亡的敏感性。
Objective To investigate the mechanism of p53 gene and PTEN gene in the development of glioma cell line U251. Methods The transfected cells were transfected with pAdCMV-p53 and pAdCMV-lacZ at different MOI levels respectively. The transfection efficiency was detected by RT-PCR and Western blot. The expression of PTEN gene and mutant PTEN gene were detected by MTT assay Growth inhibition rate, cell cycle detected by flow cytometry and TUNEL detection of apoptosis and other indicators observed p53 gene and PTEN gene on the growth of U251 cells. Results When the MOI was 100, the p53 gene could block the G0G1 phase in U251 cells and induce apoptosis and growth inhibition. When the MOI was 50, the growth inhibition rate of U251-p53 + PTEN was significantly higher than that of U251-p53 However, U251-p53 showed only a small amount of apoptosis. Conclusion The p53 gene can inhibit the growth of glioma cell line U251 through cell cycle G0G1 arrest and apoptosis induction. PTEN gene can promote the growth inhibition of glioma cell line U251 by p53 gene and increase the ratio of U251 cells p53 gene induces apoptosis sensitivity.