p27^(KIP1)基因过表达诱导人胃癌细胞的凋亡

p27^(KIP1) Overexpression Causes Apoptosis in Human Gastric Carcinoma Cells

目的:研究p27KIP1基因对胃癌细胞凋亡的潜在调节作用。方法:采用脂质体转染法将p27KIP1全长cDNA转入胃癌细胞系SGC7901中,通过免疫印迹及RNA斑点杂交检测p27KIP1基因在蛋白质和mRNA水平的表达;通过DNA电泳、TUNEL染色、流式细胞仪及透射电镜等方法,观察目的基因对细胞凋亡的影响。结果:转染p27KIP1的SGC7901细胞在mRNA和蛋白质水平均有高水平p27KIP1的表达。诱导p27KIP1表达后,SGC7901细胞出现凋亡细胞所拥有典型的梯状DNA和超微结构改变,其凋亡指数也显著增加(P<0.01),并且p27KIP1的过表达使G1期前出现一个亚二倍体的凋亡峰,占14.68%。结论:p27KIP1基因能够诱导胃癌SGC7901细胞的凋亡。

Objective:To elucidate the effect of p27 KIP1 on apoptosis regulation in gastric carcinoma cells.Methods :We transfected the whole length of p27 KIP1 cDNA into human gastric cancer cells SGC7901by the method of lipofectin transfection.Expression of p27 KIP1 in protein or mRNA level were analyzed by Western blotting and RNA bot blotting respectively.DNA eletrophoresis,TUNEL,Flow cytometry and electron microscope were applied to assess the effect of p27 KIP1 on apoptosis.Results:Expression of p27 KIP1 in protein or mRNA increased evidently in SGC7901cells transfected with p27 KIP1 .p27 KIP1 overexpression induced apoptotic cell death reflected by pre -G 1 peak in the histogram of FACS,which was also confirmed by DNA eletrophoresis,TUNEL assay and electron microscope.Conclusion:The results indicate p27 KIP1 can induce apoptosis in gastric carcinoma.