p21基因对肝癌细胞的生物学影响

P21 gene transfection on the biologic behavior of hepatocellular carcinoma cells

目的探讨逆转录病毒介导的p21基因对肝癌Hep3B细胞的杀伤作用。方法通过脂质体将有p21基因的逆转录病毒载体MMLV-p21转染肝癌Hep3B细胞系,并用G418筛选。试验分为转p21基因的Hep3B-P21组及相同遗传背景及培养过程的两对照组肝癌Hep3B系细胞组、转Hep3B-MMLV组3组。采用RT-PCR检测实验组p21基因的表达;应用流式细胞计数分别分析3组细胞周期;MTT法检测细胞增殖并比较细胞抑制率。结果野生p21基因阻止Hep3B系细胞进入S期,停滞在G1期,Hep3B-P21组细胞增殖比两对照组明显受到抑制(P<0.05)。结论逆转录病毒介导p21基因可有效抑制肝癌Hep3B系细胞的增殖并诱导肝癌细胞的凋亡。

Objective To investigate the role of the retroviral vector inserted p21 gene in apoptosis of hepatomacells.Methods MMLV-p21,the retroviral vector inserted p21 gene was transfected into Hep3B cells by LipofectAMINE and selected in medium containing G418 for Hep3B,Hep3B-MMLV,Hep3B-p21 Cell lines for comparison were matched on the same genetic background and passage.The expression of p21 were detected by RT-PCR.Flow cytometry was used to analysis cell cycle.MTT assay was used to examine the proliferation inhibitory effect of p21 on Hep3B cell,and calculate the inhibitory rate of growth.Results The wild type p21 gene prevented the hepatoma cells from entering the S period,suspending in the G1 period.The proliferative ability of Hep3B-p21 was much more inhibited than in control（P＆lt;0.05）group.Conclusion Our data demonstrate that p21 can inhibit the proliferation of Hep3B cells and induce apoptosis of hepatocellular carcinoma cells.