5-脱氧杂氮胞苷对人肝癌细胞HepG2增殖及beclin1表达的影响

Effect of 5-aza-2'-Deoxycity Dine on Proliferation of HepG2 Cells and Expression of Beclin1

目的:观察5-脱氧杂氮胞苷(5-aza-CdR)对HepG2细胞生长抑制及beclin1表达的影响,以探讨其抗肿瘤发生的潜在机制。方法:采用四甲基偶氮唑盐(MTT)法检测5-aza-CdR对HepG2细胞的生长抑制;用相差显微镜观察不同药物浓度下不同时间段的肝癌细胞形态学改变;采用RT-PCR法和Western blot法检测5-Aza-CdR对抑癌基因beclin1的mRNA和蛋白表达的影响。结果:5-aza-CdR可抑制HepG2细胞生长,呈剂量依赖性,并上调beclin1的mRNA和蛋白的表达。结果:显示102.4umol/L5-aza-C-dR作用72小时细胞增殖抑制率最高,可达(84.3±3.31)%,beclin1的mRNA和蛋白表达上调最明显,与对照组相比差异有统计学意义。结论:5-aza-CdR可抑制HepG2细胞增殖,其机制可能是通过恢复某些抑癌基因的表达,上调beclin1的mRNA和蛋白表达。

Objective： To observe the effect of 5-aza-2’-deoxycitydine on cell growth of HepG2 and the expression of beclin1,and to investigate the potential mechanism of its anti tumorigenesis.Methods： Cell growth inhibition was assayed by MTT method;Morphological changes of HepG2 were observed by phase contrast microscopy under different drug concentrations in different time periods;To detect the effect of 5-aza-CdR on expression of mRNA and protein in tumor suppressor gene beclin1 by RT-PCR and Western blot.Results： 5-aza-2’-deoxycitydine could inhibit HepG2 cell growth in a concentration-dependent manner,and increased beclin1 expression of mRNA and protein.The results showed that after 72 hours in 102.4umol/L5-aza-CdR intervention the rate of cell proliferation inhibition reached its maximum,up to（84.3±3.31）%,beclin1 expression of mRNA and protein increased most significantly.Difference was statistically significant compared with the control group.Conclusions： 5-aza-2’-deoxycitydine can inhibit the proliferation of HepG2 cells,and the mechanism may be that 5-aza-2’-deoxycitydine restores some tumor suppressor genes expression and increases the expression of beclin1.