奥沙利铂诱导人肝癌细胞株HepG2凋亡及其机制探讨

Study on apoptosis of human hepatoma cell line HepG2 induced by oxaliplatin

目的 :研究奥沙利铂对人肝癌细胞株HepG2生长抑制及诱导凋亡的作用并探讨其诱导凋亡的作用机制。方法 :应用MTT法检测奥沙利铂对HepG2细胞生长的抑制作用 ;电镜观察细胞的形态改变 ;流式细胞仪分析细胞周期的分布和凋亡的情况 ;免疫组织化学法检测突变型p5 3,Bcl- 2 ,Bax ,Fas等凋亡相关基因蛋白的表达。结果 :奥沙利铂对HepG2有明显的抑制作用 ,细胞阻滞于S期和G2 /M期 ,G0 /G1期细胞减少 ,作用 72h出现凋亡峰 ,电镜可以找到凋亡小体 ,免疫组织化学表明Bax表达增强 ,突变型 p5 3、Bcl- 2表达减弱 ,Fas表达无差异。 结论 :奥沙利铂具有抑制肝癌细胞株HepG2增殖及诱导凋亡作用 ,其机制与促凋亡相关基因表达增强和抗凋亡相关基因表达减弱有关。

Objective:To study the effects of oxaliplatin on the proliferation and apoptosis of human hepatoma cell line HepG2 and to investigate its mechanism.Methods:The inhibition of HepG2 cell proliferation was estimated by MTT.Morphologic changes under electron microscope were observed.Alteration of cell cycle and apoptosis were analyzed using flow cytometry.The expression of apoptosis-associated genes mutant type p53,Bcl-2,Bax and Fas were detected with immunohistochemical technique.Results:Oxaliplatin could apparently inhibit the growth of HepG2 cells.Cell cycle analysis indicated that oxaliplatin blocked cells in S and G 2/M phase and the apoptosis emerged when treated with oxaliplatin for 72 hours.Under transmission electron microscope,apoptotic body could be found.The expression of genes Bax was up-regulated,the expression of genes mutant type p53,Bcl-2 was down-regulated,while the expression of Fas was not changed.Conclusion:Oxaliplatin can inhibit the proliferation and induce apoptosis of HepG2 cells.The apoptosis is related to the up-regulation apoptotic genes and down-regulation of anti-apoptotic genes.