肉苁蓉苯乙醇苷对HepG2肝癌细胞增殖和凋亡的影响

Effect of Cistache deserticola phenylethanol glycoside on proliferaion and apoptosis of HepG2 cells in vitro

目的探讨肉苁蓉苯乙醇苷(Cistache deserticola phenylethanoid glycosides,CPhGs)对HepG2细胞增殖、凋亡的影响。方法实验以HepG2细胞为研究对象,采用MTT及平板克隆实验检测细胞增殖情况、Annexin V-FITC/PI、Hochest33258和线粒体染色法分析细胞凋亡、流式细胞术分析细胞周期,免疫印迹法检测Bcl-2、Cleaved-Caspase3及PARP表达情况。结果CPhGs可抑制HepG2细胞增殖,IC50为187μg/mL;平板克隆实验发现,CPhGs中、高剂量组对HepG2细胞有明显的抑制作用(P<0.01)。通过倒置显微镜、细胞核及线粒体染色发现,CPhGs各组均可诱导HepG2细胞发生凋亡。同时,Annexin V/PI双染实验结果表明,CPhGs各组均诱导HepG2细胞发生早期凋亡(P<0.01),且呈现一定的剂量依赖性;CPhGs各剂量组还可将HepG2细胞周期阻滞在S期(P<0.01)。Western-blot结果显示,CPhGs可诱到凋亡相关蛋白Cleaved-Caspase3及PARP表达的增加,同时还可抑制抗凋亡蛋白Bcl-2的表达(P<0.01)。结论CPhGs对HepG2细胞具有一定的抑制增殖、诱导凋亡的作用,其机制可能与调控细胞周期、诱导细胞凋亡相关蛋白表达有关。

Objective To investigate the effects of Cistache deserticola phenylethanol glycoside(CPhGs)on the proliferation and apoptosis of HepG2 cells.Methods Influence of CPhGs on HepG2 cell proliferation was measured by MTT and colony method.Apoptosis was detected by Annexin V-FITC/PI staining,Hoechst33258 staining and Mitotracker Red staining method.Cell cycle was determined by flow cytometry.The protein expression of apoptosis including Bcl 2,Caspase3 and PARP were detected by Western-blotting.Results The results showed that CPhGs could significantly inhibited proliferation of HepG2 cell(P<0.01),and IC50 value was 187μg/mL.Pro-apoptotic effect of CPhGs on HepG2 cell were revealed.Hoechst33258 staining found that the degree of nuclear appeared shrinking and fragmentation was dose-dependence of CPhG.Compared with control group,CPhGs could significantly induce HepG2 cell apoptosis with Annexin V/PI double staining experiment dose-dependently(P<0.01).Moreover,CPhGs could inhibit HepG2 cell cycle in S phase at a dose-effect dependent relation(P<0.01).In addition,Western-blotting results showed that CPhGs can increase the expression of Caspase3 and PARP,and decrease the expression of Bcl-2(P<0.05).Conclusion CPhGs can induce cell apoptosis by inhibiting cell proliferation,arresting cell cycle,and increasing apoptosis-related proteins.