黑果枸杞花青素对人肝癌HepG2细胞凋亡与自噬相互作用的影响

Effects of Anthocyanins from Lycium ruthenicum Murr on the interaction between apoptosis and autophagy of human hepatoma HepG2 cells

为了研究黑果枸杞花青素(Anthocyanins from Lycium ruthenicum Murr,ALR)对人肝癌HepG2细胞凋亡与自噬相互作用的影响。将ALR与3-MA(自噬抑制剂)结合,采用Hoechst染色、流式细胞术、qRT-PCR和Western Blot法,测定人肝癌HepG2细胞核形态、凋亡率和凋亡因子(包括JAK2、STAT3、p-JAK2、p-STAT3)的mRNA表达及蛋白表达情况。将ALR与AG490(凋亡通路抑制剂)结合,使用qRT-PCR及Western Blot法检测人肝癌HepG2细胞自噬因子(LC3、LC3-Ⅱ)的mRNA表达及蛋白表达情况。结果表明:ALR结合3-MA后可诱导人肝癌HepG2细胞核出现明显的凋亡形态变化,提高人肝癌HepG2细胞凋亡率,显著抑制JAK2、STAT3的mRNA表达(P<0.05),显著抑制p-JAK2、p-STAT3的蛋白表达(P<0.05);ALR结合AG490后可显著提高LC3的mRNA表达及LC3-Ⅱ的蛋白表达(P<0.05)。综上所述,抑制细胞自噬可促进ALR诱导的人肝癌HepG2细胞凋亡,促进人肝癌HepG2细胞凋亡可增强细胞自噬。

Anthocyanins from Lycium ruthenicum Murr(ALR)is combined with 3-m A(autophagy inhibitor)in order to study the effects of ALR on the interaction between apoptosis and autophagy of human hepatoma HepG2 cells.The nuclear morphology,apoptosis rate,mRNA expression and protein expression of apoptosis factors(JAK2,STAT3,p-JAK2,p-STAT3)of human hepatoma HepG2cells are measured by Hoechst staining,flow cytometry,qRT-PCR and Western Blot.Combining ALR with AG490(apoptosis pathway inhibitor),mRNA expression and protein expression of autophagy factors(LC3,LC3-Ⅱ)in human hepatoma HepG2 cells are detected by qRT-PCR and Western Blot.The results show that the combination of ALR and 3-m A can induce significant morphological changes of apoptosis in human hepatoma HepG2 nucleus and improve the apoptosis rate of human hepatoma HepGI cells,so as to greatly inhibit the mRNA expression of JAK2 and STAT3(P<0.05)as well as the protein expression of p-JAK2 and p-STAT3(P<0.05).The combination of ALR and AG490 can significantly increase the mRNA expression of LC3 and the protein expression of LC3-Ⅱ(P<0.05).In conclusion,the inhibition of autophagy can promote the apoptosis of human hepatoma HepG2 cells induced by ALR.Promoting the apoptosis of human hepatoma HepG2 cells can improve autophagy.