盐酸青藤碱诱导人肝癌细胞SMMC-7721凋亡及其死亡受体DR4、DR5表达的变化

摘要

目的：观察人肝癌细胞SMMC-7721凋亡过程中死亡受体DR4、DR5的表达，探讨盐酸青藤碱诱导人肝癌细胞凋亡的作用机制。方法体外培养人肝癌细胞SMMC-7721，经不同浓度盐酸青藤碱作用后，采用MTT法检测细胞增殖情况；Hoechst染色荧光显微镜观察细胞凋亡；用Western blotting方法检测人肝癌细胞DR4、DR5的表达。结果盐酸青藤碱对人肝癌细胞的生长有明显的抑制作用，并呈剂量、时间依赖性，不同浓度组之间差异均有统计学意义（ P＜0．05）；Hoechst33258染色可见凋亡细胞皱缩，核碎裂，也可见典型凋亡小体；在蛋白水平，盐酸青藤碱处理后SMMC-7721细胞死亡受体DR4、DR5的表达水平较未处理组显著增加（ P＜0．05）。结论盐酸青藤碱可抑制人肝癌细胞SMMC-7721的细胞增殖，促进其凋亡，此作用可能与细胞内死亡受体DR4和DR5表达上调有关。%Objective To investigate the expression levels of DR4 and DR5 on the process of apoptosis induced by Sinomenine hydrochloride in hepatocellular carcinoma ( HCC) SMMC-7721 cells.Methods MTT assay was used to de-termine the effect of Sinomenine hydrochloride on the growth of hepatocellular carcinoma SMMC-7721 cells.In vitro cul-tured human HCC SMMC-7721 cells:the effect of Sinomenine hydrochloride on the morphologic change induced by sino-menine hydrochloride were assessed by fluorescence microscope after Hoechst staining.SMMC-7721 cells were cultured for 48 hours with Sinomenine hydrochloride 2 mmol/L, then the effect of Sinomenine hydrochloride on death receptors ( DR4 and DR5) were detected by Western blotting.Results Sinomenine hydrochloride inhibited the growth of human HCC SMMC-7721 cells in a concentration-and time-dependent manner as determined by MTT assay.In vitro cultured human HCC SMMC-7721 cells, Sinomenine hydrochloride induced apoptosis by Hoechst staining; Sinomenine hydro-chloride significantly increased death receptors ( DR4 and DR5) protein expression in a concentration-dependent manner in human HCC SMMC-7721 cells.Conclusion Sinomenine hydrochloride can inhibit growth of human HCC SMMC-7721 cells through apoptotic induction; Sinomenine hydrochloride can increase the expression levels of death receptors ( DR4 and DR5) on the process of apoptosis induced by Sinomenine hydrochloride in HCC SMMC-7721 cells.