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目的:研究牡荆素(vitexin)对人肝癌细胞SMMC-7721的增殖抑制作用,并初步探讨其作用机制。方法:体外培养人肝癌细胞SMMC-7721,分别采用MTT法和Hoechst33258核染色法检测牡荆素对人肝癌细胞SMMC-7721活力的影响以及观察细胞形态学变化;流式细胞仪检测细胞凋亡率和线粒体膜电位(ΔΨm)变化;蛋白免疫印迹法检测p53、Bcl-2、Bax等相关凋亡蛋白水平的表达情况。结果:牡荆素培养人肝癌细胞SMMC-7721 48,72,96 h后能明显抑制细胞增殖,呈时间-剂量依赖性(P<0.05),其IC50分别是150.37,116.24,90.19μmol·L-1。牡荆素作用于人肝癌细胞SMMC-7721 72 h后,以浓度依赖性方式增加细胞凋亡率、降低线粒体膜电位(ΔΨm)以及上调p53、Bax、Caspase-3等相关促凋亡蛋白的表达,下调Bcl-2抗凋亡蛋白的表达。结论:牡荆素能抑制人肝癌细胞SMMC-7721增殖诱导凋亡,呈时间-剂量依赖性,其作用机制可能通过依赖P53途径下调Bcl-2,上调Casepase-3、Bax、P53、PARP等基因表达,进而诱导凋亡有关。
OBJECTIVE: To study the inhibitory effect of vitexin on the proliferation of human hepatocellular carcinoma cell line SMMC-7721 and to explore its mechanism. Methods: Human hepatocellular carcinoma cell line SMMC-7721 was cultured in vitro. MTT and Hoechst33258 nuclear staining were used to detect the effect of vitexin on the viability of human hepatocellular carcinoma cell line SMMC-7721. Morphological changes were observed by flow cytometry. And mitochondrial membrane potential (ΔΨm). Western blotting was used to detect the expression of p53, Bcl-2, Bax and other apoptosis proteins. Results: Vitexin cultured SMMC-7721 cells 48 h, 72 h, 96 h after injection significantly inhibited cell proliferation in a time-and dose-dependent manner (P <0.05). IC50 values were 150.37, 116.24 and 90.19 μmol·L- 1. Vitexin treated human hepatocellular carcinoma cells SMMC-7721 for 72 h, and increased the apoptosis rate, decreased mitochondrial membrane potential (ΔΨm) and up-regulated the expression of p53, Bax, Caspase-3 and other apoptosis proteins in a concentration-dependent manner , Down-regulated the expression of Bcl-2 anti-apoptotic protein. Conclusion: Vitexin inhibits the proliferation-induced apoptosis of human hepatocellular carcinoma cell line SMMC-7721 in a time-and dose-dependent manner. The mechanism may be through down-regulation of Bcl-2, upregulation of Casepase-3, Bax, P53 and PARP, And then induce apoptosis related.