摘要
目的观察沙蟾毒精诱导人肝癌SMMC-7721细胞凋亡的作用。方法用MTT法观察沙蟾毒精对人肝癌SMMC-7721细胞增殖的影响;通过荧光染色、流式细胞分析等方法观察沙蟾毒精对细胞凋亡的影响;RT-PCR和Western blot分析沙蟾毒精对Bcl-2、Bax、p53和p21等基因和蛋白表达的影响。结果沙蟾毒精对人肝癌SMMC-7721细胞的生长有明显的抑制作用,24 h半数抑制剂量(IC50)为0.415μg/mL。细胞周期结果显示,人肝癌SMMC-7721细胞经不同浓度的沙蟾毒精处理后,G2/M期细胞数量增加,而G0/G1期细胞数明显减少。RT-PCR和Western blot结果显示,人肝癌SMMC-7721细胞经沙蟾毒精处理后,Bax的表达升高,而bcl-2的表达降低(P<0.05)。结论沙蟾毒精可能通过线粒体凋亡途径诱导人肝癌SMMC-7721细胞凋亡。
Objective： To investigate the apoptosis induced by Arenobufagin and its mechanism. Methods： The in- hibition rates were measured by MTT assay in human hepatocellular carcinoma cells SMMC-7721 treated Arenob- ufagin. Cell circle analysis experiments were performed to detect apoptosis induced by Arenobufagin.Additionally Real-time RT-PCR analysis and Western blot were performed to detect the expression of Bcl-2, Bax, p53 and p21. Results： Arenobufagin displayed the marked inhibition effect to SMMC-7721 cells and the IC50 value is 0.415 μg/ mL. The percentages of cells accumulated in the G2/M phase increased and G0/GI decreased treated with Arenobufa- gin. The expression of Bax significantly increased and the expression of Bcl-2 decreased compared to the control in all cells treated with Arenobufagin （P 〈 0.05）. Conclusion： Arenobufagin can inhibit the cellular proliferation via G2/M phase arrest in dose-dependent manner and induce apoptosis through mitochondria pathway.
出处
《中国中医急症》
2013年第11期1845-1847,1883,共4页
Journal of Emergency in Traditional Chinese Medicine
基金
陕西省教育厅专项科研计划项目(11JK0682)
