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目的探讨含蟾酥胶囊血清诱导人肝癌BEL-7402细胞凋亡的作用机制。方法用中药血清药理学方法,不同浓度含药血清加入体外培养的人肝癌BEL-7402细胞,分别孵育24h、48h,显微镜下观察细胞形态学变化,MTT比色法检测细胞存活率,流式细胞仪检测细胞凋亡率,琼脂糖凝胶电泳测定DNA梯状条带,免疫细胞化学染色检测Bcl-2蛋白的表达。结果含蟾酥胶囊血清组部分细胞出现凋亡细胞形态学改变,细胞存活率降低,增殖受到抑制;流式细胞仪检测,可见明显的凋亡峰;孵育48h,琼酯糖凝胶电泳呈现梯状条带(DNALadder);免疫细胞化学检测显示Bcl-2表达明显降低。结论蟾酥胶囊可以诱导人肝癌细胞BEL-7402凋亡,其作用机制之一可能与下调Bcl-2蛋白的表达有关。
Objective To investigate the mechanism of apoptosis of BEL-7402 cells induced by serum containing Toad capsule. Methods Human hepatocellular carcinoma BEL-7402 cells cultured in vitro were cultured with different concentrations of serum containing traditional Chinese medicine. The cells were incubated for 24 and 48 hours respectively. Morphological changes were observed under a microscope. Cell viability was measured by MTT colorimetric assay. Flow cytometry The apoptotic rate of the cells was detected by flow cytometry. The DNA ladder was detected by agarose gel electrophoresis, and the expression of Bcl-2 protein was detected by immunocytochemical staining. Results Some cells in the group containing Chansu Capsule showed apoptotic morphological changes, cell survival rate decreased and proliferation inhibited. Flow cytometry showed obvious apoptotic peak. After 48 hours incubation, agarose gel electrophoresis showed ladder shape (DNALadder); Immunocytochemistry showed that Bcl-2 expression was significantly reduced. Conclusion Toad capsule can induce the apoptosis of human hepatocellular carcinoma cell line BEL-7402. One of its mechanisms may be related to the down-regulation of Bcl-2 protein expression.