含蟾酥胶囊血清诱导人肝癌BEL-7402细胞株凋亡的机制

Mechanism of Chansu capsule with medicated serum in inducing apoptosis of human hepatocellular carcinoma BEL-7402 cells

目的：探讨含蟾酥胶囊血清诱导人肝癌BEL-7402细胞株凋亡的机制。方法：用血清药理学方法，把含不同浓度蟾酥胶囊的血清加入体外培养的人肝癌BEL-7402细胞，分别孵育24、48h，显微镜下观察细胞形态学改变；MTT比色检测细胞的存活率；流式细胞仪检测细胞的凋亡率；琼脂糖凝胶电泳测定DNA条带；免疫细胞化学显色检测Bcl-2蛋白的表达。结果：实验组部分细胞凋亡，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，可见凋亡峰；孵育48h，琼脂糖凝胶电泳呈梯状条带（DNALadder）；免疫细胞化学检测，Bcl-2表达明显降低。结论：蟾酥胶囊诱导人肝癌细胞BEL-7402株凋亡的作用机制可能与下调Bcl-2蛋白表达有关。

Objective： To explore the mechanism of Chansu capsule with medicated serum in inducing the apoptosis of human hepatocellular carcinoma （HCC） BEL-7402 cells. Methods： Using serum pharmacologic method,we chose different concentrations of serum containing Chansu capsule, which were respectively co-cultivated with HCC BEL-7402 cells for 24 or 48 hours. The morphological change of apoptotic HCC cells was observed under a light microscope. The cell growth was examined by methyl-tetrazolium （MTT） assay. The percentage of apoptotic cells was examined by flow cytometry. DNA ladder was examined by agar gel electrophoresis. The expression of Bcl-2 protein was measured with immunocytochemical staining. Results： Some cell apoptosis appeared in the experimental groups and morphology remarkably changed. The cell growth was remarkably inhibited by the medicated serum contaning Chansu capsule. The apoptotic cell peaks were observed by flow cytomeric assays. DNA Ladder appeared at co-culturing 48 hours, and the expression of Bcl-2 protein decreased. Conclusion： The mechanism of Chansu capsule with medicated serum in inducing the apoptosis of human HCC BEL-7402 cells may be due to a decrease in protein expression of the Bcl-2 protein.