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目的:研究艰难梭菌毒素A(TcdA)对人胆管癌细胞株FRH-0201细胞凋亡影响及作用机制。方法:采用MTT法检测细胞增殖抑制率,荧光染色检测TcdA作用后细胞形态学变化,流式细胞术检测细胞凋亡,免疫细胞化学法检测Bcl-2表达水平,Caspase3活性检测试剂盒检测Caspase-3的活性。结果:TcdA能抑制胆管癌细胞株FRH-0201细胞增殖且呈时间、剂量依赖性,荧光染色和流式细胞仪检测到细胞凋亡,与对照组相比,TcdA作用后Bcl-2蛋白表达下降,差异有统计学意义(P<0.05),Caspase-3活性增强,差异有统计学意义(P<0.05)。结论:TcdA可以通过下调Bcl-2蛋白表达表达,激活Caspase-3而诱导FRH-0201细胞凋亡。
Aims: To study the effect of TCDA on the apoptosis of human cholangiocarcinoma cell line FRH-0201 and its mechanism. Methods: MTT assay was used to detect the inhibition rate of cell proliferation. Fluorescent staining was used to detect the morphological changes after TcdA treatment. Cell apoptosis was detected by flow cytometry. The expression of Bcl-2 was detected by immunocytochemistry. Caspase- 3 activity. Results: TcdA inhibited the proliferation of FRH-0201 cells in a time-and dose-dependent manner. Fluorescence staining and flow cytometry showed that the apoptosis of FRH-0201 cells was inhibited by TcdA. Compared with the control group, the expression of Bcl-2 protein decreased , The difference was statistically significant (P <0.05), Caspase-3 activity increased, the difference was statistically significant (P <0.05). Conclusion: TcdA can induce the apoptosis of FRH-0201 cells by down-regulating the expression of Bcl-2 protein and activating Caspase-3.