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目的探讨斑蝥酸钠联合多西紫杉醇抑制人肺腺癌A549细胞增殖、诱导细胞凋亡的作用,初步探讨其作用机制。方法采用MTT法测定细胞增殖抑制率,Annexin V-FITC/PI染色,流式细胞仪测定细胞凋亡率;Western印迹法测定Caspase-8活性。结果斑蝥酸钠和多西紫杉醇均能抑制人肺腺癌A549细胞的增殖(P<0.05),合用与单用相比抑制率明显提高,诱导12 h时CI值为0.88,24 h时为0.96,36 h时为1.02。Annexin V-FITC/PI染色流式细胞仪检测提示斑蝥酸钠可增强多西紫杉醇的凋亡诱导作用。Western印迹法显示多西紫杉醇、斑蝥酸钠及联合加药组均可见凋亡相关蛋白Caspase-8的裂解条带,联合加药组的裂解条带最明显。结论多西紫杉醇和斑蝥酸钠均可明显抑制人肺腺A549细胞增殖诱导细胞凋亡,联合使用作用更强,多西紫杉醇和斑蝥酸钠诱导细胞凋亡可能与促进Caspases-8表达有关。
Objective To investigate the effect of sodium cantharidate combined with docetaxel on the proliferation and apoptosis of human lung adenocarcinoma A549 cells and to explore its mechanism. Methods The inhibitory rate of cell proliferation was determined by MTT assay. Annexin V-FITC / PI staining and flow cytometry were used to determine the apoptosis rate. Caspase-8 activity was determined by Western blotting. Results Both sodium cantharidinate and docetaxel could inhibit the proliferation of human lung adenocarcinoma A549 cells (P <0.05). Compared with the single use alone, the inhibition rate was significantly increased. The CI value at 12 h was 0.88 and 0.96 at 24 h , 1.02 at 36 h. Annexin V-FITC / PI staining revealed that sodium calendan could enhance the apoptosis-inducing effect of docetaxel. Western blotting showed that the cleavage bands of apoptosis-related protein Caspase-8 were found in docetaxel, sodium cantharidate and the combined dosing group. The cleavage band of the combined dosing group was the most obvious. Conclusions Both docetaxel and cantharidin sodium can significantly inhibit the proliferation of human lung adenocarcinoma A549 cells induced apoptosis, combined effect is stronger, docetaxel and cantharidate induced apoptosis may be related to the promotion of Caspases-8 expression.