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目的探讨斑蝥素诱导人肺癌A549细胞的凋亡作用及其分子机制。方法采用MTT法检测斑蝥素对A549细胞的增殖抑制作用;以光镜、电镜、流式细胞仪、AnnexinV2FITC标记法和DNA凝胶电泳检测细胞凋亡;以蛋白印迹法分析斑蝥素对bcl22、Bax和Survivin蛋白表达的影响。结果斑蝥素能抑制A549细胞的增殖;斑蝥素处理A549细胞后,光镜与电镜下可见到明显的凋亡细胞;细胞周期的G1期前有低于二倍体细胞的凋亡峰;AnnexinV2FITC标记法的定量检测进一步证实了斑蝥素诱导细胞凋亡的作用;DNA凝胶电泳显示出典型的凋亡细胞特征;蛋白印迹检测表明斑蝥素可使Bax表达升高,bcl22和Survivin表达下降。结论斑蝥素能显著抑制A549细胞增殖,诱导细胞凋亡,并主要通过调节Bax、bcl22和Survivin等蛋白的表达来实现。
Objective To investigate the apoptosis of human lung cancer A549 cells induced by cantharidin and its molecular mechanism. Methods MTT assay was used to detect the inhibitory effect of cantharidin on A549 cells. The apoptosis of A549 cells was detected by light microscopy, electron microscopy, flow cytometry, AnnexinV2FITC and DNA gel electrophoresis. The expressions of bcl22, Bax And Survivin protein expression. Results Canthaxanthin inhibited the proliferation of A549 cells. After treatment with cantharidin, the apoptotic cells were found under light and electron microscopes. The apoptotic peak of the cell cycle was lower than that of diploid cells before G1 phase. AnnexinV2FITC Quantitative detection of this method further confirmed the role of cantharidin in inducing apoptosis. DNA gel electrophoresis showed the typical characteristics of apoptotic cells. Western blotting showed that cantharidin increased the expression of Bax and decreased the expression of bcl22 and Survivin. Conclusion Canthaxanthin can significantly inhibit the proliferation and induce the apoptosis of A549 cells and mainly through the regulation of the expression of Bax, bcl22 and Survivin proteins.