摘要
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 inducing effect of cantharidin in human lung cancer cells A549 and its molecular mechanisms. Methods MTT assay was used to determine A549 cells proliferation. Light and electron microscopy, FACScan, Annexin V FITC staining and DNA gel electrophoresis were used to detect apoptosis. The expression of bcl 2, Bax and survivin were examined by Western blot. Results Cantharidin inhibited the proliferation of A549 cells. The cells treated with cantharidin showed a typical apoptotic morphology and hypodiploid peak before G 1 phase. Flow cytometry analysis with annexin quantitatively further confirmed the increase of cell apoptosis. DNA of treated A549 cells depicted a ladder pattern characteristic of apoptosis, indicating the presence of DNA fragmentation. Western blot assay showed that cantharidin increased the level of Bax expression and inhibited the level of bcl 2 and survivin expression. Conclusion Cantharidin can induce A549 cells apoptosis mainly via regulation of Bax, bcl 2 and survivin expression.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2005年第6期330-334,共5页
Chinese Journal of Oncology
