摘要
目的 探讨NDGA诱导胃癌细胞凋亡的分子机制。方法 分光光度法检测caspase 3的细胞活性 ,同时用Westernblot分析caspase 3及其底物PARP。用Westernblot分析P5 3、caspase 3、PARP、P2 1wafl、P2 7kipl、Bcl 2和Bax的浓度。免疫法检测细胞色素C。结果 NDGA通过活化caspase 3诱导细胞凋亡 ,caspase 3的活化是由于细胞色素C从线粒体释放入细胞质而引起的。NDGA处理也可导致P2 7kipl、Bax蛋白水平升高 ,同时伴有细胞积聚于G1期。z DEVD fmk ,一种caspase 3抑制剂 ,可逆转caspase 3的活化及其引起的凋亡。结论 NDGA诱导的细胞凋亡是通过上调P2 7kipl、Bax的水平 ,释放细胞色素C ,最终活化caspase
Aim To explore the molecular mechanism involved in NDGA induced apoptosis.Method The activation of caspase 3 was measured by a colorimetric caspase 3 cellular activity and Western blotting of the cleavage of caspase 3and its substrate PARP.The concentration of P53,caspase 3,PARP,P21 wafl ,P27 kipl ,Bcl 2 and Bax was detected by Western blot analysis.Immunodetection was used to detect cytochrome c.Results NDGA induced apoptosis was preceded by the activation of caspase 3 which was due to the release of cytochrome c from mitochondria into cytosol.NDGA treatment also resulted in increased protein levels of P27 kipl and Bax.This was accompanied by accumulation of cells at G 1 phase.Activation of caspase 3 and subsequent apoptosis after NDGA treatment was reversed by z EDVD fmk,a specific caspase 3 inhibitor.Conclusions These results suggest that apoptosis indued by NDGA is mediated through up regulation of P27 kipl ,Bax,release of cytochrome c,and finally activation of caspase 3.
出处
《胃肠病学和肝病学杂志》
CAS
2001年第2期125-130,共6页
Chinese Journal of Gastroenterology and Hepatology
