NDGA诱导胃癌细胞凋亡的分子机制的研究

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Western blot分析caspase-3及其底物PARP。用Western blot分析P53、caspase-3、PARP、P21^wafl、P27^kipl、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P27^kipl、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P27^kipl、Bak的水平，释放细胞色素C，最终活化caspase-3而引起的。

NDGA-induced apoptosis in gastric cancer cells mediated by up-regalatin p27 and activation of caopase-3

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.