4'-甲醚-黄芩素诱导人绒毛膜癌细胞凋亡的信号转导机理研究

The signal transduction mechanism of induced apoptotic effects of 4′-methylether-scutellarein on human choriocarcinoma JAR cells

目的:探讨4'-甲醚-黄芩素(4'-methyl ether-scutellarein,4-MS)诱导人绒毛膜癌JAR细胞凋亡的信号传导机制。方法:光镜、电镜观察细胞形态、超微结构,RT-PCR技术检测Survivin、Caspase 9、Caspase 3、p38 MAPK基因的表达,Western blot检测Survivin、Caspase 9、Caspase 3、p38 MAPK、Bax和cytochrome C蛋白水平的表达差异。结果:光镜和电镜下可见凋亡细胞的形态学改变;RT-PCR结果显示,不同质量浓度(20、40μg/ml)的4-MS作用24h后,JAR细胞中Survivin mRNA表达显著下降,而Caspase 9、Caspase 3及p38 MAPK mRNA表达显著上升。Western blot证实,JAR细胞的Survivin蛋白表达量显著下降,Bax、cytochrome C蛋白表达量及p38磷酸化水平升高,Caspase 3与Caspase 9被明显激活。结论:4-MS通过上调Bax、cytochrome C、Caspase 9、Caspase 3表达,激活p38 MAPK信号通路及抑制Survivin表达,对JAR细胞内源性细胞凋亡信号转导通路进行正向调节而诱导细胞凋亡。

Objective ： To investigate the signal transduction mechanism of apoptosis on human choriocarcinoma JAR cell line affected by 4＇-methery-scutellarein（4-MS）.Methods： Cell morphology and ultrastructure was examined by a light and electronic microscope.mRNA expressions of survivin、Caspase 9、Caspase 3、p38 MAPK JAR cells upon 4＇-M-S treatment were examined via Real-Time quantitative PCR.Western blot was employed to detected the expression of Survivin、Caspase 9、Caspase 3、p38 MAPK、Bax and cytochrome C after the treatment 4-MS.Results ： The morphological changes of apoptosis appeared on JAR cells treated by 4-MS;It had been observed by RT-PCR that the survivin mRNA expression decreased and the Caspase 9,Caspase 3,phosphorylated p38 mRNA expression increased on the JAR cells treated by 4-MS in different concentration（20 and 40μg/ml） for 24h（P0.05）.Western blot analyses showed that protein expression of survivin was decreased,but Bax,cytochrome C,phosphorylated p38,Caspase 3 and Caspase 9 were extremely up-regulated by 20μg/ml and 40μg/ml 4-MS treatment.Conclusions： 4-MS can induce JAR cell apoptosis both by down-regulation of Survivin,and the direct activation of Bax,cytochrome C,Caspase 3,Caspase 9 and p38 MAPK,which have an active effection on endogenous apoptosis signal transduction pathway.