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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 4’-methyl ether-scutellarein (4-MS) on apoptosis of JAR cells in human choriocarcinoma. Methods: The morphological and ultrastructural changes of Survivin, Caspase 9, Caspase 3 and p38 MAPK were detected by light microscopy and electron microscopy. The expressions of Survivin, Caspase 9, Caspase 3, p38 MAPK, Bax and cytochrome were detected by Western blot C protein expression differences. Results: Morphological changes of apoptotic cells were observed under light microscope and electron microscope. The results of RT-PCR showed that the expression of Survivin mRNA in JAR cells was significantly decreased after treated with 4-MS at different concentrations (20 and 40μg / ml) Caspase 9, Caspase 3 and p38 MAPK mRNA expression increased significantly. Western blot confirmed that Survivin protein expression in JAR cells was significantly decreased, the expression of Bax, cytochrome C protein and phosphorylation of p38 increased, and Caspase 3 and Caspase 9 were significantly activated. CONCLUSION: 4-MS can upregulate the expression of Bax, cytochrome C, Caspase 9 and Caspase 3, activate the p38 MAPK signal pathway and inhibit the expression of Survivin, and induce the apoptosis of JAR cells by regulating the expression of endogenous apoptosis signal transduction pathway Death.