苦参碱引起C6胶质瘤细胞凋亡及其对TRADD表达的影响

The Effects of Matrine in Apoptosis of C6 Glioma Cells and TRADD Expression

目的:应用一系列分子生物学检测技术来检测苦参碱对胶质瘤细胞中TRADD表达的影响,以进一步推测其诱导胶质瘤细胞凋亡的作用机制。方法:采用MTT法测定不同浓度苦参碱作用于C6胶质瘤细胞后的吸光值以计算其抑制率;采用Annex- in/FITC染色进行流式细胞术检测来测定不同浓度苦参碱诱导C6胶质瘤细胞的凋亡率;采用ICC(免疫细胞化学)、Western-blot- ting及Real-time PCR array(实时定量PCR芯片)方法来检测苦参碱对胶质瘤细胞TRADD表达的影响。结果:MTT结果显示细胞抑制率随着药物剂量的增加而增加;Annexin/FITC染色进行流式细胞术的检测结果显示胶质瘤细胞的凋亡率随着药物剂量的增加而增加;ICC、Western-bloning及Real-time PCR array结果显示苦参碱可诱导胶质瘤细胞TRADD基因及蛋白表达的上调。结论:苦参碱可以诱导胶质瘤细胞凋亡,其诱导胶质瘤细胞凋亡可能是通过上调TRADD的表达,以死亡受体途径来实现的。

Objective： To detect the effect of matrine on TRADD expression in C6 glioma cells by a series of molecular biological techniques and to study its mechanism of inducing C6 glioma cells＇ apoptosis. Methods： C6 glioma cells induced by different concentrations of matrine was tested by MTT. Apoptosis of C6 glioma cells was detected by FCM （Flow cytometry） （Annexin V/PI staining）; TRADD protein expression was detected by ICC （immunocytochemistry）, Western-blotting and real-time PCR array. Results： The detecting results showed that the growth inhibition ratio and apoptosis of the C6 cells increased with the enhancing of medicinal dosage and matrine could induce the up-regulation of TRADD expression. Conclusion： Matrine can induce C6 glioma cells＇ apoptosis, which may be related to the up-regulation of TRADD expression and the death receptors way.