土木香内酯对C6脑胶质瘤细胞迁移侵袭及凋亡的影响

摘要

目的 观察土木香内酯对大鼠C6胶质瘤细胞迁移侵袭及凋亡的影响及探讨可能的作用机制.方法 体外培养大鼠C6胶质细胞瘤株,采用四甲基偶氮唑盐(MTT)法检测土木香内酯(AL)对C6细胞活力的影响;通过划痕实验、Transwell小室实验检测AL对C6细胞迁移侵袭的影响;流式细胞仪检测AL对C6细胞诱导凋亡情况的影响,以及JC-1荧光探针检测AL对C6细胞线粒体膜电位变化的影响;采用免疫印迹(Western blot)方法检测经AL处理后,C6细胞中各相关蛋白的表达情况.结果 AL显著抑制C6细胞的增殖,且呈时间及剂量依赖性;AL处理24 h后,C6细胞的迁移侵袭能力明显受到抑制,N-cadherin蛋白显著下降而E-cadherin蛋白明显上升;凋亡细胞数明显上升而线粒体膜电位明显下降,细胞色素C/cleaved PARP/cleaved caspase-3/cleaved caspase-9蛋白均显著上调.结论 AL抑制C6细胞的作用机制,一方面可能是通过调节钙粘素(cadherin)蛋白抑制其迁移侵袭,另一方面可能通过调节细胞色素 C/caspase 信号通路诱导其凋亡发生.%Objective To investigate the inhibitory effect of alantolactone on glioma C6 cells and its possible mechanism. Methods Rat glioma cell line C6 cells were cultured in vitro. The effect of AL on the viability of C6 cells was detected by MTT assay. The effect of AL on migration and invasion of C6 cells was assessed by scratch test and Transwell chamber assay. Flow cytometry was used to detect the effect of AL on the induction of apoptosis in C6 cells, and the effects of AL on mitochondrial membrane potential in C6 cells was detected using JC-1 fluorescent probe. Western blot was used to detect the expression of related proteins in C6 cells after AL treatment. Results AL significantly inhibited the proliferation of C6 cells in a time-and dose-dependent manner. After AL treatment for 24 hours, the migration and invasion of C6 cells were significantly inhibited, N-cadherin protein was significantly decreased and E-cadherin protein was significantly increased, the number of apoptotic cells increased obviously while the mitochondrial membrane potential decreased significantly. The protein expressions of PARP/ cleaved caspase-3/cleaved caspase-9 were significantly up-regulated. Conclusions AL inhibits the proliferation of glioma C6 cells by inhibiting their migration and invasion through regulating the protein expression of cadherin and inducing apoptosis in C6 cells by regulating the cytochrome C/caspase signaling pathway.