Tizanidine对胶质瘤细胞U251增殖迁移及凋亡影响.docVIP

Tizanidine对胶质瘤细胞U251增殖迁移及凋亡影响 　　[摘要] 目的 探究Tizanidine对胶质瘤细胞U251的增殖、迁移及凋亡的影响，并尝试探讨其作用机制。 方法 将U251细胞分为两组，一组用Tizanidine（20 μmol/L）处理24 h；另一组用DMSO处理为对照组。采用CCK8实验检测细胞增殖情况；采用Transwell实验检测细胞的迁移和侵袭情况；采用细胞流式检测细胞凋亡情况，采用Western blot检测PI3K-AKT信号通路相关蛋白及凋亡相关蛋白的表达量的变化。 结果 CCK8增殖实验表明Tizanidine可以抑制脑胶质瘤细胞U251的增殖（P 0.05）；Transwell结果说明Tizanidine可以抑制脑胶质瘤细胞U251的迁移和侵袭（P 0.05）；流式凋亡结果分析显示经无血清凋亡诱导后，经Tizanidine处理的细胞凋亡明显增多（P 0.05），Western blot 结果显示经过Tizanidine处理的U251细胞，细胞的抗凋亡蛋白B淋巴细胞瘤-2（BCL-2）的表达量显著下调（P 0.05），促凋亡蛋白Bcl-2相关X蛋白（BAX）和Active Caspase3表达量则显著上调（P 0.05），AKT和mTOR的磷酸化水平均受到了显著抑制（P 0.05），PI3K/AKT信号通路的下游蛋白P70、CyclinD1的表达也相应下调（P 0.05）。 结论 Tizanidine抑制胶质瘤细胞U251的增殖、迁移和侵袭，诱导其凋亡，这有可能是通过抑制PI3K/AKT信号通路的激活实现的。 　　[关键词] Tizanidine；U251；PI3K/AKT信号通路 　　[中图分类号] Q71 [文献标识码] A [文章编号] 1673-7210（2018）01（b）-0027-05 　　[Abstract] Objective To investigate the effect of Tizanidine on the proliferation， migration and apoptosis of U251， and to explore its mechanism， as well. Methods U251 cells were divided into two groups， one group treated with Tizanidine （20 μmol/L）， and the other treated with DMSO as control group. CCK8 assay was used to detect cell proliferation； Transwell assay was performed to detect cell migration and invasion； flow cytometry was adopted to detect cell apoptosis； Western blot was used to detect the expression of PI3K-AKT signaling-related proteins and apoptosis-related proteins. Results The proliferation of U251 cell was inhibited by Tizanidine （P 0.05）； and Tizanidine treatment also inhibited migration and invasion of U251 cell （P 0.05）； Tizanidine induced apoptosis in U251 cell （P 0.05）； Western blot showed that the expression of Bcl-2 was significantly down-regulated （P 0.05）， and the expression of BAX and Active Caspase3 were both significantly up-regulated in Tizanidine-treated U251 cells （P 0.05）， respectively. In addition， Tizanidine significantly inhibited the phosphorylation levels of AKT and mTOR （P 0.05）， and the expression of related downstream proteins （P70s6k a