紫杉醇联合紫草素对U87脑胶质瘤细胞的作用及机制初探

The effects and mechanism involved in paclitaxel combined with shikonin on U87 glioma cells

目的:探讨紫杉醇联合紫草素通过协同作用杀伤人脑胶质瘤的作用。方法:将不同浓度的紫杉醇与紫草素单药及两种药物半量联合作用于胶质瘤细胞后,用CCK-8法检测细胞增殖抑制率,用划痕实验和Transwell法检测细胞生物学行为(迁移与侵袭)的影响,流式细胞技术检测细胞周期及凋亡率,Western-blot测定细胞外调节蛋白激酶(ERK)的蛋白表达。结果:紫杉醇与紫草素对胶质瘤细胞株U87细胞增殖具有抑制作用,且呈时间和剂量依赖性,并且发挥协同作用。紫杉醇与紫草素单独以及联合应用抑制U87细胞迁移与侵袭。两种药物联合应用促进U87细胞凋亡,紫草素组G1期的RNA含量增多,紫杉醇组及联合应用组的G2期RNA含量显著增高。Western-blot结果显示两者联合应用时,p-ERK蛋白表达降低。结论:与紫杉醇或紫草素单用组相比,两者半量联用通过协同作用能显著抑制U87胶质瘤细胞增殖、迁移、侵袭能力,促进细胞凋亡,阻滞细胞周期。紫杉醇与紫草素的协同效应机制与二者联合应用抑制p-ERK蛋白表达相关。

Objective:To investigate the synergistic effect of paclitaxel combined with shikonin on human glioma.Methods:U87 cells were treated with different concentrations of paclitaxel combined with shikonin.Cell proliferation inhibition rate was measured by using CCK-8.The cell biological behavior(migration and invasion)was detected by scratch test and transwell assay.Cell cycle and apoptosis rate were detected by flow cytometry.The expression of ERK(extracellular regulated protein kinases)was measured by Western blot.Results:Paclitaxel combined with shikonin could inhibit the proliferation of U87 glioma cells in a time-and dose-dependent manner,which also played a synergistic role.Paclitaxel and shikonin can be used alone and jointly could inhibit the migration and invasion of U87 glioma cells.The RNA content of the G 1 phase was significantly increased in shikonin group,the RNA content of the G 2 phase significantly increased in paclitaxel group and the combination group.The results of Western-blot showed that the expression of p-ERK protein was decreased in combination group.Conclusion:Compared with paclitaxel group and shikonin group,the combination group with half dosage can make synergic effects and significantly inhibit the proliferation,migration and invasion ability of U87 cells,besides promoting apoptosis and inhibiting cell circle.Paclitaxel combined with shikonin could decrease the expression of p-ERK/ERK,which may be involved in the synergic effects of these two drugs.