柴胡皂苷D通过下调miR-517a调节胶质瘤细胞体外和体内移植瘤生长

目的探究柴胡皂苷D通过调控miR-517a对胶质瘤细胞体外和体内移植瘤生长的影响.方法采用MTT试剂盒检测U-87细胞和正常星形细胞的存活率;Q-PCR检测miR-517a表达;流式细胞仪检测细胞凋亡;Transwell实验检测细胞迁移和侵袭;CCK-8试剂盒检测细胞增殖;蛋白免疫印迹试剂盒检测cleaved caspase-3、Ki67、MMP-2、Cyclin D1蛋白水平.建立裸鼠移植瘤模型,检测各组肿瘤重量;免疫组化检测Ki67、VEGF表达.结果柴胡皂苷D对正常星形细胞存活率无影响,降低U-87细胞存活率,下调miR-517a表达,促进细胞凋亡,上调cleaved caspase-3蛋白表达,抑制细胞迁移、侵袭、增殖,下调MMP-2、Cyclin D1、Ki67蛋白表达;抑制体内移植瘤生长,降低Ki67、VEGF阳性表达率.结论柴胡皂苷D具有下调miR-517a调节胶质瘤细胞体外和体内移植瘤生长的作用.

Saikosaponin D Regulates Growth of Glioma Cells in vitro and Trans-planted Tumor in vivo by Down-regulating miR-517a

Objective To investigate the effect of Saikosaponin D on the growth of glioma cells in vitro and in vivo by regulating miR-517a.Methods Survival rates of U-87 cells and normal astro-cytes were determined by MTT kit.miR-517a expression was detected by q-PCR.Cell apoptosis was assessed by flow cytometry.Transwell assay was applied to detect cell migration and invasion.CCK-8 kit was used to detect cell proliferation.Cleaved caspase-3,Ki67,MMP-2 and Cyclin D1 protein levels were detected by Western blotting.The model of transplanted tumor in nude mice was estab-lished and the weight of tumor in each group was detected.The expression levels of Ki67 and VEGF were detected by immunohistochemistry.Results Saikosaponin D had no effect on the survival rate of normal astrocytes,decreased the survival rate of U-87 cells,down-regulated the expression of miR-517a,promoted apoptosis,up-regulated cleaved caspase-3 protein expression,inhibited cell migration,invasion and proliferation,and down-regulated MMP-2,Cyclin D1 and Ki67 protein expression.The growth of transplanted tumor was inhibited,and the positive expression rates of Ki67 and VEGF were de-creased by Saikosaponin D.Conclusion This study shows that Saikosaponin D can down-regulate miR-517a to regulate the growth of glioma cells in vitro and transplanted tumor in vivo.