金丝桃苷通过miR-155/c-MYB通路抑制脑胶质瘤U251细胞增殖、侵袭及迁移作用

Effect of Hyperoside on inhibiting proliferation,invasion and migration of glioma U251 cells via miR-155/c-MYB pathway

目的探讨金丝桃苷通过miR-155/c-MYB通路抑制脑胶质瘤U251细胞增殖、侵袭及迁移的作用。方法将人神经胶质瘤细胞系U251分为U251细胞组、紫衫醇组(300μmol/mL)、金丝桃苷低剂量组(300μmol/mL)和金丝桃苷高剂量组(600μmol/mL),每组设置6个平行孔,继续培养72 h。细胞培养结束后,采用CCK-8法测定细胞增殖水平,Transwell法评估细胞迁移和侵袭能力,实时荧光定量聚合酶链反应(RT-qPCR)法及蛋白质印迹法测定miR-155、c-MYB mRNA和蛋白表达水平。结果与U251细胞组比较,紫衫醇组、金丝桃苷低、高剂量组吸光度(A)值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P<0.05);与紫衫醇组比较,金丝桃苷低剂量组A值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA蛋白表达水平升高,凋亡率降低,差异有统计学意义(P<0.05),而金丝桃苷高剂量组A值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P<0.05);与金丝桃苷低剂量组比较,金丝桃苷高剂量组A值、存活率、克隆形成数目、穿膜数目、miR-155、c-MYB mRNA和蛋白表达水平降低,凋亡率升高,差异有统计学意义(P<0.05)。结论金丝桃苷可降低胶质瘤U251细胞的活力、增殖和侵袭能力,并促进其凋亡,相关机制可能与金丝桃苷抑制miR-155、c-MYB表达进而抑制miR-155/c-MYB通路激活有关。

Objective To investigate the effect of Hyperoside on inhibiting proliferation,invasion and migration of glioma U251 cells via miR-155/c-MYB pathway.Methods Human glioma cell line U251 was divided into glioma U251 cell group,paclitaxel group(300μmol/mL),Hyperoside low-dose group(300μmol/mL)and Hyperoside high dose group(600μmol/mL),with 6 parallel pores in each group,and all the groups were cultured for 72 h.After cell culture,the cell proliferation level was determined by CCK-8 assay,cell migration and invasion ability was evaluated by Transwell assay,and the mRNA and protein expression levels of miR-155 and c-MYB were determined by RT-qPCR and Western blotting.Results Compared with U251 cell group,the A value,survival rate,number of clone formations,number of transmembrane cells,mRNA and protein expressions of miR-155 and c-MYB in paclitaxel group,Hyperoside low-dose group and Hyperoside high-dose group decreased,and the apoptosis rate increased,with statistical significance(P<0.05).Compared with paclitaxel group,the A value,survival rate,number of clone formation,number of transmembrane,mRNA and protein expressions of miR-155 and c-MYB in Hyperoside low-dose group increased,and the apoptosis rate decreased,with statistical significance(P<0.05).In Hyperoside high-dose group,the A value,survival rate,number of clone formation,number of transmembrane,mRNA and protein expressions of miR-155 and c-MYB decreased,and apoptosis rate increased,with statistical significance(P<0.05).Compared with Hypericin low-dose group.Compared with Hyperoside low-dose group,the A value,survival rate,number of clone formation,number of transmembrane,mRNA and protein expression of miR-155,c-MY B in the Hyperoside high-dose group decreased,the apoptosis rate increased,with statistical significance(P<0.05).Conclusion Hyperoside can reduce the viability,proliferation and invasion of glioma U251 cells,and promote their apopto sis.The mechanism may be related to Hyperoside inhibiting the expression of miR-155 and c-MYB,and thus inhibiting the activation of miR-155/c-MYB pathway.