槲皮素对人脑胶质瘤干细胞生物学行为及miR-29s家族的影响分析

The influence of quercetin on biological behavior of human glioma stem cells and the miR-29s family

目的探讨槲皮素对人脑胶质瘤干细胞生物学行为及微小RNA-29s(miR-29s)家族的影响。方法采用悬浮细胞培养人脑胶质瘤SHG44干细胞。根据槲皮素浓度不同分为0μg/ml(对照组),6μg/ml组、12μg/ml组、24μg/ml组、48μg/ml组、96μg/ml组,采用MTT法检测人脑胶质瘤SHG44增殖活性。用0μg/ml、12μg/ml、24μg/ml、48μg/ml培养48 h,采用细胞迁移实验和细胞侵袭实验检测细胞迁移数目和细胞侵袭数目;采用RT-PCR检测miR-29s家族表达。结果培养24 h和48 h,槲皮素各组抑制率高于对照组,且随着浓度增加抑制率不断增加,差异均有统计学意义(P <0. 05);培养48 h,槲皮素各组抑制率高于培养24 h,差异均有统计学意义(P <0. 05)。与对照组比较,槲皮素各组细胞迁移数目和细胞侵袭数目减少,差异均有统计学意义(P <0. 05);槲皮素48μg/ml组细胞迁移数目和细胞侵袭数目少于12μg/ml组和24μg/ml组,且24μg/ml组少于12μg/ml组,差异均有统计学意义(P <0. 05)。与对照组比较,槲皮素各组miR-29a和miR-29b表达升高,差异均有统计学意义(P <0. 05);槲皮素48μg/ml组miR-29a和miR-29b表达高于12μg/ml组和24μg/ml组,且24μg/ml组高于12μg/ml组,差异均有统计学意义(P <0.05)。结论槲皮素对人脑胶质瘤干细胞SHG44具有良好抑制作用,且具有剂量依赖性,降低体外迁移及侵袭能力,及促进miR-29a和miR-29b表达。

Objective To investigate the influence of quercetin on the biological behavior of human glioma stem cells and the microRNA-29 s family.Methods Human glioma SHG44 stem cells were cultured in suspension cells.Quercetin was divided into 0μg/ml(control group),6μg/ml,12μg/ml,24μg/ml,48μg/ml and 96μg/ml according to the concentration of quercetin.The proliferation activity of human glioma SHG44 was detected by MTT assay;the number of cell migration and cell invasion were detected by cell migration and cell invasion experiments;the expression of microRNA-29 s family was detected by RT-PCR.Results The inhibition rate of quercetin in each group was higher than that in the control group at 24 h and 48 h,and the inhibition rate increased with the increase of concentration(P<0.05).At 48 h,the inhibition rate of quercetin in each group was higher than that in 24 h(P<0.05).Compared with the control group,the number of cell migration and invasion in each group of quercetin decreased significantly(P<0.05).The number of cell migration and invasion in the 48μg/ml group was less than that in the 12μg/ml group and the 24μg/ml group,and the 24μg/ml group was less than that in the 12μg/ml group(P<0.05).Compared with the control group,the expression of microRNA-29 a and microRNA-29 b in quercetin group increased significantly(P<0.05).The expression of microRNA-29 a and microRNA-29 b in quercetin 48μg/ml group was higher than that in 12μg/ml group and 24μg/ml group,and the expression of microRNA-29 b in quercetin 24μg/ml group was higher than that in 12μg/ml group(P<0.05).Conclusion Quercetin has a good inhibitory effect on human glioma stem cells SHG44 in a dose-dependent manner,which reduces migration and invasion in vitro,and promotes the expression of microRNA-29 a and microRNA-29 b.