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  

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

作　　者：王冬芮[1] 张永慈 王增光[1] Wang Dongrui;Zhang Yongci;Wang Zengguang(Department of Neurosurgery,General Hospital of Tianjin Medical University,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院神经外科,300052

出　　处：《中华细胞与干细胞杂志（电子版）》2018年第3期129-133,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的探讨分析槲皮素对人脑胶质瘤干细胞(BGSCs)生物学行为及miR-29s家族的影响。方法使用干细胞培养液对U87人脑胶质瘤细胞进行培养,采用CCK-8法检测槲皮素对BGSCs细胞增殖抑制率,采用流式细胞技术检测槲皮素对BGSCs细胞凋亡影响,并采用real-time PCR鉴定槲皮素对BGSCs细胞中miR-29a、miR-29b以及miR-29c表达的影响。采用t检验以及方差分析进行统计学分析。结果随着槲皮素浓度的增加,BGSCs细胞增殖抑制率增加24 h 0μg/ml(0.00±0.12)﹪、10μg/ml(1.36±0.38)﹪、20μg/ml(15.33±3.01)﹪、40μg/ml(29.50±4.57)﹪、80μg/ml(40.21±6.42)﹪、160μg/ml(61.21±7.48),F=76.273,P <0.05;48 h 0μg/ml (0.09±0.05)﹪、10μg/ml(9.84±2.17)﹪、20μg/ml(28.57±3.84)﹪、40μg/ml(43.59±5.21)﹪、80μg/ml(59.50±3.28)﹪、160μg/ml(70.21±9.48)﹪,F=85.392,P <0.05,且同浓度槲皮素作用48 h对BGSCs细胞增殖抑制率高于作用24 h(P <0.05)。随着槲皮素浓度的升高,BGSCs细胞凋亡率升高[0μg/ml(13.42±1.21)﹪、20μg/ml(38.47±9.28)﹪、40μg/ml(59.34±7.20)﹪、80μg/ml(71.42±9.47)﹪,F=57.493,P <0.05]。不同浓度槲皮素处理BGSCs细胞后,可促进BGSCs细胞miR-29s家族miR-29a/b/c相对表达量,且随着槲皮素浓度的增加,BGSCs细胞miR-29s家族相对表达量增加[miR-29a 0μg/ml(1.04±0.08)、20μg/ml(1.16±0.05)、40μg/ml(1.30±0.10)、80μg/ml(1.41±0.09),F=19.281,P <0.05;miR-29b 0μg/ml(1.06±0.09)、20μg/ml(1.13±0.05)、40μg/ml(1.25±0.07)、80μg/ml(1.30±0.09),F=13.427,P <0.05;mi R-29c 0μg/ml(1.03±0.07)、20μg/ml(1.15±0.03)、40μg/ml(1.22±0.06)、80μg/ml(1.31±0.08),F=14.502,P <0.05]。结论槲皮素可有效抑制人脑BGSCs增殖,促进人脑BGSCs凋亡,并促进人脑BGSCs中miR-29s家族表达。Objective The effects of quercetin on the biological behavior of human glioma stem cells and the miR-29s family were investigated and analyzed. Methods The stem cell culture medium on U87 glioma cells were cultured to detect the inhibition rate of quercetin on the proliferation of BGSCs cells by CCK-8 method, and were detected by flow cytometry of quercetin on the apoptosis of BGSCs cells, the effect of real-time PCR on miR-29a miR-29b and the identification of quercetin and miR-29c expression in BGSCs cells. Results With the increase of quercetin concentration, the proliferation inhibition rate of BGSCs cells increased significantly [24 h 0 μg/ml（0.00±0.12）﹪, 10 μg/ml（1.36±0.38）﹪, 20 μg/ml（15.33±3.01）﹪, 40 μg/ml（29.50±4.57）﹪, 80 μg/ml（40.21±6.42）﹪, 160 μg/ml（61.21±7.48）,F = 76.273, P 〈 0.05; 48 h 0 μg/ml （0.09±0.05）﹪, 10 μg/ml（9.84±2.17）﹪, 20 μg/ml（28.57±3.84）﹪, 40 μg/ml（43.59±5.21）﹪,80 μg/ ml（59.50±3.28）﹪, 160 μg/ml（70.21±9.48）﹪, F = 85.392,P 〈 0.05]. The inhibition rate of 48 h on BGSCs cells was significantly higher than that of 24 h （P 〈 0.05）. With the increase of quercetin concentration, the apoptosis rate of BGSCs cells increased significantly [0 μg/ml （13.42±1.21）﹪, 20 μg/ml （38.47±9.28）﹪, 40 μg/ml （59.34±7.20）﹪, 80 μg/ml （71.42±9.47）﹪, F = 57.493, P 〈 0.05]. Quercetin treatment of different concentrations of BGSCs cells can promoted the relative expression of miR-29s family miR-29a/b/c in BGSCs cells. With the increase of quercetin concentration, the relative expression of miR-29s family in BGSCs cells increased significantly [miR-29a 0 μg/ml （1.04±0.08）, 20 μg/ml （1.16±0.05）, 40 μg/ml （1.30±0.10）、 80 μg/ ml（1.41±0.09）, F = 19.281, P 〈 0.05; miR-29b 0 μg/ml （1.06±0.09） , 20 μg/ml （1.13±0.05） , 40 μg/ml （1.25±0.07） , 80 μg/ml （1.30±0.09） , F = 13.427, P 〈 0.05; miR-29c 0 μg/ml （1.03±0.07） , 20 μg/ml （1.15±

关 键 词：槲皮素 人脑胶质瘤干细胞 生物学行为 微RNAS 

分 类 号：R285.5[医药卫生—中药学]