微小RNA-138调控Wnt通路对体外人脑胶质瘤细胞生物学行为的影响

doi:10.16098/j.issn.0529-1356.2023.06.009

摘要/Abstract

摘要：

目的探讨微小RNA（miR）-138调控Wnt信号通路对脑胶质瘤细胞生物学行为的影响。方法选取脑胶质瘤细胞系U-87MG、U251，将其随机分为空白组、miR对照组（miR-NC）、miR-138模拟物组（miR-138 mimics）和miR-138 抑制剂组（miR-138 inhibitor）。采用Real-time PCR检测各组细胞中miR-138表达；采用MTT、流式细胞术、Transwell实验及划痕实验分别对各组细胞的增殖、凋亡、侵袭以及迁移能力进行检测，采用Western blotting检测各组细胞Wnt通路相关蛋白表达。结果 MiR-138 mimics组miR-138表达水平较其余3组高，miR-138 inhibitor组低于空白组和miR-NC组(P＜0.05)；与空白组相比，miR-138 mimics组细胞增殖率较低，miR-138 inhibitor组较高，且成时间依赖性（P＜0.05）；MiR-138 mimics组细胞凋亡率高于空白组、miR-NC组和miR-138 inhibitor组，而miR-138 inhibitor组细胞凋亡率均低于其余组（P＜0.05）；MiR-138 mimics组侵袭细胞数低于空白组、miR-NC组和miR-138 inhibitor组，而miR-138 inhibitor组均高于其余3组（P＜0.05）；MiR-138 mimics组细胞迁移率低于空白组、miR-NC组和miR-138 inhibitor组，而miR-138 inhibitor组均高于其余3组（P＜0.05）；MiR-138 mimics组Wnt3a、Wnt1、糖原合成激酶3β（GSK-3β）和β-连环蛋白（β-catenin）蛋白表达低于空白组、miR-NC组和miR-138 inhibitor组；而miR-138 inhibitor组均高于其余3组（P＜0.05）。结论 MiR-138过表达可有效抑制脑胶质瘤细胞增殖、侵袭和迁移并促进其凋亡，可能是通过抑制Wnt信号通路而实现。

关键词: 微小RNA-138, 脑胶质瘤, Wnt信号通路, 生物学行为, 免疫印迹法

Abstract:

Objective To investigate the effect of microRNA (miR)-138 regulation of Wnt signaling pathway on the biological behavior of human glioma cells in vitro. Methods Glioma cell lines U-87MG and U251 were selected and randomly divided into blank group, miR-NC group, miR-138 mimics group and miR-138 inhibitor group. Real-time PCR was used to detect the miR-138 expression in each group; MTT, flow cytometry, Transwell assay and scratch assay were used to detect proliferation, apoptosis, invasion and migration ability of each group respectively, and Western blotting was used to detect Wnt pathway related protein expression in each group. Results The miR-138 expression level was higher in the miR-138 mimics group compared with the remaining 3 groups, and that in the miR-138 inhibitor group was lower than that in the blank group and the miR-NC group (P<0.05); Compared with the blank group, the cell proliferation rate was lower in the miR-138 mimics group and higher in the miR-138 inhibitor group, and was time-dependent(P<0.05); The apoptosis rate in the miR-138 mimics group was higher than that in the blank group, miR-NC group, and miR-138 inhibitor group, while the apoptosis rate in the miR-138 inhibitor group was lower than that in the rest other groups(P<0.05); The number of cell-invading cells in the miR-138 mimics group was lower than that in the blank group, miR-NC group, and miR-138 inhibitor group, while all miR-138 inhibitor group were higher than the remaining three groups (P<0.05); The cell migration rate of miR-138 mimics group was lower than that of blank group, miR-NC group and miR-138 inhibitor group, while all miR-138 inhibitor group were higher than the remaining three groups (P<0.05); Wnt3a, Wnt1, glycogen synthase kinase 3β(GSK-3β) and β-catenin protein expression in the miR-138 mimics group was lower than that in the blank group, miR-NC group, and miR-138 inhibitor group; While miR-138 inhibitor groups were higher than the remaining three groups(P<0.05). Conclusion MiR-138 overexpression effectively inhibite the proliferation, invasion and migration of glioma cells and promote their apoptosis, probably achieved by pathway inhibition of the Wnt signaling pathway.

Key words: MicroRNA-138, Glioma, Wnt signaling pathway, Biological behavior, Western blotting

中图分类号:

R739.41

吴俊波杨杰肖锋李金亮 .

微小RNA-138调控Wnt通路对体外人脑胶质瘤细胞生物学行为的影响 [J]. 解剖学报, 2023, 54(6): 682-688.

WU Jun-bo YANG Jie XIAO Feng LI Jin-liang . Effect of microRNA-138 regulation of Wnt pathway on the biological behavior of human glioma cells in vitro[J]. Acta Anatomica Sinica, 2023, 54(6): 682-688.

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