miR-141靶向ZEB2对神经胶质瘤细胞增殖、迁移及侵袭的影响

任同; 杨智君; 陈思源; 黄磊

doi:10.13898/j.cnki.issn.1000-2200.2023.07.002

miR-141靶向ZEB2对神经胶质瘤细胞增殖、迁移及侵袭的影响

Effects of miR-141 targeting ZEB2 on the proliferation, migration, and invasion of glioma cells

摘要

摘要:
目的探讨miR-141靶向E盒结合锌指蛋白2(ZEB2)对神经胶质瘤细胞增殖、迁移及侵袭的影响。
方法体外培养人神经胶质瘤细胞系U87，对其转染并分为空白对照组(NG组)、阴性转染组(mimics-NC组)、过表达miR-141组(miR-141-mimics组)、共转染组(miR-141-mimics+pc-ZEB2组)。用实时荧光定量PCR法检测miR-141、ZEB2 mRNA水平；检测各组U87细胞增殖能力、迁移、侵袭能力；免疫印迹法检测增殖相关蛋白(PCNA)、侵袭及迁移相关蛋白(E-cadherin、N-Cadherin、Vimentin)表达情况；应用TargetScan数据库预测miR-141与ZEB2的靶向关系并用双荧光素酶报告基因实验加以验证。
结果U87细胞成功转染mimics-NC、miR-141-mimics及pc-ZEB2；与NG组、mimics-NC组比较，miR-141-mimics组U87细胞增殖抑制率、E-Cadherin蛋白表达升高(P < 0.05)，克隆形成率、划痕愈合率、侵袭细胞数、ZEB2 mRNA及其蛋白、PCNA、N-Cadherin、Vimentin蛋白表达降低或减少(P < 0.05)；与miR-141-mimics组比较，miR-141-mimics+pc-ZEB2组U87细胞增殖抑制率、E-Cadherin蛋白表达降低(P < 0.05)，克隆形成率、划痕愈合率、侵袭细胞数、ZEB2 mRNA及其蛋白、PCNA、N-Cadherin、Vimentin蛋白表达升高或增多(P < 0.05)。
结论过表达miR-141可能靶向下调ZEB2抑制神经胶质瘤U87细胞增殖、侵袭及迁移。

Abstract:
ObjectiveTo investigate the effects of miR-141 targeting zinc finger E-box binding homeobox 2 (ZEB2) on the proliferation, migration, and invasion of glioma cells.
MethodsHuman glioma cell line U87 was cultured in vitro, transfected, and then divided into blank control (NG) group, negative transfection (mimics-NC) group, and overexpression miR-141 (miR-141-mimics) group, co-transfection (miR-141-mimics+pc-ZEB2) group.Real-time fluorescence quantitative PCR was used to detect the levels of miR-141 and ZEB2 mRNA; the proliferation, migration and invasion of U87 cells in each group were detected; Western blotting was used to detect the expression of proliferation cell nuclear antigen (PCNA), invasion and migration-related proteins (E-cadherin, N-Cadherin, Vimentin); the targeting relationship between miR-141 and ZEB2 was predicted by TargetScan database and verified by dual luciferase reporter gene experiment.
ResultsU87 cells were successfully transfected with mimics-NC, miR-141-mimics, and pc-ZEB2;compared with the NG group and the mimics-NC group, the U87 cell proliferation inhibition rate and E-Cadherin protein expression in the miR-141-mimics group were significantly increased (P < 0.05), and the clone formation rate, scratch healing rate, number of invasive cells, the expression of ZEB2 mRNA and protein, and the expression of PCNA, N-Cadherin, and Vimentin protein were reduced (P < 0.05);compared with the miR-141-mimics group, the U87 cell proliferation inhibition rate and E-Cadherin protein expression in the miR-141-mimics+pc-ZEB2 group were significantly reduced (P < 0.05), and the clone formation rate, scratch healing rate, number of invasive cells, the expression of ZEB2 mRNA and protein, and the expression of PCNA, N-Cadherin, and Vimentin protein were increased (P < 0.05).
ConclusionsOverexpression of miR-141 may target and down-regulate ZEB2 to inhibit the proliferation, invasion, and migration of glioma U87 cells.

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