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微小RNA对CD133的靶向作用及其对胶质瘤细胞增殖和侵袭的影响分析

Targeting effect of microRNA on CD133 and its impact analysis on proliferation and invasion of glioma cells.

作者信息

Zhao C, Ma Z G, Mou S L, Yang Y X, Zhang Y H, Yao W C

机构信息

Department of Neurosurgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.

Department of Neurosurgery, The People's Hospital of Rizhao, Rizhao, Shandong, China.

出版信息

Genet Mol Res. 2017 Mar 30;16(1):gmr-16-01-gmr.16019281. doi: 10.4238/gmr16019281.

Abstract

MiR-200b, a member of the microRNA-200 family, has been identified to be capable of suppressing glioma cell growth through targeting CREB1 or CD133. However, whether miR-200b affects the biological behavior (proliferation, invasion, and migration) of glioma cells is poorly understood. The aim of this study was to evaluate the effect of miR-200b on the biological behavior of glioma cells in vitro. MiRNA-200b mimics, miRNA-200b inhibitor, and mimic control were transfected into conventionally cultured glioma U251 cells, followed by measuring the expression of miR-200b and CD133 in transfected cells by RT-PCR; effect of miR-200b on CD133 mRNA 3'-UTR luciferase activity by luciferase reporter assay; proliferation activity of transfected U251 cells by MTT method; and changes in U251 cell invasion and migration by Transwell method after transfection. Compared to that in the miRNA-200b inhibitor, mimic control, and blank control groups, miRNA-200b expression was significantly increased and CD133 mRNA expression was significantly decreased in the mimic miRNA-200b group in a time-dependent manner (P < 0.05). Meanwhile, dual luciferase reporter assay showed that miR-200b could inhibit CD133 activity through binding to the 3'-UTR of CD133 mRNA (P < 0.05). Furthermore, the proliferation activity and invasion and migration abilities of U251 cells transfected with miRNA-200b mimic were significantly decreased (P < 0.05). In conclusion, overexpression of miR-200b inhibited the proliferation, invasion, and migration of glioma cells possibly through targeting CD133.

摘要

MiR-200b是微小RNA-200家族的成员之一,已被证实能够通过靶向CREB1或CD133来抑制胶质瘤细胞的生长。然而,miR-200b是否影响胶质瘤细胞的生物学行为(增殖、侵袭和迁移)仍知之甚少。本研究的目的是评估miR-200b在体外对胶质瘤细胞生物学行为的影响。将miRNA-200b模拟物、miRNA-200b抑制剂和模拟对照转染到常规培养的胶质瘤U251细胞中,随后通过RT-PCR检测转染细胞中miR-200b和CD133的表达;通过荧光素酶报告基因检测法检测miR-200b对CD133 mRNA 3'-UTR荧光素酶活性的影响;通过MTT法检测转染后U251细胞的增殖活性;通过Transwell法检测转染后U251细胞侵袭和迁移的变化。与miRNA-200b抑制剂组、模拟对照组和空白对照组相比,模拟miRNA-200b组中miR-200b表达显著增加,CD133 mRNA表达随时间显著降低(P<0.05)。同时,双荧光素酶报告基因检测显示miR-200b可通过与CD133 mRNA的3'-UTR结合来抑制CD133活性(P<0.05)。此外,转染miRNA-200b模拟物的U251细胞的增殖活性以及侵袭和迁移能力显著降低(P<0.05)。总之,miR-200b的过表达可能通过靶向CD133抑制胶质瘤细胞的增殖、侵袭和迁移。

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