Zhao C, Ma Z G, Mou S L, Yang Y X, Zhang Y H, Yao W C
Department of Neurosurgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Department of Neurosurgery, The People's Hospital of Rizhao, Rizhao, Shandong, China.
Genet Mol Res. 2017 Mar 30;16(1):gmr-16-01-gmr.16019281. doi: 10.4238/gmr16019281.
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.
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。