论文部分内容阅读
该文目的为探讨miR-218-5p对表达A2B5+/CD133–的胶质瘤干细胞(glioma stem cells,GSCs)亚群干性维持及侵袭性的影响。实时荧光定量PCR(q RT-PCR)检测人脑胶质瘤组织及细胞株中miR-218-5p的表达;上、下调miR-218-5p慢病毒转染表达A2B5+/CD133–的胶质瘤干细胞SHG-139s,二次球体形成实验、免疫荧光及q RT-PCR检测SHG-139s的标志物A2B5、CD133、Nestin、PLAGL2、ALDH1及SOX2的表达变化;Transwell实验、q RT-PCR及免疫荧光检测SHG-139s基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的表达变化。结果显示,miR-218-5p在胶质瘤中低表达,上调miR-218-5p可显著降低SHG-139s成球率,同时明显降低SHG-139s中上述干细胞标志物的表达;Transwell实验显示,上调miR-218-5p能显著抑制SHG-139s的侵袭能力,使其MMP9表达受到明显抑制。因此,miR-218-5p可能作为抑癌基因在胶质瘤中发挥作用,高表达的miR-218-5p可抑制SHG-139s的干性维持及其侵袭能力。
The aim of this study was to investigate the effect of miR-218-5p on the maintenance and invasiveness of A2B5 + / CD133-expressing glioma stem cells (GSCs) subsets. Real-time quantitative PCR (q RT-PCR) was used to detect the expression of miR-218-5p in human glioma tissues and cell lines; miR-218-5p lentivirus was transfected into A2B5 + / CD133- The changes of SHG-139s markers such as A2B5, CD133, Nestin, PLAGL2, ALDH1 and SOX2 were detected by immunofluorescence and q RT-PCR. Transwell assay, q RT-PCR and immunofluorescence To detect the expression of matrix metalloproteinase 9 (MMP9) in SHG-139s. The results showed that miR-218-5p was down-regulated in gliomas. Up-regulation of miR-218-5p significantly decreased the rate of SHG-139s formation and decreased the expression of SHG-139s. Transwell assay showed that miR- Up-regulation of miR-218-5p significantly inhibited the invasion ability of SHG-139s and inhibited the expression of MMP9. Therefore, miR-218-5p may play a role as a tumor suppressor gene in gliomas. High expression of miR-218-5p can inhibit the dry maintenance of SHG-139s and its invasion ability.