Mechanism of micro RNA-1182 overexpression in inhibiting malignant phenotype of glioma cells
10.3760/cma.j.cn115354-20210519-00319
- VernacularTitle:过表达miR-1182抑制脑胶质瘤细胞恶性表型的机制研究
- Author:
Zhihao ZHAI
1
;
Bin LUO
;
Xiaowen LIAN
;
Jianliang CHEN
;
Yuanqiang ZHONG
;
Hengxing YOU
Author Information
1. 中山大学附属第八医院神经外科，深圳 518000
- Keywords:
Glioma;
Micro RNA-1182;
Malignant phenotype;
Cell cycle-related protein;
Phosphatidylinositol 3 kinase/protein kinase B pathway;
Epithelial-mesenchymal tra
- From:
Chinese Journal of Neuromedicine
2021;20(10):973-980
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the micro RNA (miR)-1182 expression in glioma, and explore the regulation role and mechanism of miR-1182 overexpression in malignant phenotype of glioma cells.Methods:(1) The data of miR-1182 expressions of 198 glioma samples and survival of these glioma patients were downloaded from the official website of Chinese Glioma Genome Atlas(CGGA), and the differences of miR-1182 expression levels among glioma tissues of different pathologic types and different WHO grades were compared. Kaplan-Meier survival curve was used to analyze the relation between miR-1182 expression level and patient survival. (2) Human glioma cell lines A172, LN229, T98G, U87, and U251, and human normal astrocyte cell line NHA were routinely cultured in vitro, and the miR-1182 expression levels in each group were detected by real-time quantitative PCR (qPCR). (3) U87 and U251 cells were divided into miR-1182 transfection group and negative control group; the miR-1182 mimics and miR-1182 negative control sequence were transfected, respectively. After 48 h of transfection, 5-ethynyl-2'-deoxyuridine (EdU) staining was used to detect the cell proliferation ability, flow cytometry was used to detect the cell apoptosis, Transwell assay was used to detect the cell migration ability, and Western blotting was used to detect the expression levels of cyclin (C-myC, C-Jun, CCND1, and P21), phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt), and epithelial-mesenchymal transformation (EMT) pathway related proteins (N-cadherin, β-catenin, and vimentin). Results:(1) The miR-1182 expressions in glioma tissues of WHO grading III and IV were significantly lower as compared with those in glioma tissues of WHO grading II ( P<0.05). The median survival time in patients from the low miR-1182 expression group ([701.00±11.14] d) was significantly shorter than that in the high miR-1182 expression group ([1812.00±23.21] d, P<0.05). (2) As compared with that in NHA cell group, the miR-1182 expression levels in A172, LN229, T98G, U87 and U251 cell groups were significantly decreased ( P<0.05), and the decrease was most significant in U87 and U251 cell groups. (3) As compared with the negative control group, the U87 and U251 cells in miR-1182 transfection group had significantly weaker proliferation ability, significantly higher apoptosis rate, significantly decreased number of transmembrane cells, significantly decreased protein expression levels of C-MyC, C-Jun and CCND1, significantly increased P21 protein expression level, significantly decreased expression levels of PI3K, phosphorylated (p)-PI3K, Akt and p-Akt, and significantly decreased expression levels of N-cadherin, β-catenin and vimentin ( P<0.05). Conclusions:Glioma patients with low miR-1182 expression have poor prognosis. Low miR-1182 expression is noted in glioma cells. Overexpression of miR-1182 can inhibit the malignant phenotype of glioma cells, which may be related to cell cycle-related proteins, PI3K/Akt, and EMT pathway ralated proteins.
