miR-1254 inhibits the proliferation and invasion of glioma cells by targeting CSF-1
10.3760/cma.j.cn371439-20200416-00082
- VernacularTitle:miR-1254通过靶向CSF-1抑制胶质瘤细胞的增殖和侵袭能力
- Author:
Liang CHEN
1
;
Jun QIN
;
Junrong LEI
;
Jun LIU
;
Lu WANG
Author Information
1. 湖北省十堰市太和医院（湖北医药学院附属医院）神经外科 442000
- From:
Journal of International Oncology
2020;47(10):577-584
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expressions of miR-1254 and its target gene colony-stimulating factor-1 (CSF-1) in glioma tissues and glioma cells, and the effects of miR-1254 and CSF-1 on the proliferation and invasion of glioma cells.Methods:The clinicopathological specimens and paracancerous tissues of 30 patients with glioma who underwent surgical treatment in Shiyan Taihe Hospital of Hubei Province from April 2017 to May 2019 were collected. Quantitative real-time fluorescent PCR (qRT-PCR) was used to detect the expression levels of miR-1254 and CSF-1 mRNA in glioma tissues, paracancerous tissues, U87 cells and human brain normal glial cells HEB. The glioma U87 cells were divided into blank control group, mimic NC group, miR-1254 mimic group, and siRNA NC group, CSF-1 siRNA group. The expression levels of miR-1254 and CSF-1 mRNA were detected by qRT-PCR. The protein expression levels of CSF-1 in each group were detected by Western blotting. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-1254 and CSF-1 gene. CCK-8 method and Transwell invasion experiment were used to detect the proliferation and invasion ability of cells.Results:The relative expression of miR-1254 mRNA in glioma tissues was 0.44±0.16, that in adjacent tissues was 1.15±0.28, and there was a statistically significant difference ( t=12.914, P<0.001). The relative expression of CSF-1 mRNA in glioma tissues was 1.96±0.27, that in adjacent tissues was 0.98±0.22, and there was a statistically significant difference ( t=14.970, P<0.001). The relative expression of miR-1254 mRNA in glioma cells U87 was 0.39±0.11, that in human brain normal glial cells HEB was 1.03±0.02, and there was a statistically significant difference ( t=10.113, P=0.008). The relative expression of CSF-1 mRNA in glioma cell U87 was 1.02±0.03, that in human brain normal glial cell HEB was 0.32±0.13, and there was a statistically significant difference ( t=9.037, P=0.009). The expression levels of CSF-1 mRNA and protein decreased with the increase of miR-1254 after transfection of miR-1254. The results of dual luciferase reporter gene assay showed that compared with the mimic NC group (1.04±0.12), the fluorescence activity of CSF-1-WT cells in the miR-1254 mimic group (0.31±0.02) was significantly reduced ( t=10.430, P<0.001). The difference in cell proliferation ability among the blank control group (0.71±0.01), mimic NC group (0.68±0.04) and miR-1254 mimic group (0.35±0.01) was statistically significant 48 h after transfection ( F=252.651, P<0.001). The difference was statistically significant between miR-1254 mimic group and blank control group ( P<0.001), and the difference was also statistically significant between miR-1254 mimic group and mimic NC group( P<0.001). The difference in cell proliferation ability among the blank control group (0.71±0.01), siRNA NC group (0.68±0.04) and CSF-1 siRNA group (0.25±0.01) was statistically significant ( F=320.309, P<0.001). The difference was statistically significant between CSF-1 siRNA group and blank control group ( P<0.001), and the difference was also statistically significant between CSF-1 siRNA group and siRNA NC group ( P<0.001). Invasion experiments showed that the difference of transmembrane cells number among the blank control group (365±27), mimic NC group (388±24) and miR-1254 mimic group (83±15) was statistically significant ( F=173.915, P<0.001). The blank control group (365±27), siRNA NC group (404±32) and CSF-1 siRNA group (87±14) had statistically significant difference in the number of transmembrane cells ( F=141.294, P<0.001). Conclusion:The expressions of miR-1254 in glioma tissues and glioma cells U87 are significantly decreased, and the expressions of CSF-1 in glioma tissues and glioma cells U87 are significantly increased. Overexpression of miR-1254 may inhibit the proliferation and invasion of glioma U87 cells by reducing the expression of CSF-1 targetedly.
