The effects of microRNA-7 on proliferation and invasion of hepatocellular carcinoma HepG2 cells
10.3760/cma.j.issn.0253-3766.2018.06.002
- VernacularTitle: 微小RNA－7对肝癌HepG2细胞增殖和侵袭能力的影响
- Author:
Andong QIN
1
;
Xingxiang LIU
2
;
Jing LI
3
;
Juan LIU
2
;
Yusong LI
2
Author Information
1. Research Institute of Liver Disease, the Fourth People′s Hospital of Huai′an, Huai′an 223002, China
2. Clinical Laboratory, the Fourth People′s Hospital of Huai′an, Huai′an 223002, China
3. Department of Liver Disease, the Fourth People′s Hospital of Huai′an, Huai′an 223002, China
- Publication Type:Journal Article
- Keywords:
Hepatocellular carcinoma;
HepG2 cells;
MicroRNA-7;
Raf1;
Proliferation
- From:
Chinese Journal of Oncology
2018;40(6):406-411
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro.
Methods:The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3′UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis.
Results:The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05).
Conclusions:The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.
