Experimental Study
The mechnism and effect of microRNA-518b on cell proliferation, apoptosis and invasion in hepatocellular carcinoma cell
Rixin Zhang, Lei Liu, Chaowen Xiao, Kai Li, Zhi Zheng, Ling Zhu
Published 2019-07-08
Cite as Chin J Exp Surg, 2019, 36(7): 1208-1210. DOI: 10.3760/cma.j.issn.1001-9030.2019.07.014
Abstract
ObjectiveTo investigate the effect of microRNA (miRNA, miR)-518b on proliferation, apoptosis and invasion of hepatocellular carcinoma cells.
MethodsThe miR-518b expression was measured by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) in hepatocellular carcinoma and normal hepatocyte lines. HepG2 cell line was divided into miR-518b over-expressed group (miR-518b group), negative control group (miR-Con group) and blank control group (Blank group). Methyl thiazol tetrazolium (MTT) assay, flow cytometry and Transwell assay were used to measure cell proliferation, apoptosis and invasion. The activity of cysteinyl aspartate-specific protease (Caspase)-3, Caspase-8 and Caspase-9 was determined by spectrophotometry, and the expression of Rap1b protein was determined by Western blotting. T-test was used for comparison of measurement data.
ResultsThe expression of miR-518b in hepatocellular carcinoma cell lines was lower than that in normal hepatocellular cell line (P< 0.05). The A490 nm of miR-518 group was lower than miR-Con group at 72, 96 h after transfection (0.66±0.07 vs. 0.94±0.08, t=3.291, P<0.05) and (0.86±0.07 vs. 1.47±0.10, t=3.943, P<0.01). The apoptotic rate in the miR-518b group was higher than that in miR-Con group and Blank group [(21.4±3.2)% vs. (7.5±1.2)% vs. (6.8±1.1)%, t=13.287, P<0.05]. The relative activity of Caspase-3 and Caspase-9 in miR-518b group was highest in three groups (2.10±0.20 vs. 1.02±0.04 vs. 1.04±0.05, t=16.287, P<0.05), (2.20±0.20 vs. 1.03±0.03 vs. 1.04±0.03, t=16.298, P<0.05). There was no significant difference in the relative activity of Caspase-8 between miR-518b group, miR-Con group and Blank group (t=8.288, P>0.05). The number of invasive cells in miR-518b group was less than that in miR-Con group and Blank group (74±10 vs. 151±15 vs. 153±12, t=17.276, P<0.05). The expression of Rap1b protein in miR-518b group was lowest in three groups (0.38 ± 0.03 vs. 1.03±0.06 vs. 1.05±0.04, t=19.642, P<0.05).
ConclusionThe down-regulation of the expression of miR-518b was founded in hepatocellular carcinoma cell lines. Over-expression of miR-518b inhibits cell proliferation and invasion, and induce apoptosis. The mechanism may be related to the up-regulation of Caspase-3, Caspase-9 expression and the down-regulation of Rap1b expression.
Key words:
MicroRNA-518b; Hepatocellular carcinoma; Proliferation; Apoptosis; Invasion
Contributor Information
Rixin Zhang
Department of Hepatobiliary &
Pancreatic Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
Lei Liu
Department of Gastrointestinal Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
Chaowen Xiao
Department of Hepatobiliary &
Pancreatic Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
Kai Li
Department of Hepatobiliary &
Pancreatic Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
Zhi Zheng
Department of Hepatobiliary &
Pancreatic Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
Ling Zhu
Department of Hepatobiliary &
Pancreatic Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China
