Tianjin Med J ›› 2018, Vol. 46 ›› Issue (11): 1151-1154.doi: 10.11958/20180721

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The effect of lentivirus-mediated overexpression of microRNA-155 on hepatoma cell HepG2 proliferation

NIU Lian-jie1 , ZHANG Ya-min2△, WU Zhao-guo1   

  1. 1 The First Central Clinical College of Tianjin Medical University, Tianjin 300070, China; 2 Department of Hepatobiliary Surgery, Tianjin First Central Hospital
  • Received:2018-05-07 Revised:2018-09-10 Published:2018-11-15 Online:2018-11-20
  • Contact: lianjie NIU E-mail:niulianjie910107@163.com

Abstract: Abstract:Objective To investigate the effect of up-regulation of microRNA-155 (miR-155) on the proliferation of hepatoma cell line HepG2 by constructing lentivirus (LV) overexpressing miRNA-155. Methods The specific miR-155 overexpressing fragment was synthesized into the target sequence and cloned into the lentiviral vector pGCSHL-GFP. The recombinant miR-155-hRNA-LV and the negative control empty viral vector were transfected into HepG2 cells to establish stable overexpression miR-155 cell line. The miR-155 cells were divided into three groups including the overexpression group (H group), the negative control group (negative control, NC group) and the normal group (normal, N group). Real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-155 and PTEN in three groups. Western blot assay was used to analyze relative expressions of PTEN, CyclinD1 and CyclinA1+A2 proteins in three groups. Cell proliferation-toxicity test (CCK-8) was used to detect cell proliferation. Results The expression of miR-155 was significantly higher in H group than that in NC group and N group. The expression of target gene PTEN was lower in H group than that in NC group and N group (both P<0.05). The expression of PTEN protein was significantly lower in H group than that in NC group and N group, while the expressions of CyclinD1 and CyclingA1+A2 protein were significantly higher in H group compared with that of NC group and N group (both P<0.05). CCK-8 results showed that the photometric values of the three groups began to differ at 12 h, and the luminosity values were significantly greater in H group than those of NC group and N group from 24 h to 72 h, but the difference was not statistically significant between NC group and N group. Conclusion Overexpression of miR-155 can promote the proliferation of HepG2 cells.

Key words: liver neoplasms, hepatocytes, microRNAs, lentivirus infections, microRNA-155