Effect Of MTIF2 On Apoptosis,Proliferation And Migration Of Hepatocellular Carcinoma Cells

Objective: The correlation between mitochondrial translation initiation factor 2(MTIF2)and hepatocellular carcinoma(HCC)cell’s proliferation,migration,apoptosis and 5-fluorouracil was investigated by compared the changed in migration,proliferation,apoptosis under 5-fluorouracil treated in HCC cells by the MTIF2 down-regulated.Methods: First,western blot was applied to detect MTIF2 gene expression in four types of cells including human normal hepatocytes L02,human hepatoma cells HepG2,Bel-7402,and Huh7,and we found that the MTIF2 gene was highly expressed in HepG2 cells.So this experiment used HepG2 cells as the study object.HepG2 cells were divided into two groups for the experiment.One group was classified as an experimental group(HepG2-sh MTIF2)by constructing an sh RNA interference fragment targeting MTIF2 to down-regulate MTIF2 in HCC cells.The other group was not specially treated and was categorized as the control group(HepG2-NC).The changes in apoptosis rate,proliferation,and migration between the two groups of HCC cells were evaluated.Among them,the proliferation of HCC cells was analyzed by CCK8 and Ed U assays;cell migration and invasion were performed by wound healing and transwell assays;and then,we performed a preliminary investigation of the relationship between MTIF2 and 5-FU chemotherapeutic agents for HCC by comparing the changes in viability and apoptosis rate of HepG2 cells subjected to 5-FU before and after MTIF2 interference.The apoptosis rate of HCC cells was detected by annexin V staining.Finally,the potential impact of MTIF2 on the prognosis of HCC was assessed by querying the the Cancer Genome Atlas(TCGA)database for univariate and multifactorial Cox regression analysis of MTIF2 expression in patients with HCC.Results:Among the four hepatocyte cell lines appealed,western blot found higher expression of MTIF2 in HepG2,and subsequently.we applied lentiviral RNA interference technology for human MTIF2 sh RNA to design two si RNA knockdown targets(sh RNA#1,sh RNA#2).In the effects of sh RNA,MTIF2-sh RNA#2 showed a better interference effect,so it was selected for subsequent experiments.An assessment of the effect of MTIF2 on the proliferation of cancer cells,CCK8 revealed that the proliferation of HCC was significantly inhibited in the sh MTIF2 group compared with the NC group after MTIF2 down-regulated(P < 0.05).And this finding was further confirmed by the Ed U cell proliferation assay,in which MTIF2 down-regulated significantly affected the proliferation of HCC cells(P < 0.001).In the correlation analysis between MTIF2 and HCC cell migration and invasion,the results of wound healing and transwell assay suggested that the scratch area of NC group was significantly reduced and the number of cell migration was135±12.36 and 69±8.41 for HepG2-NC and HepG2-sh MTIF2,respectively,at 24 h reculture after cell surface manufacturing trauma(p<0.001).The number of HepG2-sh MTIF2 group HCC cells crossed Matrigel gel in 99.80±15.82,201,which was significantly lower than that of MTIF2 control NC group HCC cells crossed Matrigel gel in203.6±17.17(P<0.001),which indicated that MTIF2 down-regulated could considerably inhibit the migratory and invasive ability of HCC.The correlation analysis between MTIF2 and HCC cell apoptosis revealed that MTIF2 down-regulated can significantly increased the apoptosis rate of cells in the sh MTIF2 group(22±2.16)compared to the NC group(7.5±2.38)(P<0.001).CCK8 and apoptosis assays under 5-fluorouracil treatment showed that down-regulated MTIF2 increased 5-FU’s therapy to HCC and further inhibited cell activity and its proliferative capacity.And compared with the NC-5-FU group(21.80±4.47),the apoptosis rate of sh RNA-5-FU group transfected with MTIF2 targeting sh RNA after24 h treatment with 5-FU was 64.00±15.91(P<0.001),which indicated that MTIF2 down-regulated could enhance the pro-apoptotic effect of5-FU on HCC.Finally,regression analyses of univariate and multifactorial Cox showed that MTIF2 expression was an independent prognostic risk factor.Conclusion Firstly,MTIF2 may be involved in the negative regulation of apoptosis and the positive regulation of proliferation,migration,and invasion of hepatocellular carcinoma cells;secondly,down-regulated MTIF2 can significantly enhance the cytotoxicity and apoptosis effect of 5-fluorouracil in HCC cells.Thirdly,MTIF2 is closely related to the clinical progression of HCC,and the high expression of MTIF2 may be negatively correlated with tumor prognosis,the specific mechanism of which deserves further in-depth study.