IFITM3 Expression And Its Regulatory Action Over MMP9 In Hepatocellular Carcinoma

Objective:1.To detect the expression of interferon-induced transmembrane protein 3(IFITM3)in hepatocellular carcinoma(HCC),and to evaluate the correlation between expression level of the protein and clinical pathology features.2.To investigate the changes in invasion and migration ability of hepatocellular carcinoma after IFITM3 regulating matrix metalloproteinase 9(MMP9).Methods:1.The protein expression of IFITM3 along with matrix metallopeptidase 9(MMP9)in 60 specimens of HCC and paired cancer adjacent tissues were determined by immunohistochemical staining and Western blot analysis,and to evaluate the correlation between expression level of the IFITM3 protein and clinical pathology features by statistical analysis.2.IFITM3 small interfering RNA fragments(psilencer3.1-sh IFITM3)were constructed and transfected into HCC Hep G2 cells by Liposome-mediated,and choose the most effective interfering RNA fragments by Western blot detecting the changes in IFITM3 protein of Hep G2 cells after transfection.3.The experiment was divided into the Control cells group,Scrambled si RNA group and IFITM3 si RNA group,and after transfection,the changes in m RNA and protein expressions of IFITM3 and MMP-9,the proliferative capacity,as well as the invasion and migration ability of the Hep G2 cells were examined by q RT-PCR,Western blot,CCK8 assay,and Transwell and scratch wound-healing assay,respectively.Results:1.Both positive expression rate and expression level of IFITM3 and MMP-9 protein expressions in HCC tissue were signific antly increased compared with cancer adjacent tissue(positive expression rate: 81.67% vs.13.33%;88.33% vs.8.33%,all P<0.05),and it has a positive correction between IFITM3 and MMP-9 protein expressions in HCC tissue with correlation coefficient(r)was 0.476(P<0.01).2.The high expression level of IFITM3 in HCC tissue has a positive correction between large tumor size,tumor tissue encapsulation,microvascular invasion,high rates of intrahepatic metastasis,high TNM stage,and poor differentiation(P<0.05),and it has no positive correction between age,gender,cirrhosis,AFP,and other clinical pathology features(P>0.05)3.hs-IFITM3-si-2 was the most effective IFITM3 small interfering RNA fragments compared with the Control cells group and Scrambled si RNA group.4.Both IFITM3 and MMP-9 expression in Hep G2 cells after psilencer3.1-sh IFITM3 transfection were decreased at either m RNA or protein level,proliferation rate and the number of invaded cells were reduced,and wound closure was significantly delayed.The differences in all quantitative parameters had statistical significance(all P<0.05).It confirmed that IFITM3 regulated cell invasion and migration through regulating MMP9 expression.Conclusion:1.IFITM3 expression is increased in HCC and has a positive correction with large tumor size,tumor tissue encapsulation,microvascular invasion,high rates of intrahepatic metastasis,high TNM stage,and poor differentiation(P<0.05).2.The high expressed IFITM3 may promote the proliferation,invasion and migration of HCC cells through regulating MMP-9 expression,and IFITM3 may serve as a potential target of gene therapy for HCC in the future.