Expressional Influence Of SiRNA-targeting Gene FMNL-2on Gene MMP-2in Primary Hepatocellular Carcinoma

Objective:1、To investigate the expression level of gene FMNL-2and MMP-2inhepatocellular carcinoma tissues, and to explore their relationship with clinicopat-hological features parameters of hepatocellular carcinoma;2、To preliminary explore the possible molecular mechanisms of FMNL-2andMMP-2in regulate the development, invasion and metastasis of hepatocellularcarcinoma cells.Methods:1、 Using immunohistochemistry MaxvisionTMmethod to detect the proteinexpression of FMNL-2and MMP-2in100cases of primary hepatocellular carcinomatissue and corresponding non-cancerous liver tissue.2、To detect the expression of FMNL-2and MMP-2in three strains of differentmetastatic potential of human hepatoma cell line by RT-PCR technique, screening thehepatocellular carcinoma cell lines with high expression of FMNL-2and MMP-2.3、 Using transient transfection of siRNA liposome technology to build thehepatoma cell line with low expression of FMNL-2.4、To detect the expression of MMP-2after interfering FMNL-2expression inhepatocellular carcinoma cell lines by RT-PCR technology.Results:1、 Immunohistochemistry test results: IOD of FMNL-2between the cancertissues and the adjacent tissues were90748.36±46276.91and39075.46±24763.12,IOD of MMP-2between the cancer tissues and the adjacent tissues were87969.46±27131.77and59517.72±20218.23(P<0.05); IOD of FMNL-2betweencirrhosis of the liver tissue and non-cirrhotic liver tissue were47471.96±40273.54and72123.41±45517.93（P<0.05）; IOD of MMP-2between HBsAg positive and negative group were77668.34±27682.79and64585.83±26173.62（P<0.05）, IOD ofMMP-2between metastasis and without metastasis group were63854.73±21730.42and77218.05±28930.01（P<0.05);The expressions of FMNL-2was correlated to theexpressions of MMP-2in the cancer tissues and the adjacent tissues (r=0.240，P=0.016).2、RT-PCR test results：FMNL-2mRNA expression levels in the MHCC-97H,HepG2and QGY-7703cell lines were（1.19±0.12）（,0.95±0.26）and（0.54±0.09）,MMP-2mRNAexpression levels in the three cell lines were（0.78±0.20）,（0.34±0.05）and （0.21±0.03）; In MHCC-97H hepatoma cells, the expression levels ofFMNL-2and MMP-2mRNA were significantly stronger than HepG2and QGY-7703hepatoma cells; After the downregulation of gene FMNL2by siRNA technology,FMNL-2mRNA expression levels in normal control group, negative control groupand siRNA-FMNL2group wer（e0.86±0.03）（,0.90±0.10）and（0.24±0.02）, MMP-2mRNA expression levels in normal control group, negative control group andsiRNA-FMNL2group were （1.44±0.04）,（1.17±0.05）and （0.31±0.01）;Conclusion:1、Compared with the adjacent noncancerous liver tissue, FMNL-2and MMP-2were highly expressed in the hepatocellular carcinoma tissue, both of them werenegatively correlated with each other;2、FMNL-2and MMP-2in human hepatocellular carcinoma cell line with highmetastatic potential were significantly higher than in low ones, suggesting that boththe two genes are closely related to the invasion and metastasis of hepatocellularcarcinoma.3、After the downregulation of gene FMNL2in hepatocellular carcinoma celllines with high metastatic potential by siRNA technology，MMP-2mRNAexpressionlevels were significantly reduced, suggesting that gene FMNL-2may be an upstreamregulator of gene MMP-2in hepatocellular carcinoma involved in the development ofliver cancer.