The Correlation Between Synergy Of UPAR（D1D2） And Integrin α₅β₁with Occurrence Of Liver Cancer

Objectives:To choose the best method of liver cell culture in order to provide abundant source of specimens and detect the the expression ofuPAR (D1D2)mRNA in normal liver cells, HCC arcinomaadjacent cells and HCC cells. At the same time,to detect the expression of uPAR (D1D2)and integrinα5β1in normal liver tissue, HCCadjacent tissue and HCC tissues, in Guangxi population and explore the characteristics and relevance of them in the process of HCC occurrence.Methods:Comparing tissue block adherent method, liver cell grindingmethod and pancreatic enzyme digestion used in normal liver tissue to choosethe best culture method.Used the best culture method to culture normal livercells, HCC arcinomaadjacent cells,HCC cells and detect the expression ofuPAR (D1D2)mRNA in them.At the same time,the mRNA were extracted fromthe60cases of hepatocellular carcinoma,60cases of adjacent tissues and25cases of normal liver tissues.RT-PCR was used to detect the mRNA expressionof uPAR(D1D2) and integrinα5β1in three tissues.At the same time,we used insitu hybridization to detect the expression sites of uPAR(D1D2) and integrinα5β1 mRNA in60cases of hepatocellular carcinoma, the60cases of carcinoma and25cases of normal liver tissue,and the relative content of them.Results:1.Comparing tissue block adherent method, liver cell grindingmethod and pancreatic enzyme digestion used in normal livertissue,the liver tissue adherent method was economic,simple and thebest method for liver cell culture.2.Used RT-PCR to detect the expression of uPAR (D1D2)mRNA innormal liver cells, HCC arcinomaadjacent cells, HCC cells.Wefound that the brightness of the cells obviously increase from normal liver cells, para-carcinoma cells and liver cancer cells. Comparison of the cell grey values of three groups, P<0.05,it showed statistically significant difference.3.We found that the expression of uPAR(D1D2) and integrinα5β1obviously increase from normal liver, para-carcinoma tissue andliver cancer tissue in RT-PCR. Comparison of the greyvalues of threegroups,P<0.05, it showed signifitissue cant difference.4. In situ hybridization result showed that the express of uPAR(D1D2) and integrinα5β1in all the cytoplasm the positive rate of both in HCC tissue were significantly higher than para-carcinomatissue and normal liver tissue.The expression levelsand were determined comprehensively by positive cells rate and staining intensity scores.(P<0.05)5. By spearman rank correlation analysis the expression level of uPAR (D1D2) and integrinα5β1in HCC, and found a positive correlation between them (=0.257, P <0.05; rs2=0.261, P <0.05). Itconsidered that there is a synergistic effect between uPAR (D1D2) and integrinα5β1in the development of liver cancer.Conclusion:1.The liver tissue adherent method was economic,simple a nd thebest method for liver cell culture. It can culture and separate out liver cells with high density, high purity and high activity.2. The brightness of uPAR (D1D2)mRNA was obviously increased from normal liver cells, para-carcinoma cells and liver cancer cells.It prompts that uPAR (D1D2) was associated with malignant transformation of liver cells.3.The expression of uPAR (D1D2) and integrinα5β1were increased significantly from normal liver tissue, para-carcinoma tissue and liver cancer tissue,prompted that the synergy of uPAR(D1D2) and integrinα5β1interactionwillcause liver cells abnormal clonal hyperplasia and malignant transformation in Hepatic Cell.