The Expression And Significance Of S100A4,MMP-9 In Hepatic Carcinoma Cell Lines And Hepatocellular Carcinoma Tissue

S100 protein is a kind of calcium-binding regulatory proteins,and it was first found by moore from the brain of moo-cow.As its biochemical property of remaining soluble after precipitation with 100%ammonium sulfate,the proteins are called S100.S100A4 protein,a member of S100 family,which derives from mammal epithelial cells with highmetastatic potential and some kinds of carcinoma cells,exhibits dual,intracellular and extracellular biological functions.People conclude that the S100A4 protein plays an important role in the progress of tumor invasion and metastasis.In this research,immunohistochemical method is used to detect the expression of S100A4 and MMP-9 at the protein level in hepatocellular carcinoma tissue and different human hepatic carcinoma cell lines,and we analyze the relationship between their expression and biological behavior of HCC,approach the possibility of using S100A4,MMP-9 as molecular tumor marker to appraise the latent potential of invasion and metastasis,and thus provide pathological foundation for research and clinical therapy of HCC.Methods In this research culture MHCC97H,MHCC97L,Hep3B,HepG2 human hepatic carcinoma cell lines and L02 human normal hepatic cell lines,After cells growing on cover glass,do immunoeellularchemical pigmentation.And 36 paraffin-embedded cancer biopsies of patients with HCC through pathology diagnosis in Shandong Provincal Hospital from January 2006 to November 2007.Before the operation, did not give preoperative adjunctive therapy,like transcatheter arterial chemoembolization (TACE)and chemical therapy.We also collected 14 fresh specimens of patients.The fresh specimens include the tumor tissue,the adjacent nontumorous liver tissue(the distance to tumor＞3cm),tumor and adjacent tissue,was stored in -80℃.As controls,8 health adult liver specimens were wedge biopsies during surgeries from patients with other abdominal lesions or hepatic hemangioma.All the specimens are detected by immunohistochemistry.The data have been statistically analyzed with Chi square test, Pearson correlation and Spearman rank correlation using SPSS 13.0 for windows.Result(1)Positive expression of protein S100A4 is not observed in human normal hepatic cell line L02,human hepatic carcinoma cell line Hep3B without metastasis potential.The positive expression of protein S100A4 can be observed in human hepatic carcinoma cell line HepG2,MHCC97L and MHCC97H with metastasis potential.The expression of the protein in high metastasis cell line MHCC97H is higher than that in low metastasis cell line MHCC97L and Hep G2;(2)Positive expression of protein MMP-9 is not observed in human normal hepatic cell line L02.In Hep3B,he positive expression of protein S100A4 can be observed,but the expression intensity is lower than in human normal hepatic cell line with metastasis potential;(3)Statistical analysis shows that protein S100A4 and MMP-9 expressions are positively related in different cell lines;(4)Positive expression of protein S100A4 is not observed in human normal hepatic tissue.While in HCC tissue,elevated levels of S100A4 are detected(the positive rate is 64.0%),and the elevated levels of S100A4 is high in HCC metastasis tissue(the positive rate is 77.78%);(5)Positive expression of protein MMP-9 is observed in the 8 normal liver tissues and different HCC tissue.The positive expression rate of MMP-9 in HCC metastasis tissue is 88.89%(16/18),more high than 62.5%(20/32)of HCC tissue without metastasis.6.Statistical analysis shows that protein S100A4 and MMP-9 expressions are positively related in HCC.Conclusions(1)In human hepatic carcinoma cell lines HepG2,MHCC97L and MHCC97H,there are positive protein S100A4 and MMP-9 expression,and the expression intensity is increased as the ability of metastasis potential.It can be inferred that the two proteins plays an important role in the biological behaviors of HCC cells;(2)In metastasis HCC tissue,the expression rate of protein S100A4 and MMP-9 is higer than that in HCC tissues.There are significant differences about the expression rate between the group with metastasis and the group with not.The high expressions of S100A4 cooperate with MMP-9 may take part in HCC invasion and metastasis;(3)For human hepatic carcinoma cell lines and HCC,there are positive correlation between protein S100A4 and MMP-9 expressions.S100A4 protein may participate in mediating the expression of MMP-9,and accelerate the degradation of extracellular matrix and basal membrane which enhance the HCC cells invasion and metastasis;(4)It can be used as molecular tumor markers to appraisal the latent energy of invasion and metastasis of tumor and thus provide pathological foundation for research and clinical therapy of HCC to detect the expressions of protein S100A4 and MMP-9 in the use of immunohistochemical EnVision add Dual Link System Peroxidase.