Relationship Between Expression Of S100P And Hepatocellular Carcinoma With Vascular Invasion

Objective : This study mainly explored the relationship between S100 P expression and hepatocellular carcinoma with vascular tumor thrombus.Methods : The specimens were selected from HCC patients who underwent surgical resection in hepatobiliary and pancreatic spleen surgery and confirmed by pathology at the Affiliated Tumor Hospital of Guangxi Medical University from January 2015 to December 2016.Patients were divided into five groups: HCC with PVTT?PVTT?HCC with MVI(II)?HCC with MVI(I)?HCC with MVI(I0).RT-QPCR and Western Blot were respectively used to detect the expression level of S100 P mRNA and protein in tissues.Kaplan-Meier method combining log-rank test were used to estimate the recurrence-free survival time between groups of S100 P high expression and low expression.Two different types of S100P-shRNA lentiviral vector were constructed to silence the expression of S100 P respectively in HCC cell line SMMC-7721 and LM3.Cellular function experiments were performed after 48 hours of successful transfection to respectively detect the ability of proliferation ? migration ?invasion?apoptosis in hepatocarcinomatous cells between S100 PshRNA group and negative-control group.Western blot was used to detect the changes of protein markers including E-cadherin,EPCAM,N-cadherin,Twist1 and Vimentin in the EMT(epithelial-mesenchymal transition)pathway in terms of silence of S100 P.Results : 1.RT-qPCR and Western blot showed that the expression of S100P(mRNA and protein)was highest in group of HCC with PVTT and PVTT,secondly in group of HCC with MVI and lowest in group of adjacent-tumor tissue(P<0.05).S100P(mRNA and protein)in group of MVI(II)was highest among groups of MVI(II)?MVI(I)and MVI(I0).Compared with low expression of S100P(mRNA and protein),high expression of S100P(mRNA and protein)showed significant shorter RFS(P<0.05).2.The results of CCK8 assay for cell proliferation assay showed that the OD value of S100 P gene silencing group was significantly decreased compared with the untreated cell line group on the fourth day of transfection,and the difference was statistically significant(P<0.05).3.Cell scratch test showed that compared with the untreated cell line group,the cell migration rate of S100 Psh group was significantly decreased at 48 h and72h,the difference was statistically significant(P<0.05).4.Transwell invasion experiment: Compared with the untreated cell line group,the number of transmembrane cells in the S100 Psh group was significantly decreased,and the difference was statistically significant.(P<0.05).5.Flow cytometry was used to detect apoptosis.After S100 P silencing,the apoptosis rate of S100 Psh group was significantly increased,and the difference was statistically significant(P<0.05).6.After S100 P gene silencing,the expression levels of E-cadherin,N-cadherin and EPCAM in the ETM pathway were significantly increased,and the difference was statistically significant(P<0.05).Conclusion : Overexpression of S100 P was significantly associated with hepatocellular carcinoma with portal vein invasion and metastasis.S100 P may played an important role in EMT pathway.