Studies On The Correlation Between SERPINB1and Hepatocellular Carcinoma

ObjectiveTo investigate the expression of SERPINB1in the hepatocellular carcinoma (HCC),and analyze the correlation of SERPINB1abnormal expression and the occurrence anddevelopment of HCC. To futher reveal the molecular pathogenesis of HCC, and providethe new target for therapy of HCC.Methods1. The expression of SERPINB1in8cases of fresh frozen specimens from HCC andthe adjacent liver tissue were detected by Western blot. Immunohistochemical to detect theexpression of SERPINB1in67samples of HCC tissue and adjacent non-tumor tissue. Toanalyze the connection between SERPINB1and clinicopathologic datas of HCC patients,such as age、gender、histological grade、metastasis、vein invasion、the serum of AFP andso on.67cases of HCC patients were followed up, survival curve was calculated using theKaplan-Meier method, and the prognosis was analyzed with the univariate and multivariateCox proportional hazards regression analysis.2. The expression of SERPINB1was detected in normalhepatocyte cell line LO2and three human HCC cell lines: HuH7, HepG2, BEL-7404.HuH7and BEL-7404cells were transfected with SERPINB1-targeting siRNA. The woundhealing and the matrigel invasion assay detected the migration and invasion of HuH7andBEL-7404cells. CCK8assay determine the biologic effects of the decreased expression ofSERPINB1, the growth rate of the HuH7and BEL-7404cells. Western blot was used todetect the expression levels of active form of MMP2in HuH7and BEL-7404cellstransfected with SERPINB1-targeting siRNAResults1. Immunohistochemistry and western blot analysis showed that SERPINB1expression in HCC were lower than that in non-tumor liver tissue. SERPINB1expressionwas correlated with metastasis(P=0.000) and invasion (P=0.006). The Kaplan-Meier survival curves showed that low SERPINB1expression related to a poor survival withstatistical significance (p=0.001). Unvaried analysis showed that SERPINB1expressionwas associated with poor prognosis of HCC (p=0.002). Multivariate Cox proportionalhazards regression analysis indicated that SERPINB1expression was an independentprognostic marker for HCC (p=0.023).2. Western blot was used to detect the expression of SERPINB1in LO2, HuH7,HepG2and BEL-7404. The level of endogenous SERPINB1in the three HCC cell lineswas lower thanthat in the normal hepatocyte cell line LO2. After SERPINB1-targeting siRNA wastransfected into HuH7and BEL-7404cells which have higher expression of SERPINB1,we found that SERPINB1inhibits the migration and invasion in HuH7and BEL-7404cells，while cell proliferation was not affected. we also found that knockdown ofSERPINB1significantly increased the level of the active MMP2.Conclusions1. The expression of SERPINB1was downregulated significantly in HCC.SERPINB1served as an independent prognostic marker for breast cancer. These datasrevealed that SERPINB1may play a role in the oncogenesis and development of HCC.2. During the migration and invasion process of HuH7and BEL-7404cells, theregulation of MMPs by SERPINB1may played a role in it.