Mechanisms Of Sodium Aescinate Inhibiting Proliferation Of Hepatoma Cells Through CARMA3/NF-?B Signaling Pathway

Purpose: Primary hepatocellular carcinoma(HCC)is a very malignant tumor in the world.It has a high incidence in China and accounts for half of the worldwide incidence.Because of its high degree of malignancy,it is not sensitive to chemoradiotherapy and chemotherapy and is found to be late in time.Poor prognosis,and high recurrence rate,and other factors seriously threaten the health of our people.For the treatment of early stage primary HCC cancer is still mainly the surgical treatment,but for patients with advanced liver resection is not optimistic,with the molecular biology and tumor gene related research continues to deepen,the new molecular targeted drug research and development,hoping to become a new breakthrough in the treatment of liver cancer.Related reports reported a 5-year recurrence rate of liver cancer nearly 70%,so the recurrence and metastasis of liver cancer problems seriously shortened the survival of patients and greatly reduce the quality of life of patients.Therefore,it is very important to study the molecular mechanism of proliferation and metastasis of HCC and so on.This research mainly studies the expression of CARMA3 in human hepatocellular carcinoma and the potential mechanism of sodium aescinate in inhibiting the proliferation of human hepatoma cells and expecte to find effective related protein markers and provide a scientific basis for clinical application of sodium aescinate antitumor.Methods: 100 cases of pathologically confirmed primary hepatocellular carcinoma were enrolled in this study.Tissue microarray immunohistochemistry was used to detect the expression of CARMA3 and NF-?B in 100 cases of hepatocellular carcinoma and its correlation with clinicopathological prognosis.The CARMA3 gene was overexpressed and knocked down by using plasmid transfection in vitro.Then the proliferation,apoptosis and clonogenic capacity of hepatoma cells were detected after transfection.MTT assay was used to detect the proliferation of hepatocellular carcinoma cells treated by sodium aescinate and determine the optimum drug concentration;flow cytometry was used to detect HCC cells apoptosis treated by sodium aescinate IC50 concentration;Western blot and immunofluorescence assay were used to detect expression of CARMA3 / NF-?B in Hep G2 and Hep3 B Cell lines treated with sodium aescinate.Results: Tissue microarray analysis of 100 patients showed that CARMA3 was over-expressed in HCC tissues compared with paired adjacent liver tissues and was significantly prognostic survival relevant.In vitro,knockdown of CARMA3 inhibited the proliferation,colony formation,and induced the early-stage apoptosis of HCC cell lines.The over-expression of CARMA3 in the same cell lines showed the adverse results.Sodium aescinate is reported in this study to inhibit the expression of CARMA3 and its downstream kinase NF-?B for the first time.Moreover,Sodium aescinate inhibited the growth of HCC cell lines without toxic effects to normal hepatocytes.Over-expression of CARMA3 by plasmid transfection partly redressed the anti-tumor effects of Sodium aescinate.Conclusions: Our data suggest that CARMA3/NF-?B is overexpressed in HCC tissues and correlates with tumor prognosis,which may be an effective target protein in the treatment of liver cancer.Meanwhile,sodium aescinate can effectively inhibit the activation of CARMA3/NF-?B in HCC to inhibit the proliferation of primary hepatocellular carcinoma,which may be a new molecularly targeted drug for the treatment of liver cancer.