Studies On The Anti-hepatocellular Carcinoma And Immunomodulatory Effects Of Calf Spleen Extractive Injection

Malignant neoplasms are one of the three major killers of human mortality,threatening human health.Hepatocellular carcinoma(HCC)is one of the most common cancers in the world,which killing 662,000 people worldwide annually,and wherein the cancer patients,about half of them are Chinese.In addition,the treatment of HCC is often accompanied by the poor prognosis,relapse and transfer.At present,the treatment of HCC mainly incudes chemotherapy,radiation therapy,surgery,gene therapy,percutaneous treatment and liver transplantation,but these methods are not only expensive but also cause serious side effects.As a multicomponent biochemical extract,Calf Spleen Extractive Injection(CSEI)has many targets and low toxicity.However,there is no systematic study on its anti-tumor activity and its related mechanism.In this paper,the immunomodulatory effect and anti-hepatocarcinoma activity of CSEI were studied.The main contents of this paper include the following aspects:1.Study on anti-hepatocarcinoma activity of CSEI and its mechanism in vitroHepatocellular carcinoma cell lines HepG2 and SMMC-7721 were used to investigate the proapoptotic effect of CSEI.At the same time,the mechanism of CSEI in promoting hepatoma cell apoptosis was investigated by biochemical analysis.The results showed that CSEI could inhibit the growth of hepatocellular carcinoma,breast cancer and lung cancer cell lines in vitro.The inhibitory effect of CSEI on hepatoma cell line SMMC-7721 and HepG2 was most obvious,and it could increase ROS production,changes the mitochondrial membrane potential,activate caspase cascade reaction.In addition,the expression of PARP,cleaved-PARP,Bcl2,P38,phospho-P38,JNK,phospho-JNK1,ERK1/2,phospho-ERK1/2,cleaved-caspase 3 cleaved-caspas 8 and cleaved-caspase 9 were detected by Western Blot.The expressions of oxidative stress-related proteins Nrf2 and Catalase were significantly decreased in hepatocellular carcinoma cells.In summary,CSEI inhibits the proliferation of hepatocellular carcinoma cell through the ROS-related mitochondrial pathway.2.Study on anti-hepatocarcinoma activity of CSEI and its mechanism in vivoIn order to further determine the anti-hepatocarcinogenesis effect of CSEI,the HepG2-xenografted tumor mice model was established.The body weights and tumor volume were recorded in the other day after CSEI treatment within 14 days.After the treatment,the expressions of anti-tumor and oxidative stress-related proteins were analyzed by Western blot.The experimental results showed that the CSEI significantly decreased the tumor volume of nude mice in HepG2-xenografted tumor mice model and had no effect on the body weights of mice.Similarly,Western blot results were consistent with the in vitro experiment.These results indicated CSEI performed the anti-hepatocarcinoma activity via ROS / MAPK-dependent mitochondrial pathway.3.Study on the immunomodulatory activity of CSEIIn this paper,the immunomodulatory activity of CSEI was investigated by establishing an immunosuppressive model induced by cyclophosphamide(CTX).After 14 days of treatment,CSEI reversed the CTX-induced immunosuppression,and significantly increased spleen index and restored serum IL-2,6,10,12 and IFN-?,? and TNF-? and other immune and inflammatory cytokine levels,and increased NK cell activity and T lymphocyte transformation ability.In addition,the phosphorylation of IKK? in spleen was increased.In summary,CSEI is a safe and effective anti-hepatoma drug,and also performs immunomodulatory effect and proapoptosic effect in human hepatoma cell via ROS / MAPKs-dependent mitochondrial pathway.The present study provides pharmacological evidence for CSEI as a potential therapeutic agent for hepatocellular carcinoma and other primary tumors.