Effect Of Bufalin Combined With Sorafenib Of Hepatoma Cell

?Background?Hepatocellular carcinoma(HCC)accounts for 80 to 90 per cent of primary liver cancer.Hepatocellular carcinoma(HCC)is one of the refractory malignant tumors with the highest mortality rate.The mortality of HCC in China ranks the second place after lung cancer.Liver cancer is one of the most common malignant tumors in the world.Compared with developed countries in Europe and America.,most liver cancer patients in China,their treatment means are very limited,accompanied by blood metastasis and tumor thrombogenesis,which directly leads to a low five-year survival rate of liver cancer.Therefore,how to treat liver cancer and improve the prognosis of liver cancer surgery is particularly important.Traditional chemotherapy for liver cancer is generally used clinically as chemotherapy drugs,such clinical treatment often develops into chemical resistance.Sorafenib,for example,is a multi-kinase inhibitor.Its mechanism of action is mainly through inhibiting the generation of tumor blood vessels,which has achieved a good effect in the treatment of advanced liver cancer and can significantly prolong the survival time of patients with advanced liver cancer.Sorafenib is used as chemotherapy for advanced liver cancer.After treatment with sorafenib,some patients develop resistance to it.Sorafenib as SOR drug is one of the difficult problems in anticancer therapy.To study the mechanism of sorafenib resistance and improve its efficacy has become the focus of sorafenib research.Recent studies have shown that many active ingredients extracted from Chinese herbal medicine have good anti-tumor potential.Bufalin is the dried secretions of the glands behind the ears from Toad cake of the Toads family.Bufalin can inhibit the activity of topoisomase 11 and interfere with cell DNA replication.Most early studies focused on inducing the differentiation and apoptosis of human leukemia.In recent years,studies have shown that bufalin can inhibit the growth of endometriosis cells,ovarian cancer and prostate cancer.It has been reported that bufalin and sorafenib can combine with m TOR and VEGF in a good anti-cancer effect.In this paper,the inhibitory effect of bufalin combined with sorafenib on the proliferation of liver cancer was studied and the possible molecular mechanism was discussed.?Objective?To explore the inhibitory effect of bufalin combined with sorafenib on the proliferation of hepatocellular carcinoma cells SNU-739 and its related molecular mechanism through in vitro and in vivo experiments?Methods?1.The inhibitory effects of different concentrations of bufalin,sorafenib and sorafenib combined with sorafenib on the proliferation of hepatocellular carcinoma cells were detected by MTT,cloning and formation experiments,LDH generation detection,scratching and microscopy2.The damage of sorafenib on normal liver cells can be reduced by using MTT method,clone formation experiment,LDH generation detection,scratch and microscopic examination of bufalin.(the experiment was divided into control group,bufalin group,sorafenib group,and bufalin + sorafenib group)3.Flow,DAPI,q PCR,WB,TUNEl,EDU and other methods were used to study the effect of bufalin on the apoptosis of liver cancer cells.4.Mitochondrial membrane potential jc-1,ATP generation study,mitochondrial copy number bufalin enhanced sorafenib through mitochondrial pathway to cause liver cancer cell death.5.Tumor xenotransplantation and immunohistochemical experiments were conducted to verify the anti-tumor effect of bufalin on sorafenib in mice.?Results?(Part I)1.Bufalin inhibits proliferation and migration of liver cancer cells,and IC50 is calculated as 500 n M.0,500,1000 and 2000 n M were selected as experimental drug concentrations.(Part II)1.The inhibitory effect of bufalin combined with sorafenib on liver cancer cells was significantly greater than that of bufalin and sorafenib alone.2.Bufalin and Sorafenib inhibited the proliferation of liver cancer in nude mice.(Part III)1.Bufalin combined with sorafenib can significantly induce the apoptosis of HEPATocellular carcinoma cells SNU-739.2.Bufalin combined with sorafenib induced apoptosis of HCC cells SNU-739 through mitochondria-related pathways,and inhibited proliferation of hepatocellular carcinoma cells SNU-739.?Conclusion?The above experimental results indicate that bufalin can enhance the inhibitory effect of sorafenib on liver cancer cells and enhance the apoptosis of liver cancer cells by affecting the mitochondria of liver cancer cells.And bufalin can reduce the effect of sorafenib on normal liver cells.At the same time,in vivo experiments in nude mice confirmed that bufalin can significantly enhance the anti-tumor effect of sorafenib in vitro.