Study On The Effect Of Anti-hepatoma Of Nigericin And Aerobic Glycolysis

Hepatocellular carcinoma poses a serious threat to human health.Patients with advanced liver cancer usually need to receive anti-tumor drug treatment.The therapeutic effects of existing chemotherapy drugs and molecular targeted drugs on hepatocellular carcinoma in clinical practice are not ideal,and there is an urgent need to develop effective and stable new drugs.Nigericin sodium salt(NSS)is a polyether antibiotic derived from Streptomyces hygroscopicus.It has been found that NSS has effective anti-tumor activity against colon cancer,breast cancer,lung cancer in vivo and in vitro studies.There is limited research on the molecular mechanisms underlying the anti-tumor effect of NSS,and further research is needed.Hepatocellular carcinoma cells,like the tumor cells from other tissues,also undergo changes in energy metabolism.The glucose metabolism of tumor cells is usually converted to aerobic glycolysis from the process of oxidizing glucose.The tumor cells mainly rely on aerobic glycolysis for energy supply.Many key enzymes and regulatory proteins in the glycolytic pathway play a crucial role in tumor development.Aerobic glycolysis is related to the proliferation,immune escape,invasion,migration,and angiogenesis of hepatocellular carcinoma.On the basis of exploring the anti-hepatoma effects of NSS in vivo and in vitro,this study takes aerobic glycolysis as the breakthrough point to clarify the effect of NSS on aerobic glycolysis of hepatocellular carcinoma and its molecular mechanism.Experiments in vitro were respectively conducted to detect the proliferation,apoptosis,migration,and invasion ability of liver cancer cells by MTT assay,Annexin V-FITC and PI double staining,scratch test,and Transwell invasion assay.The content of lactic acid(LD)in the supernatant of hepatocellular carcinoma cell culture and the activities of hexokinase(HK),lactate dehydrogenase(LDH),and pyruvate kinase(PK)in the cells were measured by biochemical methods.Western blot was used to detect the expression level of apoptosis related proteins Bax,Bcl-2,glycolysis related regulatory proteins of AMPK-P,AMPK,GSK-3 and Glut1 in hepatocellular carcinoma cells.In the experiments in vivo,Hepa1-6 bearing mice subcutaneous transplanted tumor model were established,and were divided into model control group(physiological saline),DOX group(0.05mg/kg),and NSS group(0.05mg/kg,0.1mg/kg).NSS and DOX were injected into transplanted tumor.The tumor inhibition rates were calculated.The lactate content,activities of key glycolytic enzyme-HK,LDH,PK in the tumor tissue were measured.The main research results of this study include:1.The effect of NSS on the biological characteristics of hepatocellular carcinoma cells NSS can significantly inhibit the proliferation of hepatocellular carcinoma cells,induce apoptosis in some hepatocellular carcinoma cells,upregulate Bax protein expression,and downregulate Bcl-2 protein expression,significantly reducing cell migration and invasion ability.2.Study on the effect of NSS on aerobic glycolysis of hepatocellular carcinoma cells NSS can significantly reduce LD content in the culture supernatant of hepatocellular carcinoma cells and the activity of HK,LDH,PK in these cells;and it also significantly downregulate the protein expression level of activated AMPK,GSK-3β.3.The anti-tumor activity and the effect on aerobic glycolysis of NSS on Hepa1-6tumor bearing mice Compared with the model group,the tumor weight significantly decreased and the tumor inhibition rates significantly increased in NSS groups;NSS significantly reduced the lactate content and inhibited the activity of key glycolytic enzymes-HK,LDH,and PK in tumor tissue.In summary,NSS has strong anti-hepatoma activity,can induce cell apoptosis,and inhibit the migration and invasion ability of hepatocellular carcinoma cells;Inhibiting the aerobic glycolysis and affecting its key enzyme activity as well as the expression of regulating protein in hepatocellular carcinoma cells may be an important molecular mechanism of anti-hepatoma effect of NSS.This study lays the experimental foundation for further exploring the anti-tumor effects of NSS on metabolic pathways.