Anti-liver Cancer Activity And Mechanism Of Glycycoumarin

Licorice is a traditional Chinese medicine.It has been used in treating various diseases for centuries,including liver disease.However,the chemical basis or biological mechanisms underlying the biological functions of licorice has not been scientifically addresssed.Glycycoumarin(GCM)is a major bioactive coumarin compound isolated from licorice.The anti-cancer activity of GCM remain elusive.In this study,we have demonstrated the anti-liver cancer activity and the mechanism of GCM using different models.Primary liver cancer(PLC),including hepatocellular carcinoma(HCC)and intrahepatic cholangiocarcinoma(ICC),is one of the most common malignancies and the second leading cause of cancer associated mortality worldwide.PLC is a well known chemoresistant tumor,making them among the most lethal cancers without effective treatment options.Therefore,there is an urgent need to develop novel therapeutic strategies for the treatment of this lethal malignancy.In the present study,using animal models of both chronic and acute alcoholic liver injury,we demonstrated the role of Glycycoumarin in alcohol-induced hepatotoxicity.C57BL/6J mice were used to evaluate the hepatoprotective effect of Glycycoumarin on animal models of liver injury.AML-12 and HepG2 cells were utilized to determine the functional role of Nrf2 in the hepatoprotective effect of Glycycoumarin and to decipher the mechanisms of Glycycoumarin-induced Nrf2 activation.We found that treatment with Glycycoumarin leads to a significant reduction in hepatotoxicity in response to either chronic or acute ethanol exposure.Further mechanistic investigations reveal that activation of Nrf2 via the p3 8 pathway and induction of autophagy by Glycycoumarin contribute to its hepatoprotective activity.This effect of Glycycoumarin reduces the risk of the conversion of alcoholic fatty liver to liver cancer.Next,we tested the anti-liver cancer activity of Glycycoumarin using both in vitro and in vivo models and found for the first time that Glycycoumarin possesses a potent activity against liver cancer evidenced by cell growth inhibition and apoptosis induction in vitro and tumor reduction in vivo.Mechanistically,Glycycoumarin was able to bind to and inactivate oncogenic kinase T-LAK cell-originated protein kinase(TOPK),which in turn led to activation of p53 pathway.Our findings supported Glycycoumarin as a novel active compound that contributed to the anti-cancer activity of licorice and TOPK could be an effective target for hepatocellular carcinoma(HCC)treatment.We also foundGlycycoumarin can significantly inhibit the mTOR activity of hepatocellular carcinoma or intrahepatic cholangiocarcinoma and cause cell cycle arrest.Based on this study,we also illustrated the therapeutic effects of mTOR inhibitor and CDK4/6 inhibitor alone or combined treatment in AKT/Yap induced ICC in mice.We show for the first time that combining pan-mTOR inhibitor and CDK4/6 inhibitor synergies to restrain the growth of ICC cells in vitro.Importantly,while pan-mTOR inhibitor or CDK4/6 inhibitor alone shows moderate efficacy in a genetic mouse ICC model,the combined treatment leads to tumor regression in mice.Mechanistically,we demonstrate that combined treatment with pan-mTOR and CDK4/6 inhibitors results in strong inhibition of cell proliferation in ICC cells in vitro and in vivo.CDK4/6 inhibitor potentiates mTOR inhibitor's activity;whereas mTOR inhibitor prevents CDK4/6 inhibitor induced upregulation of cyclin D1.This article focuses on the prevention and treatment of primary liver cancer,clarifing the efficacy of Glycycoumarin on alcoholic liver injury,hepatocellular carcinoma and intrahepatic cholangiocarcinoma.Meanwhile,the cell cycle inhibition and mTOR signaling pathway decreasion effect of Glycycoumarin were further studied to elucidate the anticancer effect of inhibiting the corresponding target.In summary,this paper provides a new treatment method and theoretical basis for the prevention and treatment of liver cancer.