The Function Of E3 Ubiquitin Ligase HERC1 In Hepatocellular Carcinoma Tumorigenesis And Metastasis

Background :Liver cancer is one of the 10 most common malignancies in the world,having the insidiou he rapid disease progression.When the typical symptoms and signs appear,it often comes into the advanced stage.Surgical treatment for liver cancer is the most important ways for liver cancer patients to get the long-term survival,but its recurrence rate is as high as 40% ~ 70% after surgical resection.In recent years,there has been great progress in the clinical application of molecular targeted drugs.Molecular targeted drug sorafenib has certain benefits for advanced liver cancer patients with different liver disease backgrounds.This provides a way for our scientific research to find new drug targets for diagnosis and treatment of liver cancer.Previously,we found that the ECD inhibited the ubiquitination of hnRNPK to up-regulate its protein expression level,thereby promoting Hepatocellular carcinoma(HCC)cell proliferation,cloning formation,invasion and metastasis.In order to find out the hiding mechanism,we found out ubiquitin ligase E3,HERC1 which regulates ubiquitination degradation of hnRNPK,by mass spectrometry asssay.Therefore,this acticle mainly focuses on the role of HERC1 in HCC and its clinical significance.Investigating the molecular mechanism of HERC1 involved in regulation,we hope to provide the basis for targeted therapy of HCC.Methods and Results:1.The interaction between ECD protein and hnRNP K protein was confirmed byCo-IP and Western blotting.2.The interaction between hnRNP K protein and HERC1 protein was confirmed byCo-IP and Western blotting.3.The Co-IP and Western blot confirmed that ECD protein hinders the interactionbetween hnRNP K protein and HERC1 protein.4.Agarose gel electrophoresis confirmed that herc1 did not affect the expression ofhnrnpk at rna level.5.The Western blot confirmed that si-herc1 upregulated the expression of hnrnpk atprotein translation level.6.The Western blot confirmed that si-herc1 enhanced the stability of hnrnp kprotein at protein translation level.7.The Western blot confirmed that si-herc1 inhibited the ubiquitin degradation ofhnrnp k protein.8.The Western blot confirmed that ov-ECD and si-herc1 inhibited the degradationof hnrnp k protein.9.Through using of Western blotting,the proliferation experiment,clone formingassay,Transwell chamber and other methods,we confirmed that si-ECD inhibitscell proliferation,colony formation,migration and invasion.Conclusion:1.In vitro experiments confirmed that si-ECD can promote the proliferation,cloneformation,invasion and metastasis of hepatocellular carcinoma cells.2.ECD promotes the proliferation,clone formation,invasion and metastasis ofhepatocellular carcinoma cells by blocking E3 ligase HERC1-mediatedhnRNPK ubiquitination and degradation.