Study On The Role And Mechanism Of UBE2L3 In Hepatocellular Carcinoma

Background & ObjectiveHepatocellular carcinoma(HCC)is one of the most common malignancies in the world,its morbidity and mortality are increasing in both developed and developing countries.The occurrence and development of hepatocellular carcinoma(HCC)is a multi factor,multi-level,multi-step process,which is affected by environmental factors,viral infection,host genetic variation and other factors.Among them,chronic hepatitis B virus(HBV)infection is the most important risk factor of HCC,in our country about 85% to 90% of HCC are associated with hepatitis B virus infection.At present,due to the lack of early diagnosis and effective treatment,HCC patients with poor prognosis,five-year survival rate is low.Therefore,it is of great significance to explore the molecular pathological mechanism of the occurrence and development of HCC and search for effective therapeutic targets.UBE2L3(Ubiquitin-conjugating enzyme E2L3),also known as UbcH7,belongs to the ubiquitin-binding enzyme E2,which participates in the ubiquitination process of protein and mediates the ubiquitination of c-Fos,p53,NF-kB precursor p105 in vitro,associated with rheumatoid arthritis,Crohn's disease,systemic lupus erythematosus and psoriatic arthritis and other autoimmune diseases.Recent genome-wide association studies show that UBE2L3 is a susceptibility gene of chronic HBV infection,chronic HBV infection as the most important risk factor of HCC,the effect and significance of susceptible gene locus on HBV-related hepatocellular carcinoma need to be studied.Therefore this experiment through large multicenter case-control study,we analyzed the association between UBE2L3 genetic variation and the susceptibility to HBV-related hepatocellular carcinoma(HCC),the role and mechanism of UBE2L3 in the occurrence and development of hepatocellular carcinoma were further studied by animal model test,clinical liver cancer tissue analysis and detection of biological characteristics of hepatocellular carcinoma cells in vitro.Methods1.The correlation between UBE2L3 genetic variation and susceptibility to HBV-related hepatocellular carcinoma was analyzed by a large sample of multicenter case-control studies.2.The relationship between the expression level of UBE2L3 and the malignancy and survival rate of hepatocellular carcinoma was analyzed according to the information of The Cancer Gene Atlas(TCGA)and clinical hepatocellular carcinoma tissue array.3.The expression of UBE2L3 in human hepatocellular carcinoma specimens was detected by immunohistochemistry.The expression of UBE2L3 in human hepatoma cells was detected by Western-Blot and RT-PCR.4.The proliferation ability of the cells was detected by CCK-8 and plate colony forming assay,and the migration ability of the cells was detected by Transwell chamber assay.5.Using lentiviral vector to construct stable hepatoma cell lines that knockdown UBE2L3,RNA sequencing was used to study the effect of UBE2L3 knockdown on the occurrence and development of hepatocellular carcinoma.Result1.Genetic variation of UBE2L3 is associated with HCC and chronic HBV infection2.The expression of UBE2L3 in clinical hepatocellular carcinoma tissues was significantly higher than that in adjacent normal tissues,and the expression of UBE2L3 is also positively correlated with tumor grade,the higher grade of hepatocellular carcinoma,the higher expression of UBE2L3 was observed.Besides,hepatocellular carcinoma patients with higher expression of UBE2L3 demonstrate poor survival.3.In vitro experiments showed that knockdown of UBE2L3 could decrease the proliferation and migration ability of hepatoma cells,while overexpression of UBE2L3 could increase the proliferation and migration of hepatoma cells.4.UBE2L3 can regulate the EMT process,and negatively regulate the expression of CLDN1 and CDKN2 B two tumor suppressor genes to promote the occurrence and development of hepatocellular carcinoma.