KIAA1429 Regulates EZH2 And DNMT1 RNA Methylation To Promotes Alcohol-associated Hepatocellular Carcinoma Progression

Background:Liver cancer is one of the most common malignant tumors and the fourth most lethal tumor,with over 850,000 people diagnosed each year.Hepatocellular carcinoma(HCC)accounts for about 90%of liver cancer cases.Viral infection,alcoholism,and metabolic syndrome are the most common risk factors for liver cancer.Alcohol consumption is linked to a high mortality rate and a poor prognosis of HCC.N6-methyladenosine(m6A)methylation is the most common post-transcriptional modification of oncogenes in tumors.In HCC,abnormal activation of M6A-related proteins regulates cell proliferation,invasion,and epithelial mesenchymal transformation,promoting the progression of the disease.However,it is unknown how m6A methylation contributes to the development of alcohol-related hepatocellular carcinoma(A-HCC).Simultaneously,the relevant targets and mechanisms of m6A regulation of A-HCC progression remain unclear.Therefore,we attempted to provide new evidence for improving the prognosis of A-HCC patients by focusing on the role of m6A methylation in A-HCC progression and related mechanisms.Methods:1.To construct risk assessment models,public databases were used to collect RNA sequencing data and clinical information from A-HCC patients,and m6A regulatory factors closely related to the prognosis of A-HCC were screened out using STRING and Cbioportal tools;2.The expression of m6A-related factor RNA and protein in clinical tissue samples was confirmed using fluorescence quantitative PCR and immunohistochemistry,and the key m6A genes influencing the prognosis of AHCC were identified using survival analysis;3.To identify the target of m6A methylation affecting A-HCC progression,lentivirus was transfected into Huh7,HepG2 and Hep 1-6 cells,followed by subcutaneous tumorigenosis and in situ liver implantation experiments;4.The CTRP database was used to screen potential therapeutic drugs acting on corresponding targets,and in vivo and in vitro A-HCC models were created.Immunostaining and Western blot analysis were used to investigate changes in m6A methylation levels,and the therapeutic effects of potential drugs on A-HCC were evaluated.Results:1.The expression of m6A-related regulatory factors LRPPRC,YTHDF2,KIAA14219,and RBM15B was significantly increased in A-HCC patients,and the m6A risk assessment model was developed to divide A-HCC patients into high and low risk groups;2.The prognosis of A-HCC disease was worse in the m6A high-risk group than in the low-risk group;3.Pathway enrichment revealed that pathways related to tumorigenesis and proliferation,such as DNA repair and the mTOR pathway,were activated in the high-risk A-HCC group;4.The high expression of the m6A gene KIAA1429 is associated with a poor prognosis for patients.The ability of the cell to proliferate,invade,and migrate is significantly reduced after KIAA 1429 knockdown in Huh7 and HepG2,and the survival time of mice with orthotopic HCC cell implantation was significantly extende;5.KIAA1429 knockdown can significantly shorten the half-life of EZH2 and DNMT1 transcript degradation,decrease mRNA stability,and reduce protein expression levels;6.Tenipside inhibits KIAA1429,which lowers the expression of EZH2 and DNMT1,and inhibits the expression of Wnt/βcatenin signaling related genes.This reduces the number and size of tumors in A-HCC mouse models and improving prognosis.Conclusion:The m6A risk model accurately predicts the prognosis of A-HCC.The m6A regulatory factor KIAA1429,a key factor affecting A-HCC patients,promotes the progression of A-HCC by regulating the expression of EZH2 and DNMT1.Teniposide,a potential A-HCC therapeutic drug,can reduce the mRNA stability of EZH2 and DNMT1 by down-regulating the expression of KIAA1429.This reduces the protein expression of EZH2 and DNMT1,inhibiting the activation of Wnt/β-catenin signaling pathway and improves the prognosis of A-HCC.