KAT2A Promotes Proliferation,migration And Invasion Of Hepatocellular Carcinoma Cells Through PI3K/AKT Signaling Pathway

Aims: Lysine Acetyltransferase 2A(KAT2A),as one of the important regulatory enzymes of protein acetylation modification,is involved in the occurrence and development of various malignant tumors.The objective of this study was to investigate the effects of KAT2 A on the proliferation,migration and invasion of Hepatocellular carcinoma(HCC)cells and its mediated signaling pathway.Methods:(1)To explore the relationship between KAT2 A and HCC from multiple perspectives based on bioinformatics.Firstly,we analyzed the expression of KAT2 A m RNA and protein in HCC tumor tissues and normal tissues using The Cancer Genome Atlas(TCGA)database and UALCAN database,and analyzed the correlation between KAT2 A and different clinical indices of HCC patients based on the clinical information in the database.Then the GEPIA database and Kaplan-Meier Plotter database were used to explore the effects of different expression of KAT2 A m RNA on different survival of HCC patients,and then further explore the effects of different clinical indicators on the overall survival of HCC patients and perform risk factor analysis.Finally,ROC curves were drawn using KAT2 A as a diagnostic predictor and prognostic predictor.The c Bio Portal database was used to investigate the cause of high KAT2 A expression in HCC.TIMER,TIMER2.0 databases were used to analyze the relationship between KAT2 A and immune cell infiltration in HCC.The TCGA data and STRING database were used for the functional investigation of KAT2 A,and the cellular components,molecular functions,biological processes,and pathways involved in KAT2A-associated genes,as well as the interacting proteins of KAT2 A were obtained,respectively.(2)The role and mechanism of KAT2 A in HCC were further investigated by in vitro cellular experiments.The expression levels of KAT2 A m RNA and KAT2 A protein in different HCC cells and the constructed HCC cells with low KAT2 A expression were examined using Real time quantitative polymerase chain reaction(RT-q PCR)and protein immunoblotting(Western blot,WB),respectively.The CCK8 assay,clone formation assay,scratch assay,invasion assay and phosphatidylinositol 3 kinase serine/protein kinase B(PI3K-AKT)pathway assay were then used to investigate the effects of KAT2 A on the proliferation,migration and invasion of HCC cells and the potential pathways of action.Results:(1)Bioinformatic analysis showed that KAT2 A m RNA expression was elevated in 15 cancers including HCC.protein expression of KAT2 A was higher in HCC tumor tissues than in normal tissues.logistic regression analysis suggested that the main factors affecting KAT2 A expression in HCC were body weight,prolonged prothrombin time,alpha fetoprotein(AFP),vascular invasion,T stage,pathological stage,and histological grade.KAT2 A affects the survival of HCC patients,including overall survival(OS),progression free survival(PFS)and relapse free survival(RFS).KAT2 A had a significant effect on OS in HCC patients with different clinical indicators,including tumor stage T3 group,pathological stage III group,histological grade G1 group,no vascular invasion group,treatment with sorafenib group,no hepatitis group and alcohol consumption group.The methylation level of KAT2 A was negatively correlated with its m RNA expression level.k AT2 A was able to influence the infiltration level of immune cells in HCC patients and was positively correlated with tumor-promoting myeloid-derived suppressor cells(MDSC)and regulatory T(Treg)cells.GO and KEGG analyses showed that KAT2A-related genes were mainly enriched in transcriptional regulation,cell cycle and cell division.Interacting protein analysis showed that KAT2 A interacting proteins were involved in transcriptional complex formation and pro-tumorigenic development,including MYC,TADA3,CCDC101,TRRAP,TADA2 A,TADA2B,USP22,ATXN7,and TAF10.(2)In vitro cellular experiments showed that KAT2 A m RNA and protein expression were higher in HCC Huh7 and SK-hep-1 cell lines.low expression of KAT2 A in HCC cells inhibited tumor cell proliferation,clone formation,migration and invasion,and decreased activation of the PI3K/AKT pathway.Conclusions:(1)Bioinformatics showed that KAT2 A m RNA and protein were highly expressed in HCC tissues,and high KAT2 A m RNA expression was associated with shorter survival in HCC patients,and KAT2 A may be a potential prognostic marker in HCC patients.(2)I In vitro cell experiments demonstrated that the low expression of KAT2 A blocked the activation of PI3K/AKT signaling pathway,and significantly weakened the proliferation,migration,and invasion of SK-hep-1 and Huh-7 cells of HCC.