The Mechanism Of BARX2 Inhibiting The Proliferation,Invasion And Migration Of Hepatocellular Carcinoma By Activating GALNT4

Objective:BARX2(Bar H-like homeobox 2),a member of the BAR family of homology domain transcription factors,is biologically important in cell adhesion,embryonic development,and especially in the regulation of tumor cell proliferation and metastasis.Some studies have found that BARX2 has been identified to regulate the development of multiple cancers and is associated with patient prognosis,suggesting that BARX2 may be an independent prognostic biomarker for patients with hepatocellular carcinoma.However,the biological function of BARX2 in hepatocellular carcinoma remains in question and requires further scientific exploration.Aims of this study were to investigate the expression level of BARX2 in hepatic celluler carcinoma and the clinical significance of its abnormal expression,and to explore the mechanism by which BARX2 regulates the peptide N-acetylgalactosaminyltransferase 4(GALNT4)to affect the proliferation and invasion and migration of hepatocellular carcinoma.Methods:1.Analysis of BARX2 expression in hepatocellular carcinoma and its association with patient survival prognosis by bioinformatics approach using TCGA database;2.The expression of BARX2 and N-acetylgalactosaminyl transferase 4(GALNT4)in hepatocellular carcinoma tissues and normal tissues adjacent to the carcinoma were detected by real-time fluorescence quantitative polymerase chain reaction(q RT-PCR)and protein blotting(Western blot)at the m RNA level and protein level,respectively;3.Interfering and overexpressing BARX2 cell lines were evaluated by CCK-8proliferation assay,colony formation assay,scratch test,transwell invasion experiment and angiogenesis assay,respectively,in terms of cell proliferation,invasion,migration and angiogenesis ability;4.The binding relationship between BARX2 and GALNT4 was analyzed through the JASPAR website and validated using chromatin;5.immunoprecipitation(Ch IP)and luciferase reporter assays.Results:1.Analysis of BARX2 expression in hepatocellular carcinoma,using hepatocellular carcinoma data from the TCGA database,revealed that BARX2 expression was higher in normal tissue adjacent to cancer than in hepatocellular carcinoma tissue and that its low expression was an independent risk factor for poor prognosis in patients with hepatocellular carcinoma;2.BARX2 expression was significantly reduced in hepatocellular carcinoma tissues as well as in hepatocellular carcinoma cell lines;3.Cell viability and cell colony formation were significantly decreased after BARX2 overexpression in hepatocellular carcinoma cell lines;the expression level of GALNT4 in BARX2 overexpressing hepatocellular carcinoma cell lines was detected,and BARX2 overexpression significantly increased the level of GALNT4;4.BARX2 exerts anti-hepatocellular carcinoma activity by activating GALNT4,and is able to bind to the GALNT4 promoter and positively regulate GALNT4expression;5.GALNT4 plays a key effector molecular role in the regulation of multiplication,invasiveness and immigration of liver cancer cells by BARX2.Conclusion:1.Aberrant low expression or loss of BARX2 expression in hepatocellular carcinoma tissues may promote proliferation,invasion and migration,and low level of BARX2 expression may suggest metastasis of hepatocellular carcinoma;2.BARX2 has the potential to be a new molecular marker for predicting the prognosis of patients with hepatocellular carcinoma and may be a new target for the treatment of hepatocellular carcinoma;3.BARX2 positively regulates GALNT4 expression by binding to the GALNT4 promoter and inhibits the proliferative capacity of hepatocellular carcinoma cells.