Hypoxia induced DNMT3B and SHP2 signaling promoted HCC via suppressing P53 and MYH11 protein expression
Source: PubMed Central Open Access, NCBI / U.S. National Library of Medicine
Objective This work aims to analyze the intricate process via which CBFβ-MYH11 facilitates the suppression of hepatocellular carcinoma in hypoxic environments. Methods Three independent HCC cohorts including TCGA-LIHC,and ICGC LIRI-JP were enrolled. Gene correlation analysis, hypoxia-based expression comparison and prognostic stratification analysis were performed. A composite HIF1A/AKT/SHP2/DNMT3B/P53/MYH11 axis score was established to evaluate clinical prognostic value. Bioinformatics analysis was utilized to examine differentially expressed genes in hepatocellular carcinoma. In nude mouse models, tumor volume, tumor cross-sectional area, methylation levels, protein expression, vascular development, scratch assay, invasion assay, chromatin immunoprecipitation, and co-immunoprecipitation were quantitatively assessed. We used methods including HE staining, immunohistochemistry, phosphor-PCR, Western blot, angiogenesis tests, migration assays, and invasion assays. Results Multi-cohort analysis revealed a stable negative correlation between DNMT3B and MYH11. High HIF1A expression was accompanied by DNMT3B upregulation and MYH11 silencing. Patients in the high axis score group exhibited markedly shorter overall survival, and the axis score served as an independent adverse prognostic risk factor. Bioinformatics analysis reveals MYH11-specific expression in hepatocellular carcinoma. There were clear differences between Huh-7-NC-CBFβ-MYH11 and Huh-7-OE-CBFβ-MYH11 when they were co
Abstract
Objective This work aims to analyze the intricate process via which CBFβ-MYH11 facilitates the suppression of hepatocellular carcinoma in hypoxic environments. Methods Three independent HCC cohorts including TCGA-LIHC,and ICGC LIRI-JP were enrolled. Gene correlation analysis, hypoxia-based expression comparison and prognostic stratification analysis were performed. A composite HIF1A/AKT/SHP2/DNMT3B/P53/MYH11 axis score was established to evaluate clinical prognostic value. Bioinformatics analysis was utilized to examine differentially expressed genes in hepatocellular carcinoma. In nude mouse models, tumor volume, tumor cross-sectional area, methylation levels, protein expression, vascular development, scratch assay, invasion assay, chromatin immunoprecipitation, and co-immunoprecipitation were quantitatively assessed. We used methods including HE staining, immunohistochemistry, phosphor-PCR, Western blot, angiogenesis tests, migration assays, and invasion assays. Results Multi-cohort analysis revealed a stable negative correlation between DNMT3B and MYH11. High HIF1A expression was accompanied by DNMT3B upregulation and MYH11 silencing. Patients in the high axis score group exhibited markedly shorter overall survival, and the axis score served as an independent adverse prognostic risk factor. Bioinformatics analysis reveals MYH11-specific expression in hepatocellular carcinoma. There were clear differences between Huh-7-NC-CBFβ-MYH11 and Huh-7-OE-CBFβ-MYH11 when they were compared. Huh-7-OE-CBFβ-MYH11 showed a smaller tumor volume, less vascular density, fewer invasion cells, a shorter migration distance, and less vascular development. The expression of MMP2, VEGF, and HIF1α showed a decrease, while RUNX1 expression increased. The introduction of an AKT inhibitor in hypoxic conditions resulted in a more pronounced increase in P53 and CBFβ-MYH11 protein expression compared to the SHP2 inhibitor. AKT inhibitors eliminated differences in DNMT3B, HIF1α, and MMP2 expression between normoxia and hypoxia. Conclusion In hypoxic conditions, CBFβ-MYH11 regulates the AKT/DNMT3B/SHP2 pathway to modulate variations in P53 expression, ultimately hindering the progression of hepatocellular carcinoma.