NENF, as a Biomarker for the Development of NAFLD to Liver Cancer by Enhancing the Tumor‐Like Stem Cell Properties of L02 Cells to Promote the Progression of NAFLD to Liver Cancer

ABSTRACT To explore the key risk genes involved in developing NAFLD into HCC. Four datasets‐related NAFLD progression (NAFLD, NASH, Hepatofibrosis, Cirrhotic, and Tumor) were obtained from the GEO database. GO and KEGG analyses were performed to identify the biological functions, pathways, and cellular processes associated with the genes. GSVA analyses were performed to assess the variation in pathway activity across different samples based on gene expression profiles. We performed ordinal logistic regression analysis, collinearity analysis, importance analysis of influence factors (LASSO, XGBoost, and RandomForest), and Venn analysis to identify the hub genes within disease progression. The SHAP model and K‐M survival analysis screened out risk genes‐related development of NAFLD into HCC. The mRNAsi_score analysis evaluates the correlation of NENF expression and tumor stemness. A vector expressing NENF transfection was used to increase its expression. CCK8, cell spheroid formation assay, colony formation, transwell, western blot, and RT‐PCR assays were used to detect cell viability, tumor stemness, and gene and protein expression. NENF high expression was associated with NAFLD progression NENF was identified as the risk gene and positively associated with mRNAsi_score. Its expressions gradually increased with the progress of NAFLD to HCC. NENF promoted cells' lipid accumulation. NENF enhanced the induction of L02 cells into stem cells and augmented the tumor‐like stem cell properties of L02 cells. NENF, as an important biomarker, promoted the development of NAFLD devolved to HCC by enhancing the tumor‐like stem cell properties of normal cells.