AFAP1-AS1/miR-195-5p is associated with malignant phenotypes and glycolysis-related metabolic changes in hepatocellular carcinoma

Abstract The long noncoding RNA AFAP1-AS1 is aberrantly expressed in multiple solid tumors, but its functional role in hepatocellular carcinoma (HCC) remains incompletely characterized. In this study, analysis of the TCGA-LIHC cohort and eight paired HCC and matched adjacent non-tumor tissues showed that AFAP1-AS1 was upregulated in HCC and associated with poorer overall survival in the TCGA cohort. Knockdown of AFAP1-AS1 inhibited the proliferation, migration, and invasion of HCC cells. Bioinformatic prediction, expression analysis, and dual-luciferase reporter assays supported a possible regulatory relationship between AFAP1-AS1 and miR-195-5p at the predicted binding site. Inhibition of miR-195-5p increased FOXK1 expression, consistent with the effectiveness of miR-195-5p inhibition, and enhanced HCC cell migration and invasion. It also increased glucose consumption and lactate production, indicating glycolysis-related metabolic changes. Exploratory RNA-seq analysis after miR-195-5p inhibition, integrated with FOXK1 target genes and a glycolysis-related gene set, identified four candidate genes. However, these candidates and the role of FOXK1 in mediating the observed phenotypes were not functionally validated. Rescue experiments showed that miR-195-5p inhibition partially reversed the suppressive effects of AFAP1-AS1 knockdown on HCC cell proliferation, migration, and invasion. In addition, inhibition of miR-195-5p promoted tumor growth in a HCC xenograft model. Overall, these findings suggest a possible association of AFAP1-AS1 and miR-195-5p with malignant phenotypes and glycolysis-related metabolic changes in HCC.