Par-4/TERT induced autophagy dysregulation in regulating glycolysis in hyperglycemia combined with hepatocellular carcinoma.

To explore the mechanism by which the Prostate apoptosis response-4 (Par-4)/Telomerase Reverse Transcriptase (TERT) axis induces tumor progression by regulating autophagy and glycolysis in hepatocellular carcinoma (HCC) under hyperglycemic conditions. The high-glucose Hepa1-6 cell model and nude mouse HCC xenograft model were used, along with gene knockdown (shPar-4, shTERT) and overexpression (oePar-4) strategies. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, flow cytometry, Western blot, TUNEL staining, co-immunoprecipitation (co-IP), quantitative real-time PCR (qPCR), and animal experiments were performed to analyze the effects of Par-4/TERT on AKT activity, autophagic activity, and the expression of key glycolytic enzymes (GLUT-1, HK, PFK, and PK). The anti-tumor effects of AKT inhibitors (SH-5 and LY294002), the autophagy regulator (rapamycin), and the glycolysis inhibitor (2-deoxy-D-glucose, 2-DG) were further evaluated. Under hyperglycemic conditions, Par-4 expression was significantly suppressed, whereas TERT expression was markedly upregulated. This elevated TERT expression activated the AKT/NF-κB/FOXO3a signaling pathway, leading to enhanced glycolytic metabolism (evidenced by upregulated expression of glucose transporters such as GLUT-1 and key glycolytic enzymes) and induced autophagic processes (characterized by increased LC3-II/LC3-I ratios and decreased p62 levels). Par-4 overexpression or TERT knockdown effectively reversed these effects, resulting in inhibited cell proliferation and induced apoptosis. Co-immunoprecipitation (co-IP) experiments confirmed a direct interaction between Par-4 and TERT, with Par-4 negatively regulating TERT expression. In combination therapeutic approaches, the LY294002 + 2-DG regimen exhibited synergistic anti-tumor efficacy in models with Par-4 overexpression or TERT knockdown, achieving a greater than 50% reduction in tumor volume accompanied by downregulated p62 expression. The Par-4/TERT regulatory axis modulates autophagy dysregulation and glycolytic metabolism through the AKT signaling cascade, contributing to HCC progression under hyperglycemic conditions. Synergistic targeting of this axis, in conjunction with AKT blockade and glycolytic inhibition, exerts potent anti-tumor activity, providing a novel precision therapeutic strategy for HCC associated with hyperglycemia.