MicroRNA-206 suppresses hypoxia-inducible factor-1α/PFKFB3-mediated glycolysis to inhibit recurrence and metastasis of hepatocellular carcinoma after incomplete radiofrequency ablation

Abstract

BACKGROUND

Hepatocellular carcinoma (HCC) remains one of the leading causes of cancer-related deaths, with high recurrence and metastasis rates after treatment. Incomplete radiofrequency ablation (iRFA) leaves residual tumor tissue that creates a hypoxic microenvironment that favors tumor progression. Emerging evidence suggests that microRNA-206 (miR-206) may act as a tumor suppressor by regulating hypoxia-inducible factor-1α (HIF-1α) and its downstream glycolytic target 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3).

AIM

To investigate whether miR-206 regulates the HIF-1α/PFKFB3/glycolysis axis in the recurrence and metastasis of HCC following iRFA.

METHODS

A clinical study was conducted in 45 patients with HCC undergoing RFA, comparing serum miR-206, HIF-1α, glucose, and pyruvate between complete (n = 35) and incomplete ablation (n = 10) groups. In vitro, tumor-derived endothelial cells (Td-ECs) exposed to sublethal thermal stress were evaluated for proliferation, migration, invasion, tube formation, and glycolysis after miR-206 mimic transfection or PFKFB3 knockdown. Dual-luciferase assays confirmed direct targeting of HIF-1α by miR-206. In vivo, a rabbit VX2 liver tumor model was used to compare angiogenesis, glycolysis, and molecular expression after complete or incomplete ablation.

RESULTS

Clinically, iRFA was associated with increased HIF-1α and pyruvate, decreased glucose, and altered miR-206 levels compared with complete ablation. In Td-ECs, thermal stimulation enhanced proliferation, migration, glycolysis, and HIF-1α/PFKFB3 expression, while miR-206 overexpression significantly attenuated these effects. Dual-luciferase assays confirmed that miR-206 directly binds the 3′UTR of HIF-1α. In animal models, incomplete ablation increased microvessel density, α-SMA, HIF-1α, and PFKFB3, while miR-206 expression was reduced.

CONCLUSION

miR-206 suppresses HIF-1α-driven PFKFB3-mediated glycolysis, thereby limiting angiogenesis, cell migration, and recurrence after iRFA. These findings suggest that miR-206 is a potential therapeutic target to reduce HCC recurrence and metastasis following ablation, although larger cohorts and in vivo rescue studies are warranted.

Keywords: MicroRNA-206; Hypoxia-inducible factor-1α; 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3; Glycolysis; Incomplete radiofrequency ablation; Hepatocellular carcinoma; Tumor recurrence; Angiogenesis

Core Tip: Incomplete radiofrequency ablation (iRFA) creates a hypoxic microenvironment that drives recurrence of hepatocellular carcinoma. This study reveals that microRNA-206 (miR-206) directly targets hypoxia-inducible factor-1α and suppresses 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3-mediated glycolysis, thereby reducing angiogenesis, migration, and metastasis after iRFA. Using patient samples, endothelial cell models, and a rabbit VX2 tumor model, we demonstrate that restoring miR-206 expression counteracts post-ablation metabolic reprogramming. These findings highlight miR-206 as a promising therapeutic target to prevent recurrence and improve long-term outcomes after local ablation therapy in hepatocellular carcinoma.