MicroRNA-3666 Suppresses Cell Growth in Head and Neck Squamous Cell Carcinoma Through Inhibition of PFKFB3-Mediated Warburg Effect

• Published in: OncoTargets and therapy 2020, Vol.13, p.9029-9041
• Main Authors: Chen, Lan, Cao, Yaxuan, Wu, Bei, Cao, Yu
• Format: Article
• Language: English
• Subjects: Acidification; Apoptosis; Bioinformatics; Cell growth; Cell proliferation; Colorectal cancer; Experiments; Gastric cancer; Gene expression; Glucose; Glycolysis; Head & neck cancer; Kinases; Lactic acid; MicroRNAs; miRNA; Original Research; Oxygen consumption; Phosphofructokinase; Polymerase chain reaction; Protein expression; Proteins; Software; Squamous cell carcinoma; Standard deviation; Tumor suppressor genes; Tumors
• ISSN: 1178-6930; 1178-6930
• DOI: 10.2147/OTT.S251992

MicroRNA-3666 (miR-3666) is aberrantly expressed and plays critical roles in numerous human tumors. However, the expression pattern, biological role, and mechanisms of action of miR-3666 in head and neck squamous cell carcinoma (HNSCC) remain unknown. Therefore, we attempted to determine the expression status and function of miR-3666 in HNSCC and to explore the underlying mechanisms in detail. In this study, quantitative real-time polymerase chain reaction was carried out to measure the expression of miR-3666 HNSCC tissues. A series of experiments, including a Cell Counting Kit-8 assay, colony formation assay, BrdU incorporation and apoptosis analysis, were applied to test whether miR-3666 affects the growth of HNSCC cells. Glucose uptake and lactate production measurements and extracellular acidification and oxygen consumption rate assays were conducted to determine the effect of miR-3666 on glycolysis. We found that miR-3666 showed a decreased expression in HNSCC tissues. Further functional studies demonstrated that miR-3666 inhibited the growth of HNSCC cells by suppressing cell proliferation and promoting apoptosis. Bioinformatics analysis and luciferase reporter assays identified phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3), a key enzyme regulating glycolysis, as a direct target of miR-3666. Through inhibition of PFKFB3, miR-3666 decreased glycolysis in HNSCC cells by reducing the production of F2,6BP. Importantly, glycolysis suppression caused by miR-3666 was found to be required for its inhibitory effect on HNSCC cell growth. Our data suggest that miR-3666 functions as a tumor suppressor by decreasing the rate of glycolysis through inhibition of PFKFB3 activity, and this miRNA may present a potential candidate for HNSCC therapy.