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

Abstract

Lan Chen,1,2 Yaxuan Cao,2 Bei Wu,3 Yu Cao1 1Department of Dermatology, Affiliated Hospital of Guiyang Medical University, Guiyang, People’s Republic of China; 2University of Health, Guizhou Medical University, Guiyang, People’s Republic of China; 3Department of Obstetrics and Gynecology, 925 Hospital of the Joint Logistics Support Force of the Chinese People’s Liberation Army, Guiyang, People’s Republic of ChinaCorrespondence: Yu CaoDepartment of Dermatology, Affiliated Hospital of Guiyang Medical Univesity, 28 Guiyi Street, Yunyan District, Guiyang, Guizhou 550004, People’s Republic of ChinaTel [phone]Fax [phone]Email [email protected] WuDepartment of Obstetrics and Gynecology, 925 Hospital of the Joint Logistics Support Force of the Chinese People’s Liberation Army, 67 Huanghe Road, Huaxi District, Guiyang, Guizhou 550009, People’s Republic of ChinaTel [phone]Fax [phone]Email [email protected]: 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.Methods: 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.Results: 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.Conclusion: 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.Keywords: microRNA-3666, head and neck squamous cell carcinoma, PFKFB3, Warburg effect

Keywords

• microrna-3666
• head and neck squamous cell carcinoma
• pfkfb3
• warburg effect.