Study On The Regulatory Effect And Mechanism Of MiR-374a/b-5p On Invasion And Migration Of Head And Neck Squamous Cell Carcinoma Cells

Objective:Head and neck squamous cell carcinoma is one of the most common malignant tumors in human beings,and its incidence ranks sixth among systemic tumors and the highest number of primary sites and pathological types among all tumors in the human system that seriously brings risk to human health.Predisposition and lymph node metastasis are important factors leading to unpleasant outcomes of head and neck squamous cell carcinoma.micro RNA(miRNA)occupies significant roles in the regulation of tumorigenesis and development,but the expression pattern,role and mechanism action of miRNA in head and neck squamous cell carcinoma still need to be further studied.We aim to evaluate the expression and biological functions of miR-374a-5p and miR-374b-5p in head and neck squamous cell carcinoma through combining bioinformatics methods and in vitro and in vivo experimental techniques,and their mechanism of action is further revealed.This study will provide a new perspective for understanding the mechanism of head and neck squamous cell carcinoma oncogenesis and development.Methods:The expression levels of miR-374a-5p and miR-374b-5p in head and neck squamous cell carcinoma were analyzed using TCGA database head and neck squamous cell carcinoma transcriptome sequencing data.CCK-8 kit detected the cell viability,Transwell assay performed the cell invasion and migration ability,the glycolytic ability of cells was analyzed by seahorse energy detector,and the effect of miR-374a-5p and miR-374b-5p on tumorigenic ability in vivo was evaluated via nude mouse xenograft model.Real-time quantitative PCR(RT-q PCR),western blotting,dual-luciferase reporter analysis to study the regulatory effect and mechanism of miR-374a-5p and miR-374b-5p on the downstream target gene DEPDC1,and statistical analysis of DEPDC1 in head and neck squamous cell carcinoma expression and clinical significance.Results:TCGA data analysis found that the levels of miR-374a-5p and miR-374b-5p,contrast to control tissues,were significantly decreased in head and neck squamous cell carcinoma tissues.The results of cell function experiments showed that overexpression of miR-374a-5p or miR-374b-5p inhibited the proliferation,invasion and migration of head and neck squamous cell carcinoma cells,and resulted in a decrease in the level of cellular glycolysis.Nude mouse xenograft experiments showed that overexpression of miR-374a-5p or miR-374b-5p could inhibit the growth of head and neck squamous cell carcinoma xenografts.Mechanistic studies revealed that miR-374a-5p and miR-374b-5p can directly bind to the 3’UTR DEPDC1 gene up-regulated in head and neck squamous cell carcinoma respectively,thereby downregulating the expression level of DEPDC1.Further studies confirmed that DEPDC1 promoted the proliferation,migration,invasion and glycolytic metabolism of head and neck squamous cell carcinoma,while miR-374a-5p and miR-374b-5p inhibited the proliferation,migration,invasion capacity of head and neck squamous cell carcinoma by targeting down-regulation of DEPDC1 expression level.Conclusions:miR-374a-5p and miR-374b-5p act as tumor suppressor genes in head and neck squamous cell carcinoma,and inhibit cell proliferation,migration,and invasion by targeting down-regulation of DEPDC1 expression.The results of this study are expected to provide potential markers for the diagnosis and prognosis of head and neck tumors,and new potential molecular targets for the treatment of head and neck squamous cell carcinoma targeting glycolytic metabolism.