Effects of miR-28-5p on the proliferation of oral squamous cell carcinoma cells by targeting DOK4
Objective To explore the regulatory effect of microRNA-28-5p(miRNA-28-5p)on the proliferation of oral squamous cell carcinoma(OSCC)cells by targeting tyrosine kinase/docking protein 4(DOK4).Methods The Cancer Genome Atlas(TCGA)oral carcinoma database was downloaded to analyze the relationship between miR-28-5p,DOK4 and clinical phenotypes of OSCC.The cells were transfected with miR-28-5p mimics,miR-28-5p inhibitor and negative control(NC),pcDNA3.1-DOK4,empty vector(Vector)and DOK4 interference sequence(siDOK4)to detect cell proliferation,antioxidant capacity and oxygen species(ROS)level.The targeted relationship between miR-28-5p and DOK4 was verified by dual luciferase reporter assay.The effects of DOK4 overexpression on growth of xenografted tumors in OSCC nude mice were observed.Results Compared with para-carcinoma tissues,miR-28-5p was up-regulated,while DOK4 mRNA was down-regulated in OSCC tissues(P<0.05).The level of miR-28-5p in patients with clinical staging of Ⅳ,stage M1 and grade 3-4 was higher than that of clinical staging Ⅰ-Ⅲ,stage M0 and grade 1-2,and the level of DOK4 mRNA in patients with clinical staging of Ⅳ,stage N1 and grade 3-4 was lower than that of clinical staging Ⅰ-Ⅲ,stage N0 and grade 1-2(P<0.05).The progression-free survival and overall survival in high-expression DOK4 group were higher than those in low-expression group(P<0.05).Compared with miR-NC group,level of miR-28-5p,cell activity and colony count in miR-28-5p inhibitor group were decreased(P<0.05).Compared with miR-NC group,expressions of DOK4 mRNA and protein were decreased in miR-28-5p mimics group(P<0.05).Compared with Vector group,expressions of DOK4 mRNA and protein in cells and xenografted tumors increased in DOK4 group,while cell activity,colony count,volume and mass of tumors decreased(P<0.05).Compared with miR-28-5p inhibitor group,expressions of DOK4 mRNA and protein decreased in miR-28-5p inhibitor+siDOK4 group,activity of cell proliferation,colony count,nicotinamide adenine dinucleotide phosphate-reduced/nicotinamide adenine dinucleotide phosphate(NADPH/NADP+)and glutathione/oxidized glutathione(GSH/GSSG)increased,and ROS level decreased(P<0.05).Conclusion The expression of miR-28-5p is up-regulated in OSCC,which can promote the proliferation of OSCC cells by inhibiting DOK4 expression and reducing ROS level.
Oral squamous cell carcinomaProliferationMicroRNA-28-5pDownstream of tyrosine kinase/docking protein 4Reactive oxygen specie
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