Effects of overexpression of miR-498 on proliferation,invasion,migration,and EMT of nasopharyngeal carcinoma cells
Objective To investigate the effects of overexpression of microRNA-498(miR-498)on proliferation,invasion,migration,and epithelial-mesenchymal transition(EMT)of nasopharyngeal carcinoma cells and their mecha-nisms.Methods Human immortalized nasopharyngeal epithelial cells NP69 and human nasopharyngeal carcinoma cell lines CNE-2Z,CNE-1,HNE-1,SUNE-1 were cultured in vitro.RT-qPCR was used to detect the relative expression of miR-498 in each cell line,and nasopharyngeal carcinoma cells with relatively low miR-498 expression were selected for subsequent experiments.Nasopharyngeal cancer cells in the 3rd generation,which were in logarithmic growth phase and had good growth status,were randomly divided into the miR-498 mimics group and NC mimics group,and were transfected with miR-498 mimics and NC mimics,respectively.After 6 hours of transfection,the medium was replaced with fresh cul-ture medium and the culture continued for 24 h.Cell activity was detected using the CCK-8,colony formation ability was detected using plate cloning assay,cell invasion and migration abilities were detected using Transwell cell assay,and the expression of EMT-related epithelial cadherin(E-cadherin),Vimentin,Fibronectin,and high-mobility group protein A2(HGMA2)was detected by Western blotting.Results The relative expression levels of miR-498 in the human nasopha-ryngeal carcinoma cells CNE-1,HNE-1,CNE-2Z,and SUNE-1 were lower than that in NP69 cells(all P<0.05).In the nasopharyngeal carcinoma cells,the relative expression level of miR-498 in CNE-2Z cells was relatively low,so CNE-2Z cells were selected for subsequent experiments.After verification,the relative expression level of miR-498 in the miR-498 mimics group was higher than that in the NC mimics group(P<0.05).The proliferation activity of cells in the miR-498 mimics group at 0,24,48,and 72 h of culture was lower than that in the NC mimics group(P<0.05).The colony forma-tion,invasion and migration abilities of miR-498 mimics group were lower than those of the NC mimics group(all P<0.05).The relative expression levels of Vimentin,Fibronectin,and HMGA2 proteins in the miR-498 mimics group were lower than those in the NC mimics group,while the relative expression level of E-Cadherin protein was higher than that in the NC mimics group(all P<0.05).Conclusion The expression of miR-498 in nasopharyngeal carcinoma cells is low;overexpression of miR-498 can inhibit the proliferation,invasion,migration,and EMT of nasopharyngeal carcinoma cells,and its mechanism may be related to targeted regulation of HMGA2.
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