| Aims:Epithelial-mesenchymal transition(EMT)is closely related to tumor occurrence and a series of malignant progression,micro RNA(mi RNA)can regulate EMT by regulating related genes.This study mainly observed the regulatory effect of mi R-205-5p on EMT of cisplatin-resistant nasopharyngeal carcinoma cells,and clarified its mechanism,in order to provide a theoretical and experimental basis for the clinical treatment of nasopharyngeal carcinoma.Methods:1.mi RNA microarray analysis was performed to detect the changes of mi RNAs expression in cisplatin-resistant nasopharyngeal carcinoma cells and sensitive strain cells,and mi RNAs with significant expression differences were selected and verified by q RT-PCR technology.2.Lipofectamine 2000 was used to transfect mi R-205-5p mimic and mi R-205-5p inhibitor into HNE1 cells or HNE1/DDP cells to change the expression level of mi R-205-5p and detect the regulation of mi R-205-5p on cell invasion and mignation and EMT in nasopharyngeal carcinoma cells.3.MTT assay was employed to detect the proliferation and survival of nasopharyngeal carcinoma cells;colony formation assay was performed to observe the effect of mi R-205-5p on cell colony forming ability;Annexin V/PI assay was used to detect apoptosis of nasopharyngeal carcinoma cells.4.Wound healing assay was carried out to assess cell migration ability of HNE1 cells and HNE1/DDP cells.Transwell assay was applied to detect cell migration and invasion in vitro.5.Inverted microscope was used to observe changes in cell biological morphology;Western blot and q RT-PCR were employed to detect the changes of EMT-related marker molecules E-cadherin,N-cadherin,Vimentin,MMP-2,MMP-9 and EMT-induced transcription factors Snail and Slug in the protein levels and m RNA levels.6.Target Scan and STRING analysis were applied to predict the potential target genes of mi R-205-5p,and detect the regulatory effect of mi R-205-5p on target genes.7.The related signaling pathways of mi R-205-5p regulating EMT were explored.Results:1.Compared with HNE1 cells,HNE1/DDP cells were resistant to cisplatin and the resistance index(RI)was 4.95.2.Mi RNA microarray analysis was performed to analyze the expression of mi RNAs in HNE1 and HNE1/DDP cells,and the most significant dysregulated was mi R-205-5p.3.The results of MTT assay,Annexin V/PI assay and Western blot results showed that after down-regulating the expression of mi R-205-5p in HNE1/DDP cells,the sensitivity of the cells to cisplatin was significantly enhanced.4.Transfection of mi R-205-5p inhibitor in HNE1/DDP cells and mi R-205-5p mimic in HNE1 cells to regulate the expression level of mi R-205-5p.Up-regulating the expression of mi R-205-5p could significantly enhance the proliferation,invasion and migration of HNE1 cells and induce EMT;inhibiting the expression of mi R-205-5p in HNE1/DDP cells significantly inhibited the cell proliferation,invasion and migration of HNE1/DDP cells,and reverse the EMT of HNE1/DDP cells.5.PTEN is a downstream candidate gene for mi R-205-5p.Inhibiting the expression of PTEN,the proliferation,invasion and migration ability of nasopharyngeal carcinoma cells were significantly enhanced,and the effect of mi R-205-5p inhibitor was counteracted.6.The regulation of EMT by mi R-205-5p was mediated by the PI3K/AKT signaling pathway.Blocking the PI3 K / AKT signaling pathway could effectively block the promotion of mi R-205-5p mimic on HNE1 cell invasion and migration and EMT.Conclusions:1.mi R-205-5p is significantly overexpressed in cisplatin-resistant nasopharyngeal carcinoma cells,which is closely related to the occurrence of EMT in the cells;2.Overexpression of mi R-205-5p can promote cell invasion,migration and the occurrence of EMT;on the contrary,inhibition of mi R-205-5p expression in cells can reverse the EMT process of cells and enhance the sensitivity of the HNE1/DDP cells to cisplatin;3.mi R-205-5p regulates EMT in nasopharyngeal carcinoma cells by targeting PTEN via PI3K/AKT signaling pathway;... |
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