Promoting effects of miR-888-5p on proliferation and invasion of NSCLC cells and their mechanism
Objective To observe the effects of miR-888-5p on the proliferation and invasion abilities of non-small-cell lung cancer(NSCLC)cells,and to explore the possible mechanism.Methods Real-time fluorescence quantitative PCR was used to detect the relative expression levels of miR-888-5p in NSCLC cell lines(A549,NCI-H1299,PC-9,LLC,and NCI-H1650)and human normal lung epithelial cell line(BEAS-2B).A549 cells with the most significant changes in the miR-888-5p relative expression were selected for subsequent experiments.A549 cells were divided into the miR-888-5p mimics group,miR-888-5p inhibitor group,and scramble group,which were transfected with miR-888-5p mimics,miR-888-5p inhibitor,and scramble sequences,respectively.Real-time fluorescence quantitative PCR was used to detect the relative expression level of miR-888-5p in cells.MTT assay was used to detect the cell proliferation abilities at 0,24,48,and 72 h of culture.Transwell chamber invasion assay was used to detect the cell invasion ability(expressed as the number of invading cells).StarBase online prediction website predicted that the target gene of miR-888-5p was Tob1,which was verified through luciferase assay.Western blotting was used to detect the relative expression levels of Tob1,E-cadherin,vimentin proteins,p-JAK2/JAK2,and p-STAT3/STAT3 in cells.Results The relative expression levels of miR-888-5p in the miR-888-5p mimics group,miR-888-5p inhibitor group,and scramble group were 48.9±1.78,1.06±0.24,and 8.23±0.90,respectively,with statistically significant difference between groups(all P<0.05).Over the time of culture,the proliferation abilities of the three groups showed an increasing trend.At the same time point,the cell proliferation abilities and the invasive cells decreased in the miR-888-5p mimics group,scramble group,and miR-888-5p inhibitor group in turn,with statistically significant difference between groups(all P<0.05).The relative expres-sion levels of Tob1 and E-cadherin proteins in the miR-888-5p mimics group,scramble group,and miR-888-5p inhibitor group increased sequentially,while the relative expression levels of vimentin protein and p-JAK2/JAK2,p-STAT3/STAT3 decreased sequentially(all P<0.05).Conclusion MiR-888-5p can enhance the proliferation and invasion abilities of NSCLC cells,and their mechanism may be related to the activation of JAK2/STAT signaling pathway and targeted down-regulation of Tob1 expression,thereby promoting the epithelial-mesenchymal transition.
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