| Objective:To investigate the effects of LINC00629 on NSCLC proliferation,migration,invasion and EMT through regulation of miR-3182/MMP2 axis.Method:We detected the expression of LINC00629 in BEAS-2B and NSCLC cell lines and NSCLC tissues and paracancerous tissues by q RT-PCR.The Gene Expression Profile Interaction Analysis(GEPIA)database was used to detect the expression of LINC00629 in NSCLC tissues and paracancerous tissues and the relationship between LINC00629 expression levels and disease-free survival of NSCLC patients.In addition,after knockdown or overexpression of LINC00629 expression,the effects of LINC00629 on the proliferation,migration,invasion and EMT of NSCLC were demonstrated by CCK8 assay,wound healing assay,transwell invasion and WB assay.The distribution of LINC00629 was localized by Subcellular fractionation assay,and the application of Lnc Baseand GEO databases to predict the miRNAs that LINC00629 may bind,and to detect of the targeting relationship between LINC00629 and miR-3182 using a dual luciferase reporter assay using a dual luciferase reporter.Further CCK8 assay,Wound healing assay and transwell invasion were used to verify whether miR-3182 affected the proliferation,migration and invasion ability of LINC00629.q RT-PCR and WB assays were applied to verify the regulation of MMP2 expression by expression of LINC00629 with miR-3182.Finally,the effect of LINC00629 on the proliferation of NSCLC cells in vivo was investigated by subcutaneous tumorigenesis assay in nude mice.Result:LINC00629 is highly expressed in NSCLC tissues and cell lines.The results of cell function assays showed that knockdown of LINC00629 expression inhibited the proliferation,migration,invasion and EMT ability of NSCLC.Similarly,overexpression of LINC00629 expression promoted proliferation,migration and invasion of NSCLC cells.Subcellular fractionation assay demonstrated that LINC00629 was predominantly located in the cytoplasm,and the results of dual luciferase assay showed that the luciferase activity was reduced in the LINC00629wild-type vector with the addition of miR-3182 mimics in the co-transfection group,demonstrating LINC00629 can bind to miR-3182.Subsequently,by q RT-PCR and Western blotting,we found that overexpression of LINC00629 in two NSCLC cell lines significantly promoted the expression of MMP2 at the transcriptional and translational levels,which could be disrupted by miR-3182 mimics,and the results demonstrated that LINC00629 could inhibit the expression of miR-3182 and increased MMP2 levels.Finally,we performed subcutaneous tumorigenesis experiments in nude miceand,consistent with the in vitro results,the tumor volume and weight of sh2-LINC00629 micewere lower than those of the sh-NC group.The expression of miR-3182 was significantly increased in sh2-LINC00629 compared to the control group.The same results showed that MMP2 expression was found to be lower in the sh2-LINC00629 group.Conclusion:LINC00629 is highly expressed in NSCLC cell lines and tissues.Knockdown of LINC00629 inhibited proliferation,migration,invasion and Epithelial mesenchymal transition(EMT)in NSCLC,Simultaneous overexpression of LINC00629 promotes the proliferation,migration and invasive ability of NSCLC.Furthermore LINC00629 was mainly distributed in the cytoplasm and LINC00629 accelerated the progression of NSCLC by regulating the miR-3182/MMP2 axis.The results of this study provide a new prognostic marker and therapeutic target for NSCLC. |
