The Regulatory And Prognostic Roles Of Linc00673/miR-150-5p/ZEB1 Axis In Non-small Cell Lung Cancer

Background and aims:Lung cancer has the highest incidence and is the leading cause of cancer-related deaths among all cancers worldwide.Metastasis is closely related to the poor prognosis of lung cancer patients,in which epithelial-mesenchymal transition(EMT)is a key event.Previous studies reported that long noncoding RNAs(lnc RNAs)play an important regulatory role in a variety of tumors,including non-small cell lung cancer(NSCLC).The purpose of this study was to explore the regulatory functions and molecular mechanisms of lnc RNA linc00673 on cell proliferation,migration,invasion and EMT in NSCLC as well as its prognostic significance.Methods:CCK-8 and EdU assay were used to detect cell proliferation ability;wound scratch assay and Transwell invasion assay were used to assess cell migration ability and invasion ability,respectively;real-time quantitative PCR was adopted to determine the expression levels of RNA molecules and transcription levels of EMT biomarkers;immunoblotting and immunofluorescence were conducted to examine protein expression levels of EMT biomarkers.Dual-luciferase reporter assays were performed to assess the interaction between miRNA and its potential targets.Mouse xenograft model was used to determine the regulatory role of linc00673 on tumor formation in vivo.Bioinformatics and evidence-based medicine methods were adopted to analyze the prognostic significance of linc00673/miR-150-5p/ZEB1 axis in NSCLC.Results:Linc00673 promoted cell proliferation,migration,invasion and EMT in NSCLC.Knockdown of linc00673 significantly reversed the EMT induced by TGF-?,and inhibited the tumor formation in vivo as well.Next,we found direct interaction between linc00673 and miR-150-5p.Overexpression of miR-150-5p suppressed the expression of linc00673,while inhibition of miR-150-5p significantly uplifted linc00673 expression level.Dual-luciferase reporter assay supported such results too.Moreover,miR-150-5p could also target the 3' untranslated regions of ZEB1,a key transcription factor in EMT,and thus lead to ZEB1 inhibition.Therefore,linc00673 promoted EMT in NSCLC through inhibition of miR-150-5p and abrogating its suppressing effect on ZEB1.In addition,high expression of linc00673 indicated poor prognosis of NSCLC patients,while high expression of miR-150-5p was associated with better outcomes.In the analysis of the prognostic significance of ZEB family members,5 cohort studies were included and the results were pooled.We found that high expression level of ZEB1 was also significantly associated with the poor prognosis of NSCLC patients.Conclusions:Linc00673 promotes cell proliferation,migration,invasion and EMT in NSCLC through sponging miR-150-5p as ce RNA and indirectly upregulating the expression level of ZEB1.High expression levels of Linc00673 and ZEB1 indicate poor prognosis of NSCLC patients,while high expression level of miR-150-5p is associated with better outcomes.In summary,linc00673/miR-150-5p/ZEB1 axis plays a key regulatory role and is of great prognostic significance in NSCLC.