Long Non-coding RNA SLCO4A1-AS1 Drives The Progression Of Non-small-cell Lung Cancer By Modulating MiR-223-3p/IKKα/NF-κB Signaling

Background:Globally,lung cancer is known as a major cause of cancer-associated death and non-small-cell lung cancer(NSCLC)accounts for majority of all cases.Growing evidence has emerged that long non-coding RNAs(LncRNAs)act as vital regulatory molecules in various malignancies.Nevertheless,the function of SLC04A1 antisense RNA 1(SLC04A1-AS1)in NSCLC is vague.It could be used as biomarkers for early diagnosis and prognosis assessment of non-small cell lung cancer,as well as become the therapeutic targets.Objective:This study aims to investigate the biological functions and possible regulatory mechanisms of lncRNA SLCO4A1-AS1 in NSCLC,so as to find new targets for the treatment of NSCLC or its iconic diagnostics,and to find new development directions for the clinical treatment of NSCLC.Methods:First,we applied qRT-PCR technology to analyze the expression level of lncRNA SLCO4A1-AS1 in NSCLC tissues and cells.Then,we performed loss-of-function detection in A549 and H1299 cells with higher expression of SLCO4A1-AS1 through CCK8 and Transwell experiments.The cell proliferation,migration,invasion of lncRNA SLCO4A1-AS1 in NSCLC were studied,and then the apoptosis of the cells and the potential level of the mitochondrial membrane in the cell were tested with the help of flow cytometry and JC-1 method;after that,we used LiCl,Jagged 1 and CD40L to activate Wnt/β-catenin,Notch and NF-κB signaling pathways,respectively,and look for the related signaling pathways involved in SLCO4A1-AS1.Then we use biological information to predict potential miRNAs that may bind to SLCO4A1-AS1 or IKKα.Afterwards,RT-PCR was used to verify the correlation between SLCO4A1-AS1,IKKa,and miR-223-3p.Next,through RIP experiments we verify whether SLCO4A1-AS1 regulates IKKα through the sponge effect on miR-223-3p.Furthermore,by transfecting miR-223-3p plasmid and siRNA targeting miR-223-3p mRNA in A549 and H1299 cells,does SLCO4A1-AS1 increase the level of IKKa by isolating miR-233-3p,thereby activating NF-κB signaling pathway in NSCLC?Finally,we use rescued experiments to explore whether SLCO4A1-AS1 promotes the progression of NSCLC by regulating IKKa in A549 cells.Results:qRT-PCR showed that SLCO4A1-AS1 levels were up-regulated in NSCLC.Functional tests show that inhibiting the expression of SLCO4A1-AS 1 can effectively reduce the proliferation,migration and invasion of NSCLC cells,and promote NSCLC cell apoptosis.In addition,we verified that SLCO4A1-AS1 activates the NF-κB pathway in NSCLC,and the NF-κB pathway related gene IKKa has an enhanced level in NSCLC tissues and cells.Importantly,miR-223-3p binds to SLCO4A1-AS1 and IKKa.In addition,SLCO4A1-AS1 competitively binds to miR-223-3p to increase the expression of IKKa,thereby activating the NF-κB signaling pathway.In conclusion,SLCO4A1-AS1 activates the NF-κB signaling pathway through sponge miR-223-3p to enhance the expression of IKKα,thereby driving the progression of NSCLC.Conclusions:SLCO4A1-AS1 droves NSCLC progression by activating NF-κB signaling pathway via sponging miR-223-3p to enhance IKKa expression.Thus,SLCO4A1AS 1 might be a promising biomarker for NSCLC treatment.