LncRNA MALAT1 Adsorbs MiR-384 To Target And Regulate The Effect Of NFKBIA On Apoptosis Of Meningioma Cells

Objectives:Based on the results of bioinformatics prediction,in vitro experiments were performed to validate LncRNA MALAT1 as ceRNA adsorbed miR-384 to regulate the expression of its target gene NFKBIA to affect the apoptosis of meningioma cells.Methods:Meningioma-related data were downloaded from the NCBI Gene Expression Omnibus database(GEO),and the limma package in R language software was used to screen differential mRNAs and differential miRNAs in meningioma and normal meningeal tissue samples;the colorspace package,stringi package,ggplot package,DOSE package,clusterProfiler package,enrichplo package of R language 3.6.2 software were used to GO enrichment analysis of differential mRNAs;Cytoscape 3.7.2 software was used to analyze differential mRNAs and differential miRNAs with intersection;based on the review of literature,the NFKBIA and miR-384 were selected by combining the results of bioinformatics methods screening.Subsequently,we applied TargetScan online tool to predict whether miR-384 and NFKBIA may have binding sites,followed by our bioinformatic prediction of Inc RNAs that miR-384 may bind using ENCORI for RNA Interactomes online tool;Fresh meningioma tissues were obtained during neurosurgery and verified by immunohistochemistry that they were indeed meningioma tissues;subsequently,PCR was used to verify the expression of screened NFKBIA,miR-384 and LncRNA MALAT1 in meningiomas;meningioma cell lines were purchased and cultured for passaging;overexpression plasmids of NFKBIA were constructed and lentiviral packaging was performed;NFKBIA was used to the human meningioma cell lines were infected with NFKBIA overexpression lentivirus,and the fluorescence expression and cell growth status were observed by fluorescence microscopy after 24h,48h and 72h of infection and photographed to determine the optimal complex of infection(MOI)value.The experiments were divided into three groups:Normal group:human meningioma cell lines without any intervention;NC group:human meningioma cell lines cultured after empty lentivirus infection;NFKBIA-ORF group:human meningioma cell lines cultured after NFKBIA overexpression lentivirus infection.After lentivirus infection of human meningioma cells for 72 h,the expression of NFKBIA was detected by PCR,the proliferation rate of cells was detected by CCK8,and the apoptosis of cells was detected by flow cytometry,TUNEL assay and immunofluorescence staining.Results:In this study,we used bioinformatics prediction to find a total of 317 differentially expressed mRNAs,of which 17 were up-regulated and 300 were down-regulated;223 differentially expressed miRNAs,of which 15 were up-regulated and 208 were down-regulated;GO enrichment analysis of differential mRNAs mainly showed molecular functions such as structural components of cell matrix and glycosaminoglycan binding,leukocyte migration,biological processes such as response to lipopolysaccharide and response to bacterial-derived molecules,extracellular matrix containing collagen,cellular composition such as secretory granule lumen;meanwhile,combined with the results of literature review,key mRNA NFKBIA and key miRNA miR-384 were obtained to act directly in meningioma;prediction of possible binding sites for miR-384 and NFKBIA using the TargetScan online tool,the online tool ENCORI For RNA Interactomes was used to predict that miR-384 might bind to the Inc RNA MALAT1.NFKBIA,miR-384 and LncRNA MALAT1 were expressed in meningioma;PCR results showed elevated NFKBIA expression after 72h of NFKBIA overexpression lentivirus infection of meningioma;CCK8 assay found that the proliferation of meningioma cells was inhibited after 72h of NFKBIA overexpression lentivirus infection of human meningioma cells;TUNEL In order to further verify the above results,flow cytometry and immunofluorescence staining were done,and both results showed that the value-added rate of meningioma cells decreased and the apoptosis rate increased after 72h of NFKBIA overexpression lentivirus infection of human meningioma cells.Conclusions:NFKBIA overexpression significantly inhibited the proliferation of brain meningioma cells and promoted the onset of apoptosis.In addition,NFKBIA promotes apoptosis in meningioma cells probably through LncRNA MALAT1 as ceRNA adsorbing miRNA-384 and then affecting its expression to achieve a pro-apoptotic effect.