Biological Function And Regulation Mechanism Of Linc00619 In Human Lung Epithelial Cells

Objective: To explore the role of linc00619 in human lung epithelial cells and its regulatory mechanism,thus providing a potential target for the treatment of allergic asthma.Methods: Bioinformatics methods were used to perform data mining on the public dataset GSE8190(dust mite-induced human airway epithelial cell expression profile dataset);CY3-labeled probes were used for RNA-FISH method on linc00619 in human lung epithelial cells(BEAS-2B)was subjected to subcellular localization;the dgRNA lentiviral vector of linc00619 was constructed by CRISPR/Cas9 method and confirmed by sequencing.The BEAC-2B cells were transiently transfected with the dgRNA lentiviral vector of linc00619,and the stably transformed strain was obtained after puromycin selection.The effects of linc00619 knockdown on cell proliferation and migration of BEAS-2B cells were analyzed.Western blot analysis of proliferation-related signal molecules such as AKT,MEK and ERK;lncRNA microarray was used to detect differential expression of lncRNAs and mRNAs after linc00619 knockdown.mRNAs were analyzed by GO and KEGG enrichment;cis and cisregulated target genes of linc00619 were predicted and analyzed by GO and KEGG enrichment.Results: Linc00619 is highly expressed in HDM-induced epithelial cells(>4 fold);linc00619 is localized in cytoplasm;lincRNA lentiviral vector of linc00619 is transiently transfected into BEAS-2B cells,and puromycin is screened to obtain stable transgenic plants,PCR detection and sequencing;The results showed that the dgRNA mutant strain of linc00619 was successfully obtained.Cell proliferation assay and Transwell assay showed that linc00619 knockdown promoted cell proliferation and migration;WB assay showed that linc00619 knockdown inhibited p-AKT/AKT,p-MEK/MEK expression and promoted pERK /ERK expression.GO and KEGG enrichment analysis of significantly differentially expressed mRNAs after knockdown of the linc00619 knockdown assay showed that linc00619 participates in the biological function of BEAS-2B through multiple biological processes and multiple signaling pathways.This phenomenon is also present in the cisc and trans-regulated target genes of linc00619.Conclusion:1.Linc00619 is located in the cytoplasm;2.Linc00619 inhibits the proliferation and migration of lung epithelial cells;3.Linc00619 participates in the development of allergic asthma through multiple biological processes and multiple signaling pathways;...