Functional Analysis And Its Effect On Migration Of LncRNA AL137782.1 In Lung Epithelial Cells

Background:Long non-coding RNA(Long non-coding RNA,LncRNA)is a type of RNA molecule larger than 200 nt.Many studies have shown that LncRNA can regulate the expression of various genes at the epigenetic,transcriptional and post-transcriptional levels,thus participating in the regulation of various biological processes.As the main cells of lung barrier and gas exchange,the normal operation of their functions plays a vital role in the homeostasis of the lung microenvironment.Recently,novel LncRNA have been discovered with the wide application of gene microarray and second-generation highthroughput sequencing technology.The regulation of new LncRNA in lung related diseases has become one of the research hotspots.The regulation of novel LncRNA in lung related diseases is one of the research hotspots.We previously found that LncRNA AL13 7782.1 was differentially expressed in injured lung epithelial cells,and the expression of cell migration related gene LIM domain only 7(LIM domain only 7,LMO7)is highly correlated with it.However,the function of lncRNA AL 13 7782.1 in lung epithelial cells and the mechanism of regulating cell migration have not been clearly reported.Objective:This study explores the expression,location and function of LncRNA AL 13 7782.1 in lung epithelial cells,and preliminarily explores the migration mechanism of lung epithelial cells by LMO7.Hopefully,these results could provide evidences for regulation and mechanism of this unknown LncRN A AL137782.1 in cell migration of lung epithelial cells.Methods:In this experiment,we chose two human lung epithelial cell lines,A549 and BEAS-2B.Some techniques such as RT-qPCR,RACE,FISH were used to explore the expression,location and biological characteristics of LncRNAAL137782.1 in lung epithelial cells.The potential function and cell migration related gene of LncRNA AL 13 7782.1 in human lung epithelial cells were explored by whole transcriptome sequencing and gene co-expression network.LncRNA AL 137782.1 overexpressed and knocked down via adenovirus vector and siRNA,respectively.The expression of cell migration related gene was detected by RT-qPCR,western blot,and the migration of lung epithelial cells were detected by cell scratch,so as to explore the molecular mechanism of LncRNA AL137782.1 affecting the migration of lung epithelial cells.Results:(1)The prediction results of bioinformatics show that LncRNA AL137782.1 cannot encode protein;the cDNA end rapid amplification results show the 5'end of LncRNA AL 13 7782.1 was 273 nt.(2)LncRNA AL 13 7782.1 was distributed in the nucleus and cytoplasm of lung epithelial cells,and overexpressed LncRNA AL 137782.1 was upregulated in the nucleus and cytoplasm of lung epithelial cells.(3)LncRNA AL137782.1 had no significant effect on proliferation and cell viability of lung epithelial cells,but transcriptomics data show that the overexpression of lncRNA AL137782.1 significantly up-regulated or down-regulated key molecules of biological processes such as adhesion junctions,cell migration and immune response in lung epithelial cells.(4)Overexpression of LncRNA AL 137782.1 in lung epithelial cells,the expression of LM07 were significantly up-regulated;on the contrary,knocking down LncRNA AL 137782.1,the expression of LMO7 were significantly down-regulated.(5)The migration of lung epithelial cells were significantly increased while overexpressed LncRNA AL13 7782.1;on the contrary,the migration of lung epithelial cells was significantly slowed down after LncRNA AL137782.1 was knocked down.But overexpressed LncRNA AL137782.1 and knocked down LMO7,the migration of lung epithelial cells were slowed down significantly.Conclusion:LncRNA AL137782.1 was expressed in the nucleus and cytoplasm of lung epithelial cells,indicating that it is important in the transcriptional regulation and post-transcriptional regulation of lung epithelial cells;LncRNA AL 137782.1 cannot regulated the proliferation of lung epithelial cells,but involved the biological processes of adhesion junction,cell migration and immune response;LncRNA AL 13 7782.1 promoted the migration of lung epithelial cells by positively regulated the expression of LMO7.