The Changes Of Expression And Subcellular Location Of LncRNA In LPS-induced Inflammation In Human Colon Cancer Cells

Colorectal cancer is one of the most common cancer of digestive tract in the world.Morbidity and mortality of colorectal cancer in China have been on the rise because of the changing living conditions these years.Causation factor of colorectal is a highly complex because of its multi-sources.Besides some research in decades revealed that there is a strong relationship between colorectal cancer and billions of microbes in human bowel.However the molecular mechanism between CRC and gut microbesOne component of Gram-negative bacteria,lipopolysaccharide,can be recognized by TLR4 which reside in macrophase cell membrane to induce cell producing massive inflammational factors,thus could be widely used in many animal inflammation model and cell inflammation model.Lipopolysaccharide can invade into human body in the process of ulcerative colitis and raise the risk of colorectal cancer heavily.The stimulation of Lipopolysaccharide can cause inflammatory reaction of colorectal cancer cell lines.We performed whole transcriptome sequencing on the inflammation model after a series of reliable verifications in mRNA and protein level.And bioinformatic analysis was used to evaluate the inter group data quality and the expression correlation between samples.In organism,one gene plays its biological role by working with other genes or protein,which is the reason of pathway and network of gene existing.KEGG(Kyoto encyclopedia of genes and genomes)is a main database about pathway information.We used it to analysis pathway information of 250 mRNAs which are alter-regulated in our inflammation models and confirmed that TNF activation and NF ? B signal pathway is the most active signal pathways in the model.We also screened out five IncRNA Fanl,Larp7,Rrebl,Cxcll and Tnf,have the same expression changes both in our sequencing data and TCGA database.We found a lncRNA named PSMD5-AS1 in GEO database which has the same expression change in our sequencing data.We divided the RNA of LPS-stimulated cells into cytoplasmic part and nuclear part and then we performed whole transcriptome sequencing to investgate the relationship between lncRNAs' subcellular location and their function.We counted the number of cytoplasmic lncRNA and nuclear lncRNA.After analysis we found a significant rise occupation ratio of cytoplasmic lncRNA in 3h sample.This result implicate that some lncRNAs were exported to cytoplasm from nucleus.Then we performed qRT-PCR to examined the expression change of 5 lncRNAs we chose.We found that LINC00152 changed its subcellular location in SW480 which stimulated by LPS.Taken together,we performed whole transcriptome sequencing over colorectal cancer cell to reveal gene expression changes with time and space.This work can help us having a better understanding of gene expression change of inflammation model cell and give a reference to the future research of colorectal cancer triggered by inflammation.