Atherosclerosis Related LNCRNA Expression Profile Analysis And Preliminary Screening

Objective:In this study,the whole transcriptome sequencing will be performed using high-throughput sequencing technology to screen the differentially expressed lncRNAs,and the key lncRNAs involved in the formation of atherosclerosis will be further verified by PCR in tissue and blood samples to identify and predict their related target genes,so as to provide a theoretical basis for the early diagnosis and treatment targets of atherosclerosis.Methods : 1.Acquisition of 5 cases of atherosclerotic tissues and 5 normal human vascular endometrial tissues,and RNA(mRNA,LNCRNA and miRNA)gene expression spectrometry.1.Acquisition of atherosclerotic plaque tissues and normal human vascular intima tissues,and sequencing of RNA(mRNA,lncRNA and miRNA)gene expression profiles.2.The differentially expressed levels of the three RNAs(lncRNA,mRNA and miRNA)were obtained by transcriptome sequencing,and the differentially expressed genes were analyzed by KEGG and GO function enrichment.3.The top 10 significantly up-regulated lncRNAs and significantly down-regulated lncRNAs were screened to analyze the co-expression network of lncRNA-mRNA to construct the ceRNA network and predict the target genes.4.PCR was used to screen and verify the target lncRNAs with significantly different expressions in tissue samples and blood samples.Results:1.Transcriptome gene sequencing results showed that 89 miRNAs were up-regulated and 42 miRNAs were down-regulated.4677 mRNA were up-regulated and 2347 mRNA were down-regulated.707 lncRNAs were up-regulated and 250 lncRNAs were down-regulated.2.The results of differential lncrna-related target genes KEGG and GO analysis showed that immune and inflammatory reactions,T-cytokine pathway and so on may play an important role in arteriosclerosis formation.3.By screening 20 target lncRNA correlation analysis,it was predicted that 2 miRNA(hsa-let-7c-5p,hsa-miR-3194-3p)and 9 mRNA(PPP1R15B,THBS1,BTN3A2,GOLGA7,MLEC,RAB11FIP1,SVOP,TBPL1,TTC9)might be closely related to atherosclerosis.4.At the level of lncRNA,the preliminary screening results of PCR verification of tissue samples showed that four lncRNA(LINC01094,HIF1A-AS2,HCP5,ENSG00000257027.1)were significantly up-regulated in arteriosclerosis tissues.The five lncrnas(acta2-as1,MAGI2-AS3,LINC01278,PRKG1-AS1,LOC102724804)were significantly down-regulated in arteriosclerosis tissues.PCR verification results of blood samples showed that LINC01094 was significantly up-regulated in patients with carotid plaque stenosis.ACTA2-AS1 and ENSG00000257027.1 were significantly down-regulated in patients with carotid plaque stenosis.Conclusion : 1.Immune and inflammatory reactions,T-cytokine pathway and so on play an important role in the formation of arteriosclerosis.2.RNA correlation analysis of target LNC showed miRNA(hsa-let-7c-5p,hsa-miR-3194-3p)and mRNA(PPP1R15B,THBS1,BTN3A2,GOLGA7,MLEC,RAB11FIP1,SVOP,TBPL1,TTC9)3.PCR results showed that up-regulation of LINC01094 and down-regulation of ACTA2-AS1 and ENSG00000257027.1 were of great significance as diagnostic markers and treatment of atherosclerosis.