The Expression Of Long Noncoding RNAs In Lada And T2DM

Objective:To show the LncRNAs expressed in latent autoimmune diabetes in adults(LADA)and type 2 diabetes mellitus(T2DM)and predict the LncRNA target genes to derive their expression profiles for the diagnosis of T2DM and LADA and their differential diagnosis,providing a theoretical and data basis for diabetes gene target therapy.Methods:Twelve venous blood samples were collected from LADA patients(LADA group,n=4),T2DM patients(T2DM group,n=4)and normal glucose tolerance subjects(NG group,n=4)to obtain totalRNAs,using high-throughput sequencing to detect and select LncRNA with significant differential expression to construct LncRNA gene expression profiles.The adjacent site was selected as the LncRNA target by cis method,and the LncRNA function was inferred by GO and KEGG pathway analysis.Results:1、We found that the expression of LncRNA was significantly different in the data comparison of the three groups,and distributed on all human chromosomes,including sex chromosomes.2 、 Compared to normal glucose tolerance subjects,68,763 versus 28,523 LncRNAs were significantly upregulated and significantly downregulated,in T2DM patients.We predicted LncRNA target by cis method,with Fold Change >=2.0,pvalue <= 0.05,FPKM >= 0.1 as screening criteria,A total of 30 target genes with statistical significance were obtained,10 versus 20 targets were significantly upregulated and significantly downregulated.3、For LADA patients,68,748 versus 28,538 LncRNAs were significantly upregulated and significantly downregulated,relative to normal glucose tolerance subjects.We predicted LncRNA target by cis method,with Fold Change >=2.0,pvalue <= 0.05,FPKM >= 0.1 as screening criteria,A total of 18 target genes with statistical significance were obtained,6 versus 12 targets were significantly upregulated and significantly downregulated.4 、 Compared to T2DM patients,74,207 versus 23,079 LncRNAs were significantly upregulated and significantly downregulated,in LADA patients.We predicted LncRNA target by cis method,with Fold Change >= 2.0,p-value <= 0.05,FPKM >= 0.1 as screening criteria,A total of 6 target genes with statistical significance were obtained,3 versus 3 targets were significantly upregulated and significantly downregulated.5、The three LncRNAs with the largest Fold Change were selected to process the quantitative real-time polymerase chain reaction in three sets of comparative data respectively,such as LncRNA ENST00000424044-FLVCR1 、 LncRNA ENST00000432511-BTG2、LncRNA ENST00000602845-NCBP2、LncRNA ENST00000364558-HECTD4、LncRNA ENST00000608916-HCCS、LncRNA ENST00000565382-MBTPS1、LncRNA ENST00000499762-A2M、LncRNA ENST00000425189-DBH、LncRNA ENST00000436373-AP001171.1,and the results were consistent with the results of high throughput sequencing,LncRNAs target genes involved in the pathogenesis of diabetes are predicted by GO(Geneonology),KEGG pathway analysis.Conclusions:1 、 In the three sets of data among LADA/NG 、 T2DM/NG and LADA/T2DM,We found that LncRNAs had significantly different expressed in LADA、T2DM patients and normal groups.2、In the three sets of data among LADA/NG、T2DM/NG and LADA/T2DM,we acquired the cellular components、biological processes and molecular functions through GO analysis,and genetic regulatory pathways through KEGG pathway analysis of each LncRNA to speculate their mechanism in the pathogenesis of diabetes.3 、 In the three sets of data among LADA/NG 、 T2DM/NG and LADA/T2DM,we predicted nine LncRNAs targeting genes,providing gene expression profiles for diagnosis and differential diagnosis of LADA and T2DM,and providing theoretical and data basis of gene target therapy for diabetes.