Identifieation And Expression Profiling Of Non Coding RNAs In Systemic Lupus Erythematosus Patients

Object: To further understand the pathogenesis of systemic lupus erythematosus (SLE) andclarify the relationship between systemic lupus erythematosus SLE and ncRNA using themethods of Solexa sequeneing and lncRNA microarray.Methods: Peripheral blood mononuclear cells (PBMC) from20SLE patients and20healthcontrols were collected for smll RNA extraction. Then,two groups of the small RNA librarieswere constructed respectively. Massively parallel sequencing was used to screen thedifferentially expressed miRNAs, and miRNAs expression profile of SLE was established.PBMC were isolated from blood samples of40SLE patients,20rheumatoid arthritis (RA)patients and20healthy controls. According to SLE disease activity index, SLE patients aredivied into the stable group (SW0to4points) and activities of the group (SH≥10points),20cases each Group.Total RNA was extracted were used for lncRNA array using microarray kit ofArraystar. Query the ncRNAs gene locus and biological function and other information byTargetScanHuman Custom,UCSC database, Refseq database, KEGG database with literaturefrom Pubmed.24ncRNAs were validated through Realtime-PCR.Results:We found275miRNAs differential expressed in PBMC of SLE Patients compared tothe controls. And there were153up-regulated miRNAs and122down-regulated miRNAs. Wealso found61novel miRNAs differential expressed, which18up-regulated miRNAs and43down-regulated miRNAs in SLE. In the results of microarray, there are4296lncRNAsdifferential expression of which1075up-regulated and3221down-regulated in SH groupcompared to the controls; There are3677lncRNAs differential expression of which941up-regulated and2736down-regulated in SW group compared to the controls.Compared to theSW group, there are1398lncRNAs differential expression of which654up-regulated and757down-regulated in SH group. These novel miRNAs and lncRNAs can regulate conserved targetgenes which related to SLE. Bioinformatics analyses revealed that its target genes were involvedin the gene expression, regulation of lymphocyte differentiation, DNA methylation, andsignaling pathway related to SLE.24ncRNAs were Validated by qRT-PCR, and the results wereconformable to the results of Solexa and Microarray.Conclusions:We performed the genome-wide analysis of ncRNA expression in SLE patientsusing the methods of small RNA sequeneing and lncRNA microarray. The results suggested thatthere1s a significant difference between the expression pattern of ncRNAs in SLE and healthycontrols. The resuts of this study can not only help us ferther reveal the mechanism of ncRNAs regulating pathogenesis, but also provide potential molecular biomarkers for clinieal diagnosis ofSLE.