Analysis and functional study on lncRNA expression profile in primary biliary cirrhosis patients

Abstract

Objective Analyzing the lncRNA expression profile in peripheral blood mononuclear cells (PBMC) of primary biliary cirrhosis (PBC) patients to provide new ideas for the pathogenesis, clinical diagnosis and treatment of PBC. Methods Collected peripheral blood from 30 PBC patients and 30 healthy volunteers, then separated PBMC. Four cases from each group were selected for long-noncoding RNA (lncRNA) expression microarray detection. Reverse transcription-PCR technology in a larger sample size was used to verify the microarray results. Bioinformatic analysis such as Cis- / Trans- target genes Gene ontology (GO) and pathway analysis, co-expression networks were conducted in order to provide a theoretical basis in the pathogenesis of PBC. Transfecting small interfering RNAs(siRNAs) for linc-pbc to see changes in the expression of nuclear receptor 4A group 3(NR4A3) and forkhead boxP3(FOXP3) and cell apoptosis in transfected PBMC. Results Compared to the healthy group, 749 lncRNA and 230 coding messenger RNA(mRNA) genes were abnormally expressed. Interestingly, NR4A3 gene was down-regulated by 78%. While linc-pbc, which was about 20 000 bp downstream of NR4A3 gene, increased 2.56-fold. Then design siRNA for linc-pbc. After transfection, mRNA and protein levels of NR4A3 and FOXP3 were up-regulated. Conclusions By recruiting PRC2 complex, linc-pbc may increased the methylation level of NR4A3 gene promoter region, thus decreasing the expression of NR4A3 in PBC patients and reduced NR4A3 futher downregulated the expression of FOXP3 and reduced the amounts of immunosuppressive Treg cells in peripheral blood and liver tissue, breaking the equilibrium state of immune tolerance, and promoted the occurrence and development of the disease.(Chin J Lab Med, 2018, 41: 374-379) Key words: Liver cirrhosis, biliary; RNA, long noncoding; DNA-binding proteins