Bioinformatic analysis of the hsa-miR-1908 upstream promoter region / 临床儿科杂志

Objective To predict the functions of hsa-miR-1908 promoter using various bioinformatic tools, and to provide clues for further study on transcriptional regulation mechanism of miR-1908 in human adipocytes. Methods The promoter se-quence of miR-1908 was obtained from Ensemble, and then the CpG islands and transcription factor binding sites were pre-dicted by a variety of online bioinformatic tools. Results The length of the miR-1908 promoter sequence was 1 458 bp. The CpG islands, which inhibited the transcription of miR-1908, were located at (438-756) bp, (836-937) bp and (979-1374) bp. Meanwhile, 15 transcription factor binding sites were found in the promoter sequence of miR-1908. Conclusions miRNA up-stream promoter related bioinformatics can not only improve the efficiency of microRNA promoter research, but also provide further important information on transcriptional regulation of miR-1908.

hsa-miR-1908 target genes prediction and bioinformatics analysis / 临床儿科杂志

Objective To predict the biological process and signaling pathways in which hsa-miR-1908 might be in-volved by a series of bioinformatics analysis, so as to lay foundations and provide theoretical basis for the further studies of hsa-miR-1908 biological function in human preadipocytes. Methods The sequence of hsa-miR-1908 was acquired from miR-Base database, and target genes of hsa-miR-1908 were predicted by miRanda, and then the intersection of the results and the results of gene-chip as gene set were further analyzed by gene ontology and pathway enrichment. Results The hsa-miR-1908 had some conserved property among different species. The functions of the target genes were enriched in Wnt receptor signal-ing pathway through beta-catenin, cell cycle, cell apeptosis and other biological processes. The GnRH signaling, MAPK sig-naling, insulin signaling, cell cycle signal transduction pathways and signaling pathway in pancreatic cancer were signiifcantly enriched. Conclusions The target genes set of hsa-miR-1908 were enriched in multiple biological process which are related with the obesity. This study provides guidance for the further study in human preadipocytes.

The effect of the resistin-binding peptide (RBP) on the 3T3-L1 adipocyte differentiation,lipid metabolism and GLUT-4 gene expression / 中国糖尿病杂志

Objective To investigate the effect of RBP on 3T3-L1 adipocyte differentiation,lipid metabolism and glucose transporter 4(GLUT-4)gene expression.Methods We constructed an expression vector for rat resistin gene and transfected it into 3T3-L1 adipocytes.RBP was added to the medium of 3T3-L1 adipocytes or resistin-overexpressing adipocytes on day 0 of differentiation.Cell differentiation and lipid accumulation were determined by oil red O staining.The mRNA expressions of differentiation marker genes(pref-1,C/EBP?,FAS)and GLUT-4 gene were evaluated by RT-PCR.Triglyceride(TG)and free fatty acids(FFAs)in adipocytes were measured by colorimetric kit.Results(1)When 10-12mol/L RBP was applied,the percent of living cells was high and the shape was unchanged.(2)RBP had no effect on the differentiation of normal adipocytes,but significantly decreased the number of lipid droplets in resistin-overexpressing adipocytes without affecting the lipid droplets-presenting day.(3)C/EBP? and FAS expressions in resistin-overexpressing adipocytes were down-regulated after RBP was applied,without changing their expressions in normal adipocytes.(4)RBP had no effect on the cellular TG and FFAs levels in normal cells,whereas it can significantly decrease the levels in resistin-overexpressing adipocytes.(5)There was no difference in the expression of GLUT-4 gene between 3T3-L1 adipocytes and RBP-applied cells.Conclusions(1)RBP has no effect on the cell differentiation and lipid metabolism in normal 3T3-L1 adipocytes.(2)RBP can inhibit the cell differentiation and lipid metabolism of resistin-overexpressing 3T3-L1 cells.(3)RBP has no effect on the expression of GLUT-4 gene.