Analysis of differential microRNA expression in patient with gallbladder stones through high-throughput sequencing technologies / 天津医药

ABSTRACT

Objective To detect the differential expression profile of microRNAs between patients with or without gall?bladder stone. Methods Samples from 30 patients with gallbladder stones (GS) and 30 without gallbladder stones (GP) were collected, in which microRNAs expression profiles were examined using high-throughput sequencing instrument Illumi?na HiSeq 2500. MicroRNA sequences were obtained and compared to Genebank and Rfam database for classification. Differ?entially expressed microRNAs were screened, and their target genes were predicted. Significant enrichment analysis of GO and KEGG were performed. Real-time quantitative PCR was performed on selected miRNAs in order to validate their expres?sion. Results Clean tags were obtained from both GS group (n=2 215 832) and GP group (n=1 424 770). A total of 17 mi?croRNAs were differentially expressed between GS and GP groups with statistical significance, among which 9 were up-regu?lated and 8 were down-regulated in GS group compared to those in GP group. GO (Gene ocology) analysis showed that target genes were enriched in ion binding and transport, apolipoprotein binding, calcium channel activity, protein kinase activity, steroid hormone biosynthesis and metabolism. KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis is shown for the target genes enriched in cancer related pathways, including WNT, HIPPO pathways. qRT-PCR validation of some differen?tially expressed miRNAs confirmed the result of high-throughput data analysis. Conclusion The differential expression levels of microRNAs may play an important role in occurrence and development of gallbladder stones.