Next Generation Sequencing Analysis of exosomes in follicle fluid of PCOS/non-PCOS patients
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ABSTRACT: Purpose: the goals of this study are to identify the transcriptome profiling of exosome derived from follicle fluid of PCOS patients by next-generation sequencing (NGS). Methods:the transcriptome profiling of exosome derived from follicular fluid of five PCOS and five control patients were generated by deep sequencing, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: the transcriptome profiling including circRNA, lncRNA and mRNA were comparatively analyzed in the two groups of exosomes (exo-PCOS and exo-control). We focus on the circRNAs profile for further analysis. A circRNA candidate was considered to be differentially expressed if its levels between both experiment groups showed a statistically significant difference (p<0.05) of at least two-fold. Based on this criterion, we identified 16 differentially expressed circRNAs (5 up-regulated and 11 down-regulated circRNAs) between exo-PCOS and exo-control, including 3 annotated exonic circRNAs and 13 novel ones. Clustering analysis based on the differentially expressed circRNAs clustered the exo-PCOS and the exo-control groups (control). Conclusion: the circRNA expression profile of exosome derived from follicular fluid of five PCOS and five control patients were determined by RNA sequencing technology. 16 circRNAs showed significantly different expression in the exosome of PCOS FF. GO and KEGG pathway analysis indicated that the parental genes of the candidate circRNAs were involved in ovarian steroidogenesis, aldosterone synthesis and secretion, thyroid hormone signaling pathway, ubiquitin mediated proteolysis, Jak-STAT signaling pathway, and endocytosis pathway.
ORGANISM(S): Homo sapiens
PROVIDER: GSE145461 | GEO | 2020/09/09
REPOSITORIES: GEO
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