Research questions: Does Progesterone primed ovarian stimulation (PPOS) protocol influence the microRNA expression profiles of granulosa cells of follicular fluid in infertile patients? Will adding progestins in the follicular phase affect follicular microenvironment?
Design: This was a prospective study with granulosa cells collected from patients who received PPOS protocol (n = 15) and who received gonadotropin releasing hormones (GnRH) antagonist protocol (n = 15). MicroRNA sequencing was set to find differently expressed microRNAs. The most significant statistically differential expressed miRNAs are chosen to be validated by quantitative real-time polymerase chain reaction (qRT-PCR) following by target genes prediction and enrichment analysis.
Results: The microRNA sequencing found 24 up-regulated miRNAs and 37 down-regulated miRNA among which 10 microRNAs (miR-4697-5p, miR-6788-5p, miR-6869-5p, miR-4644, miR-199a-5p, miR-6801-3p, miR-146b-5p, miR-4261, miR-199a-3p, miR-150-5p) can be detected in all sequencing samples and rank top enrichment scores in all microRNAs. The PCR validation showed that the down-regulation of miR-4261 and up-regulation of miR-6869-5p were consistent with the sequencing results. Putative genes were identified to be confidently predictive. The enrichment analysis showed that the most enriched gene ontology terms involves cell proliferation and apoptosis and the most enriched pathway included Hippo signaling pathway and Wnt signaling pathway.
Conclusions: PPOS protocol could up-regulate the expression of miR-6869-5p and down-regulate the expression of miR-4261 in granulosa cells of follicular fluid in infertile patients which involves a lot of cellular and molecular mechanisms requiring follow-up studies.
Keywords: Controlled ovarian stimulation; Progestin primed ovarian stimulation; microRNA expression.
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