Human EG-VEGF ELISA Kit

Prokineticin-1 (PROK1) serves important roles in the pathogenesis of polycystic ovary syndrome (PCOS); however, the association between microRNA (miR)-28-5p and PROK1 remains unclear. In the present study, the roles of miR-28-5p and PROK1, and their interaction in PCOS were investigated. Rat ovary granule cells were transfected with miR-28-5p mimics, and PROK1 expression levels were measured by reverse transcription-quantitative PCR and western blotting. A dual-luciferase reporter assay was performed to determine the association between miR-28-5p and PROK1. Additionally, pcDNA-PROK1 was co-transfected into rat ovary granule cells with miR-28-5p mimics. Cell proliferation, apoptosis, cell cycle and the expression of signaling proteins were investigated using Cell Counting Kit-8 assays, 5-ethynyl-2'-deoxyuridine staining, flow cytometry and western blotting, respectively. PROK1 expression was suppressed in rat ovary granule cells by miR-28-5p mimics, but upregulated following transfection with miR-28-5p inhibitors. The dual-luciferase reporter assay revealed that miR-28-5p binds to the 3'-untranslated region of PROK1. Proliferation activity was increased in PROK1-overexpressing cells; this effect was eliminated by co-transfection with miR-28-5p mimics. PROK1-overexpressing rat ovary granule cells exhibited significantly suppressed cell apoptosis and a decreased number of cells in G1; miR-28-5p mimics reversed these effects. Western blotting revealed that the PI3K/AKT/mTOR signaling pathway was activated by PROK1. The present results suggested that miR-28-5p attenuated the progression of PCOS by targeting PROK1, which may promote the pathogenesis of PCOS via the PI3K/AKT/mTOR pathway, indicating that the miR-28-5p/PROK1 axis may be a potential therapeutic target for patients with PCOS.

Context: Prokineticin 1 (PROK1), also called endocrine gland-derived vascular endothelial growth factor, is a well-established regulator of endometrial receptivity and placental development. However, its clinical usefulness as a noninvasive predictive biomarker of embryo implantation is yet to be validated. Objective: The main objective of this article was to determine the relationship between PROK1 levels in the follicular fluid (FF) and fertilization culture media (FCM) and the reproductive outcome in patients who received a first conventional in vitro fertilization-embryo transfer. The secondary objective was to characterize the expression of PROK1 and its receptors (PROKRs) in the human follicular microenvironment. Design and Setting: We conducted a prospective study between January 2013 and June 2015 at the University Hospital of Grenoble. Patients: A total of 135 infertile in vitro fertilization patients and 10 women undergoing ovarian tissue cryopreservation were included. Interventions: The PROK1 concentration was measured by ELISA in FF and FCM collected on the day of oocyte retrieval and the day of the oocyte denudation step, respectively. Follicular expression of the PROK1/PROKR system was determined by immunohistochemistry, RT-quantitative PCR, and ELISA. Main Outcome Measure: Assessment of the clinical pregnancy rates was the main outcome. Results: FF and FCM PROK1 levels were significantly higher in the embryo implantation group (P < .001) and were predictive of subsequent embryo implantation (area under the receiver operating characteristic curve, 0.91 [95% confidence interval, 0.81-1.00], P = .001; and 0.88 [0.72-1.00], P = .001, respectively). FF and FCM PROK1 levels remain similar irrespective of the embryo morphokinetic parameters (P = .71 and P = .83, respectively). The PROK1/PROKR system is expressed during human folliculogenesis. Conclusions: PROK1 levels in FF and FCM could constitute new predictive noninvasive markers of successful embryo implantation in conventional in vitro fertilization-embryo transfer.