Previous studies have reported that microRNA-764-3p (miR-764-3p) is one of the most up-regulated microRNAs (miRNAs) in TGF-β1-stimulated mouse ovarian granulosa cells. However, little is known about the roles and mechanisms of miR-764-3p in granulosa cell function during follicular development. In this study, we found that overexpression of miR-764-3p inhibited 17β-estradiol (E2) synthesis of granulosa cells through directly targeting steroidogenic factor-1 (SF-1). MiR-764-3p inhibited SF-1 by affecting its messenger RNA (mRNA) stability, which subsequently suppressed the expression levels of Cyp19a1 gene (aromatase, a downstream target of SF-1). In addition, SF-1 was involved in regulation of miR-764-3p-mediated Cyp19a1 expression in granulosa cells which contributed, at least partially, to the effects of miR-764-3p on granulosa cell E2 release. These results suggest that miR-764-3p functions to decrease steroidogenesis by targeting SF-1, at least in part, through inactivation of Cyp19a1. Taken together, our data provide mechanistic insights into the roles of miR-764-3p on E2 synthesis. Understanding of potential miRNAs affecting estrogen synthesis will help to diagnose and treat steroid-related diseases.
miR-764-3p 17β-estradiol Granulosa cell SF-1
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This work was supported by the following grants: the National Natural Science Foundation of China (81100399); Chongqing Yuzhong District Natural Science Foundation of China (20150117).
Compliance with ethical standards
The authors have nothing to disclose.
Ll.W. performed cell extraction, RNA extraction, gene expression, Western blotting, and drafted the manuscript. C.L. performed cell proliferation and estradiol analysis. R.L. performed cell culture. Yl.D. and Yx.T. performed plasmid construction and data analysis. C.T. revised the manuscript. Hb.Q. designed experiments and wrote this manuscript. All of the authors have read and approved the final version of the manuscript.
ESM 1Supplemental Figure S1. The efficacy of SF-1 siRNAs was evaluated by Western blotting after 48 h transfection with either si-NC or si-SF-1. Supplemental Table S1. List of primer pairs for real-time PCR. Supplemental Table S2. List of primer pairs for construction of vectors. (DOCX 37 kb)
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