Transcriptional cooperation between p53 and NF-κB p65 regulates microRNA-224 transcription in mouse ovarian granulosa cells

MicroRNAs (miRNAs) have been indicated to play key roles in ovarian follicular development. However, little is known about how the miRNA gene expression itself is regulated in the mammalian ovary. We previously reported that miR-224 is involved in TGF-beta 1-mediated follicular granulosa cell (GC) growth and estradiol (E-2) production by targeting Smad4. Here, the transcriptional regulation of miR-224 expression in GCs was further investigated. Our results showed that both the tumor suppressor gene p53 and NF-kappa B p65 subunit suppressed the TGF-beta 1-induced increase in pri-miR-224 expression in GCs. ChIP assays demonstrated that TGF-beta 1 enhanced the binding of p53 and p65 to the proximal promoter region of GABAA receptor a subunit (miR-224 host gene). p53 and p65 transcriptionally cooperated to inactivate the GABAA receptor epsilon subunit promoter. In addition, p53/p65 could up-regulate Smad4 expression by inhibiting its target miR-224 in GCs which contributed, at least partially, to the effects of miR-224 and Smad4 on GC proliferation and E-2 release. Our results provide new data about the interplay between transcription factors involved in GC proliferation and function by cooperatively regulating miRNA expression. (c) 2013 Elsevier Ireland Ltd. All rights reserved.