miR-210 and GPD1L regulate EDN2 in primary and immortalized human granulosa-lutein cells.

Abstract

Endothelin-2 (EDN2), expressed at a narrow window during the periovulatory period, critically affects ovulation and corpus luteum (CL) formation. LH (acting mainly via cAMP) and hypoxia are implicated in CL formation; therefore, we aimed to elucidate how these signals regulate EDN2 using human primary (hGLCs) and immortalized (SVOG) granulosa-lutein cells. The hypoxiamiR, microRNA-210 (miR-210) was identified as a new essential player in EDN2 expression. Hypoxia (either mimetic compound-CoCl2, or low O2) elevated hypoxia-inducible factor 1A (HIF1A), miR-210 and EDN2 Hypoxia-induced miR-210 was suppressed in HIF1A-silenced SVOG cells, suggesting that miR-210 is HIF1A dependent. Elevated miR-210 levels in hypoxia or by miR-210 overexpression, increased EDN2 Conversely, miR-210 inhibition reduced EDN2 levels, even in the presence of CoCl2, indicating the importance of miR-210 in the hypoxic induction of EDN2 A molecule that destabilizes HIF1A protein, glycerol-3-phosphate dehydrogenase 1-like gene-GPD1L, was established as a miR-210 target in both cell types. It was decreased by miR-210-mimic and was increased by miR-inhibitor. Furthermore, reducing GPD1L by endogenously elevated miR-210 (in hypoxia), miR-210-mimic or by GPD1L siRNA resulted in elevated HIF1A protein and EDN2 levels, implying a vital role for GPD1L in the hypoxic induction of EDN2 Under normoxic conditions, forskolin (adenylyl cyclase activator) triggered changes typical of hypoxia. It elevated HIF1A, EDN2 and miR-210 while inhibiting GPD1L Furthermore, HIF1A silencing greatly reduced forskolin's ability to elevate EDN2 and miR-210. This study highlights the novel regulatory roles of miR-210 and its gene target, GPD1L, in hypoxia and cAMP-induced EDN2 by human granulosa-lutein cells.

• Authors: Ketan Shrestha, Adepeju Esther Onasanya, Iris Eisenberg, Noa Wigoda, Simcha Yagel, Ronit Yalu, Rina Meidan, Tal Imbar
• Journal: Reproduction (Cambridge, England) (Reproduction) Vol. 155 Issue 2 Pg. 197-205 (02 2018) ISSN: 1741-7899 [Electronic] England
• PMID: 29301980 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 Society for Reproduction and Fertility.

Chemical References

• Endothelin-2
• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• MIRN210 microRNA, human
• MicroRNAs
• RNA, Messenger
• GPD1L protein, human
• Glycerolphosphate Dehydrogenase

Topics

• Adult
• Cell Hypoxia
• Cells, Cultured
• Endothelin-2 (genetics, metabolism)
• Female
• Gene Expression Regulation
• Glycerolphosphate Dehydrogenase (genetics, metabolism)
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics, metabolism)
• Luteal Cells (cytology, metabolism)
• MicroRNAs (genetics)
• Ovulation
• RNA, Messenger (genetics, metabolism)