title: Phospholipase D1 as a key enzyme for decidualization in human endometrial stromal cells.
abstract: Using primary cell cultures of human endometrial stromal cells (ES cells), we investigated the role of phospholipase D (PLD) in 8-Br-cAMP-induced decidualization, which involves morphological and biological differentiation processes. When treated with 0.5 mM 8-Br-cAMP for 12 days, ES cells were transformed into a decidualized morphology and produced significant amounts of prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Simultaneously, the activity and expression levels of PLD1 increased. In addition, removal of 8-Br-cAMP from decidualized ES cells restored the undifferentiated state, and this was accompanied by decreases in PLD1 promoter activity and PLD1 expression. Overexpression of dominant negative (DN)-PLD1 inhibited the morphological changes induced by 0.5 mM 8-Br-cAMP, whereas PLD1 overexpression induced morphological changes in the absence of 0.5 mM 8-Br-cAMP treatment. Moreover, knockdown of PLD1 by siRNA and blockage of PLD by treatment with 0.3% 1-butanol decreased PRL/IGFBP1 mRNA expression, whereas PLD1 overexpression increased PRL/IGFBP1 mRNA expression. Treatment of ES cells with phosphatidic acid (PA) for 3 days induced PRL mRNA expression and morphological changes, which implies that PA is an end-product of PLD activation-induced decidualization. In addition, pretreatment of ES cells with mepacrine decreased PRL/IGFBP1 expression and inhibited morphological change, whereas pretreatment with propranolol caused no changes, as compared to cAMP-treated cells, which suggests that PA induces decidualization through phospholipase A2 (PLA2G1B). Taken together, these results suggest that PLD1 regulates 8-Br-cAMP-induced decidualization through PLA2G1B, and that PLD1 upregulation is essential for the decidualization of ES cells.
authors.affiliation: Department of Biochemistry, College of Medicine, Hanyang University, Seoul 133-791, Korea.
authors.forename: Mee-Sup
authors.initials: MS
authors.lastname: Yoon
authors.forename: Jun Bon
authors.initials: JB
authors.lastname: Koo
authors.forename: Yong Geon
authors.initials: YG
authors.lastname: Jeong
authors.forename: Yong Seok
authors.initials: YS
authors.lastname: Kim
authors.forename: Jung Han
authors.initials: JH
authors.lastname: Lee
authors.forename: Hyae Jin
authors.initials: HJ
authors.lastname: Yun
authors.forename: Ki Sung
authors.initials: KS
authors.lastname: Lee
authors.forename: Joong-Soo
authors.initials: JS
authors.lastname: Han
mesh_terms: D020001:1-Butanol / Q000494:pharmacology; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D002478:Cells, Cultured; D003656:Decidua / Q000187:drug effects* / Q000502:physiology*; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005799:Genes, Dominant; D054500:Group IB Phospholipases A2 / Q000378:metabolism; D006801:Humans; D018970:Insulin-Like Growth Factor Binding Protein 1 / Q000096:biosynthesis / Q000235:genetics; D010712:Phosphatidic Acids / Q000494:pharmacology; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011388:Prolactin / Q000096:biosynthesis / Q000235:genetics; D011796:Quinacrine / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D017154:Stromal Cells / Q000187:drug effects / Q000502:physiology*; D015854:Up-Regulation
chemical_list: D004791:Enzyme Inhibitors; C032231:IGFBP1 protein, human; D018970:Insulin-Like Growth Factor Binding Protein 1; D010712:Phosphatidic Acids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D015124:8-Bromo Cyclic Adenosine Monophosphate; D020001:1-Butanol; D011388:Prolactin; D054500:Group IB Phospholipases A2; C516202:PLA2G1B protein, human; D010739:Phospholipase D; C106801:phospholipase D1; D011796:Quinacrine