title: Effects of testosterone and 17β-oestradiol on expression of the G protein-coupled receptor P2Y12 in megakaryocytic DAMI cells.
abstract: P2Y12 is an important G protein-coupled receptor that is involved in ADP-induced platelet aggregation, which is essential for normal haemostasis. Gender differences in the incidence of cardiovascular disease have been proposed to be linked to the effects of sex hormones on cardiovascular-related genes. We examined the influences of testosterone and 17β-oestradiol on P2Y12 gene expression in megakaryocytic DAMI cell line. Altered levels of P2Y12 mRNA, protein and the cAMP-dependent vasodilator-stimulated phosphoprotein-Ser157 (VASP-Ser157) phosphorylation were investigated after treatment with 17β-oestradioal or testosterone as compared to the control groups. Quantitative real-time PCR revealed that the P2Y12 mRNA levels were increased by testosterone in a dose-dependent manner, whereas 17β-oestrodiol had no effect on P2Y12 gene expression. Induction of the P2Y12 protein by testosterone was found in Western blots of the proteins isolated from testosterone-treated cells. Testosterone-mediated P2Y12 expression was repressed at both the transcriptional and translational levels by the anti-androgen receptor bicalutamide. Treatment with testosterone also resulted in a decrease in the level of VASP-Ser157 phosphorylation, as compared to the control group. The decrease in the level of VASP-Ser157 phosphorylation was reversed by bicalutamide. These findings suggest a novel pathway for testosterone regulation of P2Y12 expression in a megakaryocytic DAMI cell line. Further studies using primary human megakaryocytes and platelets could be necessary to know the effect of hormones on the P2Y12 expression in circulating platelets.
authors: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Gaegum-Dong, Jin-Gu, Busan, Korea. 2sujun@inje.ac.kr
authors: Su-Jun
authors: SJ
authors: Lee
authors: Jeong-Ah
authors: JA
authors: Kwon
authors: Sun-Ah
authors: SA
authors: Cho
authors: Yazun B
authors: YB
authors: Jarrar
authors: Jae-Gook
authors: JG
authors: Shin
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000726:Androgen Antagonists / Q000494:pharmacology; D000813:Anilides / Q000494:pharmacology; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004958:Estradiol / Q000494:pharmacology*; D005786:Gene Expression Regulation; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D058912:Purinergic P2Y Receptor Agonists / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D060888:Real-Time Polymerase Chain Reaction; D058925:Receptors, Purinergic P2Y12 / Q000235:genetics* / Q000378:metabolism; D012737:Sex Factors; D015398:Signal Transduction; D013739:Testosterone / Q000494:pharmacology*; D014105:Tosyl Compounds / Q000494:pharmacology; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D000726:Androgen Antagonists; D000813:Anilides; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D009570:Nitriles; D010750:Phosphoproteins; D058912:Purinergic P2Y Receptor Agonists; D012333:RNA, Messenger; D058925:Receptors, Purinergic P2Y12; D014105:Tosyl Compounds; D000098662:Vasodilator-Stimulated Phosphoprotein; D013739:Testosterone; D004958:Estradiol; D000244:Adenosine Diphosphate; C053541:bicalutamide