AG 205, a progesterone receptor membrane component 1 antagonist, ablates progesterone's ability to block oxidative stress-induced apoptosis of human granulosa/luteal cells

Abstract The present studies were designed to determine whether progesterone (P4)-progesterone receptor membrane component 1 (PGRMC1) signaling is able to attenuate the apoptotic effects of oxidative stress induced by hydrogen peroxide (H2O2). To achieve this goal, freshly isolated human granulosa/ luteal cells were maintained in culture. After several passages, the cells were treated with H2O2, which induced apoptosis within 2.5 h, while simultaneous treatment with P4 attenuated the apoptotic action of H2O2. AG 205, a PGRMC1 antagonist, eliminated P4's ability to prevent H2O2-induced apoptosis. AG 205 neither affected PGRMC1's cytoplasmic localization nor its interaction with PGRMC2, but appeared to reduce its presence within the nucleus. AG 205 also (1) increased the monomeric and decreased the higher molecular weight forms of PGRMC1 (i.e., dimers/oligomers) and (2) altered the expression of several genes involved in apoptosis. The most dramatic change was an approximate 8-fold increase in Harakiri (Hrk) mRNA. However, AG 205 did not induce apoptosis in the absence of H2O2. Taken together, these observations suggest that the higher molecular weight forms of PGRMC1 likely account in part for PGRMC1's ability to suppress the expression of Hrk. Harakiri is a BH-3 only member of the B-cell lymphoma 2 (BCL2) family that promotes apoptosis by binding to and antagonizing the antiapoptotic action of BCL2- and BCL2-like proteins. It is likely then that PGRMC1's ability to suppress Hrk is part of the mechanism through which P4-PGRMC1 signaling preserves the viability of human granulosa/luteal cells. Summary Sentence The PGRMC1 antagonist, AG 205, blocks the ability of P4 to inhibit H2O2-induced apoptosis by disrupting PGRMC1 dimers/oligomers and inducing the expression of the apoptosis inducing BH3-only protein, Harakiri.

[1]  H. Handa,et al.  Cystathionine β-synthase and PGRMC1 as CO sensors. , 2016, Free radical biology & medicine.

[2]  F. Arnesano,et al.  The Effects of Chronic Lifelong Activation of the AHR Pathway by Industrial Chemical Pollutants on Female Human Reproduction , 2016, PloS one.

[3]  H. Handa,et al.  Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance , 2016, Nature Communications.

[4]  J. Peluso,et al.  Progestin and AdipoQ Receptor 7, Progesterone Membrane Receptor Component 1 (PGRMC1), and PGRMC2 and Their Role in Regulating Progesterone's Ability to Suppress Human Granulosa/Luteal Cells from Entering into the Cell Cycle1 , 2015, Biology of reproduction.

[5]  I. Lavrik,et al.  Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining. , 2015, Journal of immunological methods.

[6]  P. Thomas,et al.  Role of Pgrmc1 in estrogen maintenance of meiotic arrest in zebrafish oocytes through Gper/Egfr. , 2015, The Journal of endocrinology.

[7]  Erinna F. Lee,et al.  Bid chimeras indicate that most BH3-only proteins can directly activate Bak and Bax, and show no preference for Bak versus Bax , 2015, Cell Death and Disease.

[8]  H. Ahsan,et al.  The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update , 2015, Archives of Toxicology.

[9]  K. Hutt.  The role of BH3-only proteins in apoptosis within the ovary. , 2014, Reproduction.

[10]  J. Peluso,et al.  Progesterone Receptor Membrane Component-1 (PGRMC1) and PGRMC-2 Interact to Suppress Entry into the Cell Cycle in Spontaneously Immortalized Rat Granulosa Cells1 , 2014, Biology of reproduction.

[11]  K. Yoshinaga.  Progesterone and Its Downstream Molecules as Blastocyst Implantation Essential Factors , 2014, American journal of reproductive immunology.

[12]  J. Pru,et al.  Expression of Progesterone Receptor Membrane Component-2 Within the Immature Rat Ovary and Its Role in Regulating Mitosis and Apoptosis of Spontaneously Immortalized Granulosa Cells1 , 2014, Biology of reproduction.

[13]  S. Young,et al.  Homeostasis Imbalance in the Endometrium of Women with Implantation Defects: The Role of Estrogen and Progesterone , 2014, Seminars in Reproductive Medicine.

[14]  J. Pru,et al.  Non-canonical progesterone signaling in granulosa cell function. , 2014, Reproduction.

[15]  Peter Vandenabeele,et al.  Determination of apoptotic and necrotic cell death in vitro and in vivo. , 2013, Methods.

[16]  G. Häcker,et al.  BH3-only proteins are tail-anchored in the outer mitochondrial membrane and can initiate the activation of Bax , 2012, Cell Death and Differentiation.

[17]  J. Peluso,et al.  Evidence for a genomic mechanism of action for progesterone receptor membrane component-1 , 2012, Steroids.

[18]  J. Peluso,et al.  Progesterone regulation of progesterone receptor membrane component 1 (PGRMC1) sumoylation and transcriptional activity in spontaneously immortalized granulosa cells. , 2012, Endocrinology.

[19]  C. Simón,et al.  Efficiency and purity provided by the existing methods for the isolation of luteinized granulosa cells: a comparative study. , 2012, Human reproduction.

[20]  K. Roth,et al.  Deficiency of Pro-apoptotic Hrk Attenuates Programmed Cell Death in the Developing Murine Nervous System but Does Not Affect Bcl-x Deficiency-Induced Neuron Apoptosis , 2011, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[21]  F. Rizvi,et al.  Mitochondrial Dysfunction Links Ceramide Activated HRK Expression and Cell Death , 2011, PloS one.

[22]  X. Liu,et al.  Progesterone inhibits apoptosis in part by PGRMC1-regulated gene expression , 2010, Molecular and Cellular Endocrinology.

[23]  R. Craven,et al.  Progesterone Receptor Membrane Component 1 (Pgrmc1): A Heme-1 Domain Protein That Promotes Tumorigenesis and Is Inhibited by a Small Molecule , 2010, Journal of Pharmacology and Experimental Therapeutics.

[24]  T. Shioda,et al.  Progesterone receptor membrane component-1 regulates the development and Cisplatin sensitivity of human ovarian tumors in athymic nude mice. , 2009, Endocrinology.

[25]  M. Urbanek,et al.  The role of genetic variation in the lamin a/c gene in the etiology of polycystic ovary syndrome. , 2009, The Journal of clinical endocrinology and metabolism.

[26]  I. Ben‐Ami,et al.  EGF-like growth factors as LH mediators in the human corpus luteum. , 2008, Human reproduction.

[27]  S. Bione,et al.  Alterations in the expression, structure and function of progesterone receptor membrane component-1 (PGRMC1) in premature ovarian failure. , 2008, Human molecular genetics.

[28]  K. Phoenix,et al.  Regulation of ovarian cancer cell viability and sensitivity to cisplatin by progesterone receptor membrane component-1. , 2008, The Journal of clinical endocrinology and metabolism.

[29]  Jing Xu,et al.  Maternal exposure to polycyclic aromatic hydrocarbons diminishes murine ovarian reserve via induction of Harakiri. , 2007, The Journal of clinical investigation.

[30]  W. Sierralta,et al.  Features of natural and gonadotropin-releasing hormone antagonist-induced corpus luteum regression and effects of in vivo human chorionic gonadotropin. , 2007, Journal of Clinical Endocrinology and Metabolism.

[31]  G. Häcker,et al.  BH3-only proteins trigger cytochrome c release, but how? , 2007, Archives of biochemistry and biophysics.

[32]  M. Cahill.  Progesterone receptor membrane component 1: An integrative review , 2007, The Journal of Steroid Biochemistry and Molecular Biology.

[33]  Sten Orrenius,et al.  Mitochondria, oxidative stress and cell death , 2007, Apoptosis.

[34]  J. Peluso,et al.  Progesterone regulation of human granulosa/luteal cell viability by an RU486-independent mechanism. , 2006, The Journal of clinical endocrinology and metabolism.

[35]  G. Salvesen,et al.  Human inhibitor of apoptosis proteins: why XIAP is the black sheep of the family , 2006, EMBO reports.

[36]  Laurie Lamoreaux,et al.  Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry. , 2006, Journal of immunological methods.

[37]  J. Peluso,et al.  Progesterone membrane receptor component 1 expression in the immature rat ovary and its role in mediating progesterone's antiapoptotic action. , 2006, Endocrinology.

[38]  S. Willis,et al.  Life in the balance: how BH3-only proteins induce apoptosis. , 2005, Current opinion in cell biology.

[39]  E. Falkenstein,et al.  Classic and Non-classic Progesterone Receptors are both Expressed in Human Spermatozoa , 2005, Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme.

[40]  E. Falkenstein,et al.  Chemical modification and structural analysis of the progesterone membrane binding protein from porcine liver membranes , 2001, Molecular and Cellular Biochemistry.

[41]  R. Stouffer.  Progesterone as a mediator of gonadotrophin action in the corpus luteum: beyond steroidogenesis. , 2003, Human reproduction update.

[42]  B. Rueda,et al.  The corpus luteum: an ovarian structure with maternal instincts and suicidal tendencies. , 2002, Frontiers in bioscience : a journal and virtual library.

[43]  K. Sengoku,et al.  The soluble and membrane-anchored forms of heparin-binding epidermal growth factor-like growth factor appear to play opposing roles in the survival and apoptosis of human luteinized granulosa cells. , 2002, Molecular human reproduction.

[44]  J. Lydon,et al.  Reproductive functions of the progesterone receptor isoforms: lessons from knock-out mice , 2001, Molecular and Cellular Endocrinology.

[45]  B. O’Malley,et al.  Reproductive Functions of the Progesterone Receptor , 2000, The Journal of the Society for Gynecologic Investigation: JSGI.

[46]  J. Peluso,et al.  Progesterone mediates its anti-mitogenic and anti-apoptotic actions in rat granulosa cells through a progesterone-binding protein with gamma aminobutyric acidA receptor-like features. , 1998, Biology of reproduction.

[47]  N. Inohara,et al.  harakiri, a novel regulator of cell death, encodes a protein that activates apoptosis and interacts selectively with survival‐promoting proteins Bcl‐2 and Bcl‐XL , 1997, The EMBO journal.

[48]  B. O’Malley,et al.  Reproductive phenotypes of the progesterone receptor null mutant mouse , 1996, The Journal of Steroid Biochemistry and Molecular Biology.

[49]  S. Chun,et al.  Gonadal cell apoptosis. , 1996, Recent progress in hormone research.

[50]  M. Vega,et al.  Functional luteolysis in response to hydrogen peroxide in human luteal cells. , 1995, The Journal of endocrinology.

[51]  J. Peluso,et al.  Effect of in vivo gonadotropin treatment on the ability of progesterone, estrogen, and cyclic adenosine 5'-monophosphate to inhibit insulin-dependent granulosa cell mitosis in vitro. , 1995, Biology of reproduction.

[52]  J. Richards,et al.  Molecular aspects of hormone action in ovarian follicular development, ovulation, and luteinization. , 1988, Annual review of physiology.

[53]  C. Channing,et al.  Inhibin activity of preovulatory follicles of gonadotropin-treated and untreated women. , 1984, Fertility and sterility.

[54]  A. Decherney,et al.  The relationship between follicular fluid steroid concentration and successful fertilization of human oocytes in vitro. , 1984, Fertility and sterility.