Research Article| Volume 171, ISSUE 2, P295-300, December 2013

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Inhibitory effects of androstenedione on endometrial cells: implications for poor reproductive outcome among women with androgen excess

  • Seung Bin Park
    Department of Obstetrics and Gynecology, Dong-A University, College of Medicine, Busan 602-715, South Korea
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  • Myoungseok Han
    Corresponding author at: Department of Obstetrics and Gynecology, Dong-A University, College of Medicine, Dongdaeshin-dong 3-ga, Seo-gu, Busan 602-715, South Korea.
    Department of Obstetrics and Gynecology, Dong-A University, College of Medicine, Busan 602-715, South Korea
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Published:October 17, 2013DOI:



      Androstenedione (A4) is an androgen that can be metabolized by aromatase to estrone, but the effects of A4 on endometrial cell proliferation either as an androgen or via conversion to estrogens are unknown. The aim of this study was to investigate A4 effects on Ishikawa cells in culture.

      Study design

      Ishikawa cells were treated with increasing concentrations of A4 (0–1000 pmol) for 4 days. Cell proliferation was measured by the (4,5-dimethylthiaxol-2-yi)-2,5-diphenyltetraxolium bromide (MTT) assay. Apoptosis was analyzed through Annexin-V/propidium iodide (PI) staining and flow cytometry: 17β-hydroxy steroid dehydrogenase type 1 (17β-HSD1) and aromatase mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR). Western blotting was used to detect cell signaling expressions of Akt/MAPK.


      A4 treatment (1 nM) decreased cell proliferation and increased apoptosis, as demonstrated by MTT and flow cytometry or related gene expression. The cellular responses induced by A4 treatment were mediated by activation of the Akt and MAPK signaling pathway. Treatment had no effect on 17β-HSD1 and aromatase expression.


      A4 treatment induced growth inhibition and apoptosis of Ishikawa cells through activation of the Akt/MAPK pathway. Effects of A4 on Ishikawa cells occurred in the absence of increased 17β-HSD1 and aromatase expression. These results imply that women with excessive androgen, such as polycystic ovary syndrome, experience poor reproductive outcomes through androgen-regulated mechanisms.


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        • Bahrke M.S.
        • Yesalis C.E.
        Abuse of anabolic androgenic steroids and related substances in sport and exercise.
        Curr Opin Pharmacol. 2004; 4: 614-620
        • Malaviya A.
        • Gomes J.
        Androstenedione production by biotransformation of phytosterols.
        Bioresour Technol. 2008; 99: 6725-6737
        • Kim S.K.
        • Han M.
        Know-how of the hormonal therapy and the effect of the male hormone on uterus in the female to male transsexuals.
        in: Ayman Al-Hendy M.S. Hysterectomy. Intech, 2012
        • Tuckerman E.M.
        • Okon M.A.
        • Li T.
        • Laird S.M.
        Do androgens have a direct effect on endometrial function? An in vitro study.
        Fertil Steril. 2000; 74: 771-779
        • Maliqueo M.A.
        • Quezada S.
        • Clementi M.
        • et al.
        Potential action of androstenedione on the proliferation and apoptosis of stromal endometrial cells.
        Reprod Biol Endocrinol. 2004; 2: 81
        • Smuc T.
        • Rizner T.L.
        Expression of 17beta-hydroxysteroid dehydrogenases and other estrogen-metabolizing enzymes in different cancer cell lines.
        Chem Biol Interact. 2009; 178: 228-233
        • Chang C.
        • Kokontis J.
        • Swift S.
        • Liao S.T.
        Molecular cloning and structural analysis of complementary DNA of human and rat androgen receptors.
        Prog Clin Biol Res. 1990; 322: 53-63
        • Chang C.S.
        • Kokontis J.
        • Liao S.T.
        Structural analysis of complementary DNA and amino acid sequences of human and rat androgen receptors.
        Proc Natl Acad Sci U SA. 1988; 85: 7211-7215
        • Lin Y.
        • Lu Z.
        • Kokontis J.
        • Xiang J.
        Androgen receptor primes prostate cancer cells to apoptosis through down-regulation of basal p21 expression.
        Biochem Biophys Res Commun. 2013; 430: 289-293
        • Sharma M.
        • Chuang W.W.
        • Sun Z.
        Phosphatidylinositol 3-kinase/Akt stimulates androgen pathway through GSK3beta inhibition and nuclear beta-catenin accumulation.
        J Biol Chem. 2002; 277: 30935-30941
        • Zhang G.
        • Gurtu V.
        • Kain S.R.
        • Yan G.
        Early detection of apoptosis using a fluorescent conjugate of annexin V.
        Biotechniques. 1997; 23: 525-531
      1. Toxicology and carcinogenesis studies of androstenedione (CAS No. 63-05-8) in F344/N rats and B6C3F1 mice (gavage studies).
        Natl Toxicol Program Tech Rep Ser. 2010; 1 (3-171 passim): 7-31
        • Brock B.J.
        • Waterman M.R.
        Biochemical differences between rat and human cytochrome P450c17 support the different steroidogenic needs of these two species.
        Biochemistry. 1999; 38: 1598-1606
        • Heinlein C.A.
        • Chang C.
        Androgen receptor in prostate cancer.
        Endocr Rev. 2004; 25: 276-308
        • Lin Y.
        • Kokontis J.
        • Tang F.
        • et al.
        Androgen and its receptor promote Bax-mediated apoptosis.
        Mol Cell Biol. 2006; 26: 1908-1916
        • Lovely L.P.
        • Appa Rao K.B.
        • Gui Y.
        • Lessey B.A.
        Characterization of androgen receptors in a well-differentiated endometrial adenocarcinoma cell line (Ishikawa).
        J Steroid Biochem Mol Biol. 2000; 74: 235-241
        • Blasberg M.E.
        • Robinson S.
        • Henderson L.P.
        • Clark A.S.
        Inhibition of estrogen-induced sexual receptivity by androgens: role of the androgen receptor.
        Horm Behav. 1998; 34: 283-293
        • Panet-Raymond V.
        • Gottlieb B.
        • Beitel L.K.
        • Pinsky L.
        • Trifiro M.A.
        Interactions between androgen and estrogen receptors and the effects on their transactivational properties.
        Mol Cell Endocrinol. 2000; 167: 139-150
        • Cardenas H.
        • Pope W.F.
        Attenuation of estrogenic effects by dihydrotestosterone in the pig uterus is associated with downregulation of the estrogen receptors.
        Biol Reprod. 2004; 70: 297-302
        • Manin M.
        • Baron S.
        • Goossens K.
        • et al.
        Androgen receptor expression is regulated by the phosphoinositide 3-kinase/Akt pathway in normal and tumoral epithelial cells.
        Biochem J. 2002; 366: 729-736
        • Altomare D.A.
        • Testa J.R.
        Perturbations of the AKT signaling pathway in human cancer.
        Oncogene. 2005; 24: 7455-7464
        • Kandel E.S.
        • Hay N.
        The regulation and activities of the multifunctional serine/threonine kinase Akt/PKB.
        Exp Cell Res. 1999; 253: 210-229
        • Kops G.J.
        • de Ruiter N.D.
        • De Vries-Smits A.M.
        • Powell D.R.
        • Bos J.L.
        • Burgering B.M.
        Direct control of the Forkhead transcription factor AFX by protein kinase B.
        Nature. 1999; 398: 630-634
        • Liu B.N.
        • Yan H.Q.
        • Wu X.
        • et al.
        Apoptosis induced by benzyl isothiocyanate in gefitinib-resistant lung cancer cells is associated with Akt/MAPK pathways and generation of reactive oxygen species.
        Cell Biochem Biophys. 2013; 66: 81-92
        • Jiang Y.
        • Chen C.
        • Li Z.
        • et al.
        Characterization of the structure and function of a new mitogen-activated protein kinase (p38beta).
        J Biol Chem. 1996; 271: 17920-17926
        • Crews C.M.
        • Erikson R.L.
        Extracellular signals and reversible protein phosphorylation: what to Mek of it all.
        Cell. 1993; 74: 215-217
        • Castro A.
        • Johnson M.C.
        • Anido M.
        • Cortinez A.
        • Gabler F.
        • Vega M.
        Role of nitric oxide and bcl-2 family genes in the regulation of human endometrial apoptosis.
        Fertil Steril. 2002; 78: 587-595
        • Croxtall J.D.
        • Elder M.G.
        • White J.O.
        Hormonal control of proliferation in the Ishikawa endometrial adenocarcinoma cell line.
        J Steroid Biochem. 1990; 35: 665-669
        • Navo M.A.
        • Smith J.A.
        • Gaikwad A.
        • Burke T.
        • Brown J.
        • Ramondetta L.M.
        In vitro evaluation of the growth inhibition and apoptosis effect of mifepristone (RU486) in human Ishikawa and HEC1A endometrial cancer cell lines.
        Cancer Chemother Pharmacol. 2008; 62: 483-489
        • Li A.
        • Felix J.C.
        • Minoo P.
        • Amezcua C.A.
        • Jain J.K.
        Effect of mifepristone on proliferation and apoptosis of Ishikawa endometrial adenocarcinoma cells.
        Fertil Steril. 2005; 84: 202-211