Abstract
Background
Inhibins and activins are important regulators of the female reproductive system. Recently, two novel inhibin subunits, named betaC and betaE, have been identified and shown to be expressed in several human tissues. However, only limited data on the expression of these novel inhibin subunits in normal human endometrial tissue and endometrial adenocarcinoma cell lines exist.
Materials and methods
Samples of proliferative and secretory human endometrium were obtained from five premenopausal, non-pregnant patients undergoing gynecological surgery for benign diseases. Normal endometrial tissue and Ishikawa endometrial adenocarcinoma cell lines were analyzed by immunohistochemistry, immunofluorescence and RT-PCR.
Results
Expression of the inhibin betaC and betaE subunits could be demonstrated at the protein level by means of immunohistochemical evaluation and at the transcriptional level by establishing a betaC- and betaE-specific RT-PCR analysis in normal human endometrial tissue and the parental Ishikawa cell line. Interestingly, in a highly de-differentiated subclone of the Ishikawa cell line lacking estrogen receptor expression, the expression of the inhibin-betaC subunit appeared strongly reduced.
Discussion
Here, we show for the first time that the novel inhibin/activin-betaC and -betaE subunits are expressed in normal human endometrium and the estrogen receptor positive human endometrial carcinoma cell line Ishikawa using RT-PCR and immunohistochemical detection methods. Interestingly, the Ishikawa minus cell line (lacking estrogen receptor expression) demonstrated no to minimal expression of the betaC subunit as observed with immunofluorescence and RT-PCR, suggesting a possible hormone- dependency of this subunit in human endometrial cancer cells. Moreover, because the Ishikawa cell line minus is thought to be a more malignant endometrial cell line than its estrogen receptor positive counterpart, inhibin-betaC subunit might be substantially involved in the pathogenesis and malignant transformation in human endometrium.
Similar content being viewed by others
References
Vale W, Rivier C, Hsueh A, Campen C, Meunier H, Bicsak T, Vaughan J, Corrigan A, Bardin W, Sawchenko P et al (1988) Chemical and biological characterization of the inhibin family of protein hormones. Recent Prog Horm Res 44:1–34
de Kretser DM, Hedger MP, Loveland KL, Phillips DJ (2002) Inhibins, activins and follistatin in reproduction. Hum Reprod Update 8:529–541
Xia Y, Schneyer AL (2009) The biology of activin: recent advances in structure, regulation and function. J Endocrinol 202:1–12
Hötten G, Neidhardt H, Schneider C, Pohl J (1995) Cloning of a new member of the TGF-beta family: a putative new activin beta C chain. Biochem Biophys Res Commun 206:608–613
Oda S, Nishimatsu S, Murakami K, Ueno N (1995) Molecular cloning and functional analysis of a new activin beta subunit: a dorsal mesoderm-inducing activity in Xenopus. Biochem Biophys Res Commun 210:581–588
Fang J, Yin W, Smiley E, Wang SQ, Bonadio J (1996) Molecular cloning of the mouse activin beta E subunit gene. Biochem Biophys Res Commun 228:669–674
Hashimoto O, Tsuchida K, Ushiro Y, Hosoi Y, Hoshi N, Sugino H, Hasegawa Y (2002) cDNA cloning and expression of human activin betaE subunit. Mol Cell Endocrinol 194:117–122
Fang J, Wang SQ, Smiley E, Bonadio J (1997) Genes coding for mouse activin beta C and beta E are closely linked and exhibit a liver-specific expression pattern in adult tissues. Biochem Biophys Res Commun 231:655–661
Vejda S, Cranfield M, Peter B, Mellor SL, Groome N, Schulte-Hermann R, Rossmanith W (2002) Expression and dimerization of the rat activin subunits betaC and betaE: evidence for the formation of novel activin dimers. J Mol Endocrinol 28:137–148
Mellor SL, Cranfield M, Ries R, Pedersen J, Cancilla B, de Kretser D, Groome NP, Mason AJ, Risbridger GP (2000) Localization of activin beta(A)-, beta(B)-, and beta(C)-subunits in humanprostate and evidence for formation of new activin heterodimers of beta(C)-subunit. J Clin Endocrinol Metab 85:4851–4858
Leung PH, Salamonsen LA, Findlay JK (1998) Immunolocalization of inhibin and activin subunits in human endometrium across the menstrual cycle. Hum Reprod 13:3469–3477
Petraglia F, Florio P, Luisi S, Gallo R, Gadducci A, Vigano P, Di Blasio AM, Genazzani AR, Vale W (1998) Expression and secretion of inhibin and activin in normal and neoplastic uterine tissues. High levels of serum activin A in women with endometrial and cervical carcinoma. J Clin Endocrinol Metab 83:1194–1200
Mylonas I, Jeschke U, Winkler L, Makovitzky J, Richter DU, Briese V, Friese K (2003) Immunohistochemical expression of inhibin-alpha in human endometrium and the in vitro secretion of inhibin, estradiol and cortisol in cultured human endometrial glandular cells. Arch Gynecol Obstet 268:142–150
Mylonas I, Makovitzky J, Fernow A, Richter DU, Jeschke U, Briese V, Gerber B, Friese K (2005) Expression of the inhibin/activin subunits alpha (alpha), beta-A (betaA) and beta-B (betaB) in benign human endometrial polyps and tamoxifen-associated polyps. Arch Gynecol Obstet 272:59–66
Mylonas I, Jeschke U, Wiest I, Hoeing A, Vogl J, Shabani N, Kuhn C, Schulze S, Kupka MS, Friese K (2004) Inhibin/activin subunits alpha, beta-A and beta-B are differentially expressed in normal human endometrium throughout the menstrual cycle. Histochem Cell Biol 122:461–471
Worbs S, Shabani N, Mayr D, Gingelmaier A, Makrigiannakis A, Kuhn C, Jeschke U, Kupka MS, Friese K, Mylonas I (2007) Expression of the inhibin/activin subunits (-alpha, -betaA and -betaB) in normal and carcinogenic endometrial tissue: Possible immunohistochemical differentiation markers. Oncol Rep 17:97–104
Jones RL, Findlay JK, Salamonsen LA (2006) The role of activins during decidualisation of human endometrium. Aust N Z J Obstet Gynaecol 46:245–249
Risbridger GP, Schmitt JF, Robertson DM (2001) Activins and inhibins in endocrine and other tumors. Endocr Rev 22:836–858
Mylonas I, Worbs S, Shabani N, Kuhn C, Kunze S, Schulze S, Dian D, Gingelmaier A, Schindlbeck C, Bruning A, Sommer H, Jeschke U, Friese K (2009) Inhibin-alpha subunit is an independent prognostic parameter in human endometrial carcinomas: Analysis of inhibin/activin-alpha, -betaA and -betaB subunits in 302 cases. Eur J Cancer 45:1304–1314
Mylonas I, Makovitzky J, Shabani N, Gingelmaier A, Dian D, Kuhn C, Schulze S, Kunze S, Jeschke U, Friese K (2009) Development and characterisation of an antibody for the immunohistochemical detection of inhibin/activin betaE (betaE) in normal human ovarian and placental tissue. Acta Histochem 111:366–371
Nishida M (2002) The Ishikawa cells from birth to the present. Hum Cell 15:104–117
Brüning A, Makovitzky J, Gingelmaier A, Friese K, Mylonas I (2009) The metastasis-associated genes MTA1 and MTA3 are abundantly expressed in human placenta and chorionic carcinoma cells. Histochem Cell Biol 132:33–38
Lau AL, Kumar TR, Nishimori K, Bonadio J, Matzuk MM (2000) Activin betaC and betaE genes are not essential for mouse liver growth, differentiation, and regeneration. Mol Cell Biol 20:6127–6137
Mylonas I, Makovitzky J, Hoeing A, Richter DU, Vogl J, Schulze S, Jeschke U, Briese V, Friese K (2006) Inhibin/activin subunits beta-A (-betaA) and beta-B (-betaB) are differentially localised in normal, hyperplastic and malignant human endometrial tissue. Acta Histochem 108:1–11
Beattie GM, Lopez AD, Bucay N, Hinton A, Firpo MT, King CC, Hayek A (2005) Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells 23:489–495
Welt CK (2002) The physiology and pathophysiology of inhibin, activin and follistatin in female reproduction. Curr Opin Obstet Gynecol 14:317–323
Jones RL, Salamonsen LA, Findlay JK (2002) Activin A promotes human endometrial stromal cell decidualization in vitro. J Clin Endocrinol Metab 87:4001–4004
Caniggia I, Lye SJ, Cross JC (1997) Activin is a local regulator of human cytotrophoblast cell differentiation. Endocrinology 138:3976–3986
Keelan JA, Zhou RL, Mitchell MD (2000) Activin A exerts both pro- and anti-inflammatory effects on human term gestational tissues. Placenta 21:38–43
Robson NC, Wei H, McAlpine T, Kirkpatrick N, Cebon J, Maraskovsky E (2009) Activin-A attenuates several human natural killer cell functions. Blood 113:3218–3225
Jones KL, de Kretser DM, Patella S, Phillips DJ (2004) Activin A and follistatin in systemic inflammation. Mol Cell Endocrinol 225:119–125
Phillips DJ, de Kretser DM, Hedger MP (2009) Activin and related proteins in inflammation: not just interested bystanders. Cytokine Growth Factor Rev 20:153–164
Kaitu’u-Lino TJ, Phillips DJ, Morison NB, Salamonsen LA (2009) A new role for activin in endometrial repair after menses. Endocrinology 150:1904–1911
Chen YG, Lui HM, Lin SL, Lee JM, Ying SY (2002) Regulation of cell proliferation, apoptosis, and carcinogenesis by activin. Exp Biol Med (Maywood) 227:75–87
Gold EJ, Zhang X, Wheatley AM, Mellor SL, Cranfield M, Risbridger GP, Groome NP, Fleming JS (2005) BetaA- and betaC-activin, follistatin, activin receptor mRNA and betaC-activin peptide expression during rat liver regeneration. J Mol Endocrinol 34:505–515
Chabicovsky M, Herkner K, Rossmanith W (2003) Overexpression of activin beta(C) or activin beta(E) in the mouse liver inhibits regenerative deoxyribonucleic acid synthesis of hepatic cells. Endocrinology 144:3497–3504
Vejda S, Erlach N, Peter B, Drucker C, Rossmanith W, Pohl J, Schulte-Hermann R, Grusch M (2003) Expression of activins C and E induces apoptosis in human and rat hepatoma cells. Carcinogenesis 24:1801–1809
Wada W, Medina JJ, Kuwano H, Kojima I (2005) Comparison of the function of the beta(C) and beta(E) subunits of activin in AML12 hepatocytes. Endocr J 52:169–175
Mellor SL, Ball EM, O’Connor AE, Ethier JF, Cranfield M, Schmitt JF, Phillips DJ, Groome NP, Risbridger GP (2003) Activin betaC-subunit heterodimers provide a new mechanism of regulating activin levels in the prostate. Endocrinology 144:4410–4419
Butler CM, Gold EJ, Risbridger GP (2005) Should activin betaC be more than a fading snapshot in the activin/TGFbeta family album? Cytokine Growth Factor Rev 16:377–385
Gold E, Jetly N, O’Bryan MK, Meachem S, Srinivasan D, Behuria S, Sanchez-Partida LG, Woodruff T, Hedwards S, Wang H, McDougall H, Casey V, Niranjan B, Patella S, Risbridger G (2009) Activin C antagonizes activin A in vitro and overexpression leads to pathologies in vivo. Am J Pathol 174:184–195
Takamura K, Tsuchida K, Miyake H, Tashiro S, Sugino H (2005) Activin and activin receptor expression changes in liver regeneration in rat. J Surg Res 126:3–11
Rodgarkia-Dara C, Vejda S, Erlach N, Losert A, Bursch W, Berger W, Schulte-Hermann R, Grusch M (2006) The activin axis in liver biology and disease. Mutat Res 613:123–137
Acknowledgments
We would like to thank Mrs. I. Krienke, Mrs. S. Kunze, Mrs S. Schulze, Mrs. S. Hoffmann and Mrs. I. Wiest for their excellent work with endometrial samples.
Conflict of interest statement
None.
Author information
Authors and Affiliations
Corresponding author
Additional information
T. Kimmich and A. Brüning contributed equally to this manuscript.
Rights and permissions
About this article
Cite this article
Kimmich, T., Brüning, A., Käufl, S.D. et al. Inhibin/activin-betaC and -betaE subunits in the Ishikawa human endometrial adenocarcinoma cell line. Arch Gynecol Obstet 282, 185–191 (2010). https://doi.org/10.1007/s00404-009-1310-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00404-009-1310-y