Inhibition of proliferation and differentiation by RU 486 in human endometrial stromal and epithelial cells in vitro

Summary:

The effects of progesterone and RU 486 on cellular proliferation and differentiation in long term cultures of mixed human endometrial cells were studied. The endometrial tissue was obtained from women with normal menstrual cycles who were undergoing hysterectomy for benign growths. Estradiol supplemented cultures were treated with progesterone and/or RU 486 for 27 days. Cell number was measured by crystal violet assay, and prolactin secretion was used as a marker of differentiation. Progesterone doubled the rate of proliferation, but the addition of RU 486 reduced it to baseline again. The gestagen increased prolactin secretion up to 30 times, while the addition of RU 486 suppressed it to baseline levels. When administered to cells that were pretreated with progesterone for 15 days RU 486 abolished the progesterone effects. RU 486 alone was without any effect. Our results indicate that (1) in vitro progesterone is essential for the initiation and maintenance of proliferation and differentiation of endometrial cells and (2) RU 486 acts as a pure progesterone antagonist in our culture model.

Human endometrial cell culture - proliferation - differentiation - RU 486

References

• 1 Berthois Y, Salat-Baroux J, Cornet D, De Brux J, Kopp F, Martin P M. A multiparametric analysis of endometrial estrogen and progesterone receptors after the postovulatory administration of mifepristone.  Fertil Steril. 55 547-554 1991; 
• 2 Bhargava-Periwal S, Farooq A, Bhargava V L, Bhatla N, Vij U, Murugesan K. Effect of antiprogestin RU 486 on the proliferation and metabolic activity of stromal cells separated from human endometrium.  Contraception. 53 127-131 1996; 
• 3 Burleigh D W, Williams R F, Gordon K, Hsiu J G, Hodgen G D. Screeing for antiproliferative actions of mifepristone. Differential endometrial responses of primates versus rats.  Contraception. 58 45-50 1998; 
• 4 Chen G, Huang J R, Mazella J, Tseng L. Long-term effects of progestin and RU 486 on prolactin production and synthesis in human endometrial stromal cells.  Hum Reprod. 4 355-358 1989; 
• 5 Daly D C, Maslar I A, Riddick D H. Prolactin production during in vitro decidualization of proliferative endometrium.  Am J Obstet Gynecol. 145 672-678 1983; 
• 6 Felix J C, Farahmand S. Endometrial glandular proliferation and estrogen receptor content during the normal menstrual cycle.  Contraception. 55 19-22 1997; 
• 7 Ferenczy A, Bertrand G, Gelfand M M. Proliferation kinetics of human endometrium during the normal menstrual cycle.  Am J Obstet Gynecol. 133 859-867 1997; 
• 8 Gemzell-Danielsson K, Westlund P, Johannisson E, Swahn M L, Bygdeman M, Seppälä M. Effect of low weekly doses of mifepristone on ovarian function and endometrial development.  Hum Reprod. 11 256-264 1996; 
• 9 Gentry M K, Dalrymple J M. Quantitative microtiter cytotoxicity assay for shigella toxin.  J Clin Microbiol. 12 361-366 1980; 
• 10 Gravanis A, Schaison G, George M, De Brux J, Satyaswaroop P G, Baulieu E E, Robel P. Endometrial and pituitary responses to the steroidal antiprogestin RU 486 in postmenopausal women.  J Clin Endocrinol Metab. 60 156-163 1985; 
• 11 Greb R R, Kiesel L, Selbmann A K, Wehrmann M, Hogen G D, Goodman Al, Wallwiener D. Disparate actions of mifepristone (RU 486) on glands and stroma in the primate endometrium.  Hum Reprod. 14 198-206 1999; 
• 12 Healy D L. Endometrial prolactin and implantation.  Baillieres Clin Obstet Gynecol. 5 95-105 1991; 
• 13 Heikinheimo O, Hsiu J G, Gordon K, Kim S, Williams R F, Gibbons W E, Hodgen G D. Endometrial effects of RU 486 in primates - antiproliferative action despite signs of estrogen action and increased Caclin-B expression.  J Steroid Biochem Mol Biol. 59 179-190 1996; 
• 14 Huang J R, Tseng L, Bischof P, Janne O A. Regulation of prolactin production by progestin, estrogen, and relaxin in human endometrial stromal cells.  Endocrinology. 121 2011-2017 1987; 
• 15 Irwin J C, Kirk D, King R J, Quigley M M, Gwatkin R B. Hormonal regulation of human endometrial stromal cells in culture: an in vitro model for decidualization.  Fertil Steril. 52 761-768 1989; 
• 16 Irwin J C, Utian W H, Eckert R L. Sex steroids and growth factors differentially regulate the growth and differentiation of cultured human endometrial stromal cells.  Endocrinology. 129 2385-2392 1991; 
• 17 Jabbour H N, Critchley H O, Boddy S C. Expression of functional prolactin receptors in nonpregnant human endometrium: janus kinase-2, signal transducer and activator of transcription-1 (STAT1), and STAT5 proteins are phosphorylated after stimulation with prolactin.  J Clin Endocrinol Metab. 83 2545-2553 1998; 
• 18 Jones R L, Critchley H O, Brooks J, Jabbour N H, McNeilly A S. Localization and temporal expression of prolactin receptor in human endometrium.  J Clin Endocrinol Metab. 83 258-262 1998; 
• 19 Kettel L M, Murphy A A, Morales A J, Ulmann A, Baulieu E E, Yen S SC. Treatment of endometriosis with the antiprogesterone mifepristone (RU 486).  Fertil Steril. 65 23-28 1996; 
• 20 Kiesel L, Mappes M. Regulation of endometrial differentiation by synthetic steroids in vitro.  Ann NY Acad Sci. 734 237-244 1994; 
• 21 Lobacco-Henri C, Descomps B, Thaler-Dao H. RU 38486 inhibits intracellular calcium mobilization and PGI2 release from human myometrium: mechanisms of action.  J Steroid Biochem Mol Biol. 59 63-73 1996; 
• 22 Mäentausta O, Svalander P, Gemzell-Danielsson K, Bygdeman M, Vihko R. The effects of an antiprogestin, mifepristone, and an antiestrogen, tamoxifen, on endometrial 17 beta-hydroxysteroid dehydrogenase and progestin and estrogen receptors during the luteal phase of the menstrual cycle: an immunohistochemical study.  J Clin Endocrinol Metab. 77 913-918 1993; 
• 23 Mappes M. Parakrine und autokrine Wirkmechanismen bei der Proliferation und Differenzierung von Endometrium- und Endometriosezellen in Kultur. Inauguraldissertation zur Erlangung des Doktorgrades scientiarum humanarum der Fakultät für klinische Medizin I der Medizinische Gesamtfakultät der Ruprecht-Karls-Universität Heidelberg 1993
• 24 Marions L, Danielsson K G, Swahn M L, Bygdeman M. Contraceptive efficacy of low doses of mifepristone.  Fertil Steril. 70 813-816 1998; 
• 25 Maslar I A, Riddick D H. Prolactin production by human endometrium during the normal menstrual cycle.  Am J Obstet Gynecol. 135 751-754 1979; 
• 26 Meyer M E, Pornon A, Ji J W, Bocquel M T, Chambon P, Gronemeyer H. Agonistic and antagonistic activities of RU486 on the functions of the human progesterone receptor.  EMBO J. 9 3923-3932 1990; 
• 27 Musgrove E A, Lee C S, Cornish A L, Swarbrick A, Sutherland R L. Antiprogestin inhibition of cell cycle progression in T-47D breast cancer cells is accompanied by induction of the cyclin-dependent kinase inhibitor p21.  Mol Endocrinol. 11 54-66 1997; 
• 28 Pecci A, Scholz A, Pelster D, Beato M. Progestins prevent apoptosis in a rat endometrial cell line and increases the ratio of bcl-XL to bcl-XS.  J Biol Chem. 272 11791-11798 1997; 
• 29 Perusquia M, Kubli-Garfias C. Progesterone-like relaxant effect of RU 486 in the rat myometrium.  Life Sci. 54 1501-1506 1994; 
• 30 Schneider C C, Gibb R K, Taylor D D, Wan T, Gercel-Taclor C. Inhibition of endometrial cancer cell lines by mifepristone (RU 486).  J Soc Gynecol Investig. 5 334-338 1998; 
• 31 Slayden O D, Brenner R M. RU 486 action after estrogen priming in the endometrium and oviducts of Rhesus monkeys (Macaca mullatta).  J Clin Endocrinol Metab. 78 440-448 1994; 
• 32 Swahn M L, Bygdeman M, Cekan S, Xing S, Masironi B, Johannisson E. The effect of RU 486 administered during the early luteal phase on bleeding pattern, hormonal parameters and endometrium.  Hum Reprod. 5 402-408 1990; 
• 33 Tseng L, Mazella J. Prolactin and its receptor in human endometrium.  Semin Reprod Endocrinol. 17 23-27 1999; 
• 34 Tung L, Mohamed M K, Hoeffler J P, Takimoto G S, Horwitz K B. Antagonist-occupied human progesterone B-receptors activate transcription without binding to progesterone response elements and are dominantly inhibited by A-receptors.  Mol Endocrinol. 7 1256-1265 1993; 
• 35 Viville B, Charnock-Jones D S, Sharkey A M, Wetzka B, Smith S K. Distribution of the A and B forms of the progesterone receptor messenger ribonucleic acid and protein in uterine leimyomata and adjacent myometrium.  Hum Reprod. 12 815-822 1997; 
• 36 Wolf J P, Hsiu J G, Anderson T L, Ulmann A, Baulieu E E, Hodgen G D. Noncompetitive antiestrogenic effect of RU 486 in blocking the estrogen stimulated luteinizing hormone surge and the proliferative action of estradiol on endometrium in castrated monkeys.  Fertil Steril. 52 1055-1060 1989; 
• 37 Xiao C W, Goff A K. Hormonal regulation of estrogen and progesterone receptors in cultured bovine endometrial cells.  J Reprod Fertil. 115 101-109 1999;