Objectives:Tibolone (Tib) exhibits progestagenic activities in addition to its tissue-specific estrogenic activities. The purpose of the current study was to determine the progestagenic actions of Tib and its metabolites using target genes known to be regulated by progestins in human endometrial glandular and stromal cells.
Methods:Human endometrial glandular and stromal cells were isolated from endometrial tissue fragments and separately incubated with Tib and its metabolites. Real-time polymerase chain reaction (PCR) was used to determine the mRNA content of 17βhydroxy steroid dehydrogenase (17βHSD, type 2) and sulfotransferase (SULT1E1) in endometrial glandular cells, and prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP1) in endometrial stromal cells.
Results:In glandular cells, Tib and Δ4-tibolone (Δ4Tib) significantly increased the content of 17βHSD and SULT1E1 mRNA. In stromal cells, Tib and Δ4Tib increased PRL mRNA (∼ 30% of the capacity compared to progesterone) and had little effect on IGFBP1 mRNA. Anti-progestin, RU486, reversed the induction of SULT1E1 and PRL by progesterone or Tib. Also, the two 3 hydroxyl tobolone metabolites, especially 3βOHTib, showed some progestagenic effects.
Conclusions:The data showed that Tib and Δ4Tib exhibited clear progestagenic effects in endometrial glandular cells by inducing 17βHSD and SULT1E1, while in stromal cells the response was weaker in the induction of PRL and had little effect on IGFBP1. In addition, the 3βOHTib metabolite expressed progestagenic activity. These disparate effects in two types of cells may be beneficial for maintaining endometrial cells in a quiescent state.
1. Kloosterboer HJ. Tibolone: A steroid with a tissue-specific mode of action. J Steroid Biochem Mol Biol2001;76:231-238.
2.
2. Modelska K, Cummings S. ibolone for postmenopausal women: Systematic review of randomized trials. J Clin Endocrinol Metab2002;87:16-23.
3.
3. Tang B, Markiewicz L, Kloosterboer HJ, Gurpide E. Human endometrial 3 beta-hydroxysteroid dehydrogenase/isomerase can locally reduce intrinsic estrogenic/progestagenic activity ratios of a steroidal drug (Org OD 14). J Steroid Biochem Mol Biol1993;45:345-351.
4.
4. de Gooyer M, Schoonen W, Verheul H, Kloosterboer H. Receptor profiling and endocrine interactions of tibolone. Steroids2003;68:21-31.
5.
5. Hanifi-Moghaddam P, Gielen SC, Kloosterboer HJ, et al. Molecular portrait of the progestagenic and estrogenic actions of tibolone: Behavior of cellular networks in response to tibolone. J Clin Endocrinol Metab2005;90:973-983.
6.
6. Markiewicz L, Gurpide E. In vitro evaluation of estrogenic, estrogen antagonistic and progestagenic effects of a steroidal drug (Org OD-14) and its metabolites on human endometrium. J Steroid Biochem1990;35:535-541.
7.
7. Schatz F, Kuczynski E, Kloosterboer HJ, et al. Tibolone and its metabolites enhance tissue factor and PAI-1 expression in human endometrial stromal cells: Evidence of progestogenic effects. Steroids2005;70:840-845.
8.
8. Tseng L, Gurpide E. Induction of human endometrial estradiol dehydrogenase by progestins. Endocrinology1975;97:825-833.
9.
9. Tseng L, Liu HC. Stimulation of arylsulfotransferase activity by progestins in human endometrium in vitro. J Clin Endocrinol Metab1981;53:418-421.
10.
10. Falany JL, Falany CN. Regulation of estrogen sulfotransferase in human endometrial adenocarcinoma cells by progesterone. Endocrinology1996;137:1395-1401.
11.
11. Zhang W, Mazella J, Kloosterboer HJ, Tseng L. Effects of tibolone on nuclear receptors in human endometrial cells. J Am Obstet Gynecol2006;195:97-102.
12.
12. Blok LJ, De Ruiter PE, Kuhne EC, et al. Progestogenic effects of tibolone on human endometrial cancer cells. J Clin Endocrinol Metab2003;88:2327-2334.
13.
13. Frost RA, Mazella J, Tseng L. Insulin-like growth factor binding protein-1 inhibits the mitogenic effect of insulin-like growth factors and progestins in human endometrial stromal cells. Biol Reprod1993;49:104-111.
14.
14. Lane B, Oxberry W, Mazella J, Tseng L. Decidualization of human endometrial stromal cells in vitro: Effects of progestin and relaxin on the ultrastructure and production of decidual secretory proteins. Hum Reprod1994;9:259-266.
15.
15. Tseng L, Mazella J. Endometrial cell specific gene activation during implantation and early pregnancy. Front Biosci2002;7:d1566-d1574.
16.
16. Bell SC, Jackson JA, Ashmore J, Zhu HH, Tseng L. Regulation of insulin-like growth factor-binding protein-1 synthesis and secretion by progestin and relaxin in long term cultures of human endometrial stromal cells. J Clin Endocrinol Metab1991;72:1014-1024.
17.
17. Zhu HH, Huang JR, Mazella J, Rosenberg M, Tseng L. Differential effects of progestin and relaxin on the synthesis and secretion of immunoreactive prolactin in long term culture of human endometrial stromal cells. J Clin Endocrinol Metab1990;71:889-899.
18.
18. Tseng L, Gao JG, Chen R, Zhu HH, Maszella J, Powell DR. Effect of progestin, antiprogestin, and relaxin on the accumulation of prolactin and insulin-like growth factor-binding protein-1 messenger ribonucleic acid in human endometrial stromal cells. Biol Reprod1992;47:441-450.
19.
19. Liu HC, Tseng L. Estradiol metabolism in isolated human endometrial epithelial glands and stromal cells. Endocrinology1979;104:1674-1681.
20.
20. Mazella J, Tang M, Tseng L. Disparate effects of relaxin and TGF{beta}1: Relaxin increases, but TGF{beta}1 inhibits, the relaxin receptor and the production of IGFBP-1 in human endometrial stromal/decidual cells. Hum Reprod2004;19:1513-1518.
21.
21. Schatz F, Lockwood CJ. Progestin regulation of plasminogen activator inhibitor type 1 in primary cultures of endometrial stromal and decidual cells. J Clin Endocrinol Metab1993;77:621-625.
22.
22. Chegini N, Tang XM, Dou Q. The expression, activity and regulation of granulocyte macrophage-colony stimulating factor in human endometrial epithelial and stromal cells. Mol Hum Reprod1999;5:459-466.
23.
23. Okada H, Sanezumi M, Nakajima T, Okada S, Yasuda K, Kanzaki H. Rapid down-regulation of CD63 transcription by progesterone in human endometrial stromal cells. Mol Hum Reprod1999;5:554-558.
24.
24. Liu HC, He ZY, Mele CA, Veeck LL, Davis O, Rosenwaks Z. Human endometrial stromal cells improve embryo quality by enhancing the expression of insulin-like growth factors and their receptors in cocultured human preimplantation embryos. Fertil Steril1999;71:361-367.
25.
25. Maruyama T, Yoshimura Y, Yodoi J, Sabe H. Activation of c-Src kinase is associated with in vitro decidualization of human endometrial stromal cells. Endocrinology1999;140:2632-2636.
26.
26. Gao J, Mazella J, Tang M, Tseng L. Ligand-activated progesterone receptor isoform hPR-A is a stronger transactivator than hPR-B for the expression of IGFBP-1 (insulin-like growth factor binding protein-1) in human endometrial stromal cells. Mol Endocrinol2000;14:1954-1961.
27.
27. Gao J, Mazella J, Suwanichkul A, Powell DR, Tseng L. Activation of the insulin-like growth factor binding protein-1 promoter by progesterone receptor in decidualized human endometrial stromal cells. Mol Cell Endocrinol1999;153:11-17.
28.
28. Tseng L, Mazella J, Goligorsky MS, Rialas CM, Stefano GB. Dopamine and morphine stimulate nitric oxide release in human endometrial glandular epithelial cells. J Soc Gynecol Investig2000;7:343-347.
29.
29. Tseng L, Zhu HH, Mazella J, Koistinen H, Seppala M. Relaxin stimulates glycodelin mRNA and protein concentrations in human endometrial glandular epithelial cells. Mol Hum Reprod1999;5:372-375.
30.
30. Tseng L, Gurpide E. Estradiol and 20alpha-dihydroprogesterone dehydrogenase activities in human endometrium during the menstrual cycle. Endocrinology1974;94:419-423.
31.
31. Tseng L, Stolee A, Gurpide E. Quantitative studies on the uptake and metabolism of estrogens and progesterone by human endometrium. Endocrinology1972;90:390-404.
32.
32. Vos RM, Krebbers SF, Verhoeven CH, Delbressine LP. The in vivo human metabolism of tibolone. Drug Metab Dispos2002;30:106-112.
33.
33. Falany JL, Macrina N, Falany CN. Sulfation of tibolone and tibolone metabolites by expressed human cytosolic sulfotransferases. J Steroid Biochem Mol Biol2004;88:383-391.
34.
34. Christian M, Pohnke Y, Kempf R, Gellersen B, Brosens JJ. Functional association of PR and CCAAT/enhancer-binding protein beta isoforms:Promoter-dependent cooperation between PR-B and liver-enriched inhibitory protein, or liver-enriched activatory protein and PR-A in human endometrial stromal cells. Mol Endocrinol2002;16:141-154.
35.
35. Gao J, Mazella J, Tseng L. Activation of human insulin-like growth factor binding protein-1 gene promoter by a distal regulatory sequence in a human endometrial adenocarcinoma cell line. Mol Endocrinol1995;9:1405-1412.
36.
36. Tang M, Mazella J, Zhu HH, Tseng L. Ligand activated relaxin receptor increases the transcription of IGFBP-1 and prolactin in human decidual and endometrial stromal cells. Mol Hum Reprod2005;11:237-243.
