mRNA-Expression of ERα, ERβ, and PR in Clonal Stem Cell Cultures Obtained from Human Endometrial Biopsies
Corresponding Author
A. N. Schüring
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorJ. Braun
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorS. Wüllner
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorL. Kiesel
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorCorresponding Author
M. Götte
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorCorresponding Author
A. N. Schüring
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorJ. Braun
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorS. Wüllner
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorL. Kiesel
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorCorresponding Author
M. Götte
Department of Gynecology and Obstetrics University of Münster Medical Center 48149 Münster, Germany , uni-muenster.de
Search for more papers by this authorAbstract
Background. Proliferation and differentiation of the endometrium are regulated by estrogen and progesterone. The enormous regenerative capacity of the endometrium is thought to be based on the activity of adult stem cells. However, information on endocrine regulatory mechanisms in human endometrial stem cells is scarce. In the present study, we investigated the expression of ERα, ERβ, and PR in clonal cultures of human endometrial stem cells derived from transcervical biopsies. Methods. Endometrial tissue of 11 patients was obtained by transcervical biopsy. Stromal cell suspensions were plated at clonal density and incubated for 15 days. Expression of ERα, ERβ and PR was determined by qPCR prior to and after one cloning round, and normalized to 18 S rRNA expression. Results. Expression of ERα and ERβ was downregulated by 64% and 89%, respectively (P = 0.002 and P < 0.001). In contrast, PR was not significantly downregulated, due to a more heterogenous expression pattern. Conclusions. Culture of human endometrial stroma cells results in a downregulation of ERα and ERβ, while expression of PR remained unchanged in our patient collective. These results support the hypothesis that stem cells may not be subject to direct stimulation by sex steroids, but rather by paracrine mechanisms within the stem cell niche.
References
- 1 Gargett C. E., Schwab K. E., Zillwood R. M., Nguyen H. P. T., and Wu D., Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium, Biology of Reproduction. (2009) 80, no. 6, 1136–1145, 2-s2.0-66149112393, https://doi.org/10.1095/biolreprod.108.075226.
- 2 Tanaka M., Kyo S., Kanaya T., Yatabe N., Nakamura M., Maida Y., Okabe M., and Inoue M., Evidence of the monoclonal composition of human endometrial epithelial glands and mosaic pattern of clonal distribution in luminal epithelium, American Journal of Pathology. (2003) 163, no. 1, 295–301, 2-s2.0-0038306893.
- 3 Kim J. Y., Tavaré S., and Shibata D., Counting human somatic cell replications: methylation mirrors endometrial stem cell divisions, Proceedings of the National Academy of Sciences of the United States of America. (2005) 102, no. 49, 17739–17744, 2-s2.0-29144521716, https://doi.org/10.1073/pnas.0503976102.
- 4 Götte M., Wolf M., Staebler A., Buchweitz O., Kiesel L., and Schüring A. N., Aberrant expression of the pluripotency marker SOX-2 in endometriosis, Fertility and Sterility. (2011) 95, no. 1, 338–341, 2-s2.0-78650416731, https://doi.org/10.1016/j.fertnstert.2010.08.006.
- 5 Park J. H., Daheron L., Kantarci S., Lee B. S., and Teixeira J. M., Human endometrial cells express elevated levels of pluripotent factors and are more amenable to reprogramming into induced pluripotent stem cells, Endocrinology. (2011) 152, 1080–1089.
- 6 Schwab K. E., Hutchinson P., and Gargett C. E., Identification of surface markers for prospective isolation of human endometrial stromal colony-forming cells, Human Reproduction. (2008) 23, no. 4, 934–943, 2-s2.0-44249088606, https://doi.org/10.1093/humrep/den051.
- 7 Götte M., Wolf M., Staebler A., Buchweitz O., Kelsch R., Schüring A. N., and Kiesel L., Increased expression of the adult stem cell marker Musashi-1 in endometriosis and endometrial carcinoma, Journal of Pathology. (2008) 215, no. 3, 317–329, 2-s2.0-46849098625, https://doi.org/10.1002/path.2364.
- 8 Schüring A. N., Schulte N., Kelsch R., Röpke A., Kiesel L., and Götte M., Characterization of endometrial mesenchymal stem-like cells obtained by endometrial biopsy during routine diagnostics, Fertility and Sterility. (2011) 95, no. 1, 423–426, 2-s2.0-78650416095, https://doi.org/10.1016/j.fertnstert.2010.08.035.
- 9 Cervelló I., Gil-Sanchis C., Mas A. et al., Human endometrial side population cells exhibit genotypic, phenotypic and functional features of somatic stem cells, PLoS ONE. (2010) 5, no. 6, 2-s2.0-77955290480, https://doi.org/10.1371/journal.pone.0010964, e10964.
- 10 Taylor H. S., Endometrial cells derived from donor stem cells in bone marrow transplant recipients, Journal of the American Medical Association. (2004) 292, no. 1, 81–85, 2-s2.0-3042781155, https://doi.org/10.1001/jama.292.1.81.
- 11 Ikoma T., Kyo S., Maida Y., Ozaki S., Takakura M., Nakao S., and Inoue M., Bone marrow-derived cells from male donors can compose endometrial glands in female transplant recipients, American Journal of Obstetrics and Gynecology. (2009) 201, no. 6, e1–e8, 2-s2.0-70749135709, https://doi.org/10.1016/j.ajog.2009.07.026.
- 12 Gargett C. E. and Chan R. W. S., Endometrial stem/progenitor cells and proliferative disorders of the endometrium, Minerva Ginecologica. (2006) 58, no. 6, 511–526, 2-s2.0-33846540906.
- 13 Götte M., Greve B., Kelsch R. et al., The adult stem cell marker musashi-1 modulates endometrial carcinoma cell cycle progression and apoptosis via notch-1 and p21(WAF1/CIP1), International Journal of Cancer. (2011) 129, 2042–2049.
- 14 Pacchiarotti A., Caserta D., Sbracia M., and Moscarini M., Expression of oct-4 and c-kit antigens in endometriosis, Fertility and Sterility. (2011) 95, 1171–1173, 2-s2.0-78149311881, https://doi.org/10.1016/j.fertnstert.2010.10.029.
- 15 Kato K., Takao T., Kuboyama A., Tanaka Y., Ohgami T., Yamaguchi S., Adachi S., Yoneda T., Ueoka Y., Kato K., Hayashi S., Asanoma K., and Wake N., Endometrial cancer side-population cells show prominent migration and have a potential to differentiate into the mesenchymal cell lineage, American Journal of Pathology. (2010) 176, no. 1, 381–392, 2-s2.0-73949096245, https://doi.org/10.2353/ajpath.2010.090056.
- 16 Hubbard S. A., Friel A. M., Kumar B., Zhang L., Rueda B. R., and Gargett C. E., Evidence for cancer stem cells in human endometrial carcinoma, Cancer Research. (2009) 69, no. 21, 8241–8248, 2-s2.0-70350560603, https://doi.org/10.1158/0008-5472.CAN-08-4808.
- 17 Cervelló I. and Simón C., Somatic stem cells in the endometrium, Reproductive Sciences. (2009) 16, no. 2, 200–205, 2-s2.0-59849117813, https://doi.org/10.1177/1933719108329955.
- 18 Figueira P. G.M., Abrão M. S., Krikun G., and [email protected], Taylor H., Stem cells in endometrium and their role in the pathogenesis of endometriosis, Annals of the New York Academy of Sciences. (2011) 1221, no. 1, 10–17, https://doi.org/10.1111/j.1749-6632.2011.05969.x.
- 19 Chan R. W. S. and Gargett C. E., Identification of label-retaining cells in mouse endometrium, Stem Cells. (2006) 24, no. 6, 1529–1538, 2-s2.0-33747618211, https://doi.org/10.1634/stemcells.2005-0411.
- 20 Bulun S. E., Cheng Y. H., Pavone M. E., Xue Q., Attar E., Trukhacheva E., Tokunaga H., Utsunomiya H., Yin P., Luo X., Lin Z., Imir G., Thung S., Su E. J., and Kim J. J., Estrogen receptor-α, Estrogen receptor-β, and progesterone resistance in endometriosis, Seminars in Reproductive Medicine. (2010) 28, no. 1, 36–43, 2-s2.0-76249101923, https://doi.org/10.1055/s-0029-1242991.
- 21 Gründker C., Günthert A. R., and Emons G., Hormonal heterogeneity of endometrial cancer, Advances in Experimental Medicine and Biology. (2008) 630, 166–188, 2-s2.0-49649129978.
- 22 Götte M., Endometrial cells get side-tracked: side population cells promote epithelial-mesenchymal transition in endometrial carcinoma, American Journal of Pathology. (2010) 176, no. 1, 25–28, 2-s2.0-73949101948, https://doi.org/10.2353/ajpath.2010.090775.
- 23 Hyodo S., Matsubara K., Kameda K., and Matsubara Y., Endometrial injury increases side population cells in the uterine endometrium: a decisive role of estrogen, The Tohoku Journal of Experimental Medicine. (2011) 224, no. 1, 47–55, https://doi.org/10.1620/tjem.224.47.
- 24 Noyes R. W., Hertig A. T., and Rock J., Dating the endometrial biopsy, Fertility and Sterility. (1950) 1, 3–25.
- 25 Chan R. W. S., Schwab K. E., and Gargett C. E., Clonogenicity of human endometrial epithelial and stromal cells, Biology of Reproduction. (2004) 70, no. 6, 1738–1750, 2-s2.0-2642585666, https://doi.org/10.1095/biolreprod.103.024109.
- 26 Livak K. J. and Schmittgen T. D., Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method, Methods. (2001) 25, no. 4, 402–408, 2-s2.0-0035710746, https://doi.org/10.1006/meth.2001.1262.
- 27 Cervelló I., Mas A., Gil-Sanchis C., Peris L., Faus A., Saunders P. T.K., Critchley H. O.D., and Simón C., [email protected], Reconstruction of endometrium from human endometrial side population cell lines, PLoS ONE. (2011) 6, no. 6, https://doi.org/10.1371/journal.pone.0021221, e21221.
- 28 Gargett C. E., Uterine stem cells: what is the evidence?, Human Reproduction Update. (2007) 13, no. 1, 87–101, 2-s2.0-33845581196, https://doi.org/10.1093/humupd/dml045.
- 29 Wada-Hiraike O., Hiraike H., Okinaga H., Imamov O., Barros R. P. A., Morani A., Omoto Y., Warner M., and Gustafsson J. Å., Role of estrogen receptor α in uterine stroma and epithelium: insights from estrogen receptor α-/- mice, Proceedings of the National Academy of Sciences of the United States of America. (2006) 103, no. 48, 18350–18355, 2-s2.0-33845333006, https://doi.org/10.1073/pnas.0608861103.