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Open Access 01-12-2016 | Research

Enhanced expression of the stemness-related factors OCT4, SOX15 and TWIST1 in ectopic endometrium of endometriosis patients

Authors: Katharina Proestling, Peter Birner, Sukirthini Balendran, Nadine Nirtl, Erika Marton, Gülen Yerlikaya, Lorenz Kuessel, Theresa Reischer, Rene Wenzl, Berthold Streubel, Heinrich Husslein

Published in: Reproductive Biology and Endocrinology | Issue 1/2016

Abstract

Background

Current evidence suggests that endometrial-derived stem cells, spilled in the peritoneal cavity via retrograde menstruation, are key players in the establishment of endometriotic lesions. The aim of this study was to determine the presence and distribution of the stemness-related factors OCT4, SOX15, TWIST1 and DCAMLK1 in women with and without endometriosis.

Methods

Immunohistochemical analysis was used to determine stromal and epithelial expression of OCT4, SOX15, TWIST1 and DCAMLK1 in endometriosis patient (EP) endometrium (n = 69) and endometriotic tissue (n = 90) and in control endometrium (n = 50). Quantitative Real-Time PCR of OCT4, SOX15 TWIST1 and DCAMLK1 was performed in paired samples of EP endometrium and endometriotic tissue. Co-immunofluorescence staining was performed for OCT4 and SOX15. For statistical analyses we used unpaired t-test, Fisher combination test and Spearman test. For paired analyses, paired t-test and McNemar test were used.

Results

We detected a significant correlation between the expression of the established stem cell marker OCT4 and the stemness-related markers SOX15 (p < 0.001) and TWIST1 (p = 0.002) but not DCAMLK1. We showed a colocalization of SOX15 and OCT4 in epithelial and stromal cells of endometriotic tissue by coimmunofluorescence. A concordant expression of OCT4 and SOX15 in the same sample was observed in epithelial cells of the endometriotic tissue (71.7%). The expression of stemness-related factors was not associated with proliferative or secretory phase of the menstrual cycle in endometriosis patients but was found to be differentially expressed during the menstrual cycle in the control group. Increased expression of epithelial OCT4, SOX15 and TWIST1 was detected in endometriotic tissue compared to EP endometrium in paired (p = 0.021, p < 0.001 and p < 0.001) and unpaired analysis (p = 0.040, p < 0.001 and p = 0.001).

Conclusion

Our findings support the hypothesis that upregulation of stem cell-related factors contribute to the establishment of endometriotic lesions.

Trial registration

The study was approved by the institutional review board (545/2010 on 6th of May 2014) of the Medical University of Vienna (http://ethikkommission.meduniwien.ac.at/fileadmin/ethik/media/dokumente/register/alle_2010.pdf).

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Giudice LC, Kao LC. Endometriosis. Lancet. 2004;364:1789–99.CrossRefPubMed

Sampson JA. Metastatic or embolic endometriosis, due to the menstrual dissemination of endometrial tissue into the venous circulation. Am J Pathol. 1927;3:93–110. 143.PubMedPubMedCentral

Maruyama T, Yoshimura Y. Molecular and cellular mechanisms for differentiation and regeneration of the uterine endometrium. Endocr J. 2008;55:795–810.CrossRefPubMed

Chan RW, Schwab KE, Gargett CE. Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod. 2004;70:1738–50.CrossRefPubMed

Gargett CE. Stem cells in gynaecology. Aust N Z J Obstet Gynaecol. 2004;44:380–6.CrossRefPubMed

Schwab KE, Chan RW, Gargett CE. Putative stem cell activity of human endometrial epithelial and stromal cells during the menstrual cycle. Fertil Steril. 2005;84 Suppl 2:1124–30.CrossRefPubMed

Figueira PG, Abrao MS, Krikun G, Taylor HS. Stem cells in endometrium and their role in the pathogenesis of endometriosis. Ann N Y Acad Sci. 2011;1221:10–7.CrossRefPubMedPubMedCentral

Sasson IE, Taylor HS. Stem cells and the pathogenesis of endometriosis. Ann N Y Acad Sci. 2008;1127:106–15.CrossRefPubMedPubMedCentral

Gargett BE, Chan RW. Endometrial stem/progenitor cells and proliferative disorders of the endometrium. Minerva Ginecol. 2006;58:511–26.PubMed

10.

Kim CM, Oh YJ, Cho SH, Chung DJ, Hwang JY, Park KH, Cho DJ, Choi YM, Lee BS. Increased telomerase activity and human telomerase reverse transcriptase mRNA expression in the endometrium of patients with endometriosis. Hum Reprod. 2007;22:843–9.CrossRefPubMed

11.

Starzinski-Powitz A, Zeitvogel A, Schreiner A, Baumann R. [Endometriosis--a stem cell disease?]. Zentralbl Gynakol. 2003;125:235–8.CrossRefPubMed

12.

Jimbo H, Hitomi Y, Yoshikawa H, Yano T, Momoeda M, Sakamoto A, Tsutsumi O, Taketani Y, Esumi H. Evidence for monoclonal expansion of epithelial cells in ovarian endometrial cysts. Am J Pathol. 1997;150:1173–8.PubMedPubMedCentral

13.

Meng X, Ichim TE, Zhong J, Rogers A, Yin Z, Jackson J, Wang H, Ge W, Bogin V, Chan KW, et al. Endometrial regenerative cells: a novel stem cell population. J Transl Med. 2007;5:57.CrossRefPubMedPubMedCentral

14.

Tamura M, Fukaya T, Murakami T, Uehara S, Yajima A. Analysis of clonality in human endometriotic cysts based on evaluation of X chromosome inactivation in archival formalin-fixed, paraffin-embedded tissue. Lab Investig. 1998;78:213–8.PubMed

15.

Tanaka M, Kyo S, Kanaya T, Yatabe N, Nakamura M, Maida Y, Okabe M, Inoue M. Evidence of the monoclonal composition of human endometrial epithelial glands and mosaic pattern of clonal distribution in luminal epithelium. Am J Pathol. 2003;163:295–301.CrossRefPubMedPubMedCentral

16.

Tanaka T, Nakajima S, Umesaki N. Cellular heterogeneity in long-term surviving cells isolated from eutopic endometrial, ovarian endometrioma and adenomyosis tissues. Oncol Rep. 2003;10:1155–60.PubMed

17.

Wu Y, Basir Z, Kajdacsy-Balla A, Strawn E, Macias V, Montgomery K, Guo SW. Resolution of clonal origins for endometriotic lesions using laser capture microdissection and the human androgen receptor (HUMARA) assay. Fertil Steril. 2003;79 Suppl 1:710–7.CrossRefPubMed

18.

Cho NH, Park YK, Kim YT, Yang H, Kim SK. Lifetime expression of stem cell markers in the uterine endometrium. Fertil Steril. 2004;81:403–7.CrossRefPubMed

19.

Matthai C, Horvat R, Noe M, Nagele F, Radjabi A, van Trotsenburg M, Huber J, Kolbus A. Oct-4 expression in human endometrium. Mol Hum Reprod. 2006;12:7–10.CrossRefPubMed

20.

Bentz EK, Kenning M, Schneeberger C, Kolbus A, Huber JC, Hefler LA, Tempfer CB. OCT-4 expression in follicular and luteal phase endometrium: a pilot study. Reproductive biology and endocrinology : RB&E. 2010;8:38.CrossRef

21.

Kato K, Yoshimoto M, Adachi S, Yamayoshi A, Arima T, Asanoma K, Kyo S, Nakahata T, Wake N. Characterization of side-population cells in human normal endometrium. Hum Reprod. 2007;22:1214–23.CrossRefPubMed

22.

Lynch L, Golden-Mason L, Eogan M, O'Herlihy C, O'Farrelly C. Cells with haematopoietic stem cell phenotype in adult human endometrium: relevance to infertility? Hum Reprod. 2007;22:919–26.CrossRefPubMed

23.

Tai MH, Chang CC, Kiupel M, Webster JD, Olson LK, Trosko JE. Oct4 expression in adult human stem cells: evidence in support of the stem cell theory of carcinogenesis. Carcinogenesis. 2005;26:495–502.CrossRefPubMed

24.

Park JH, Daheron L, Kantarci S, Lee BS, Teixeira JM. Human endometrial cells express elevated levels of pluripotent factors and are more amenable to reprogramming into induced pluripotent stem cells. Endocrinology. 2011;152:1080–9.CrossRefPubMedPubMedCentral

25.

Pacchiarotti A, Caserta D, Sbracia M, Moscarini M. Expression of oct-4 and c-kit antigens in endometriosis. Fertil Steril. 2011;95:1171–3.CrossRefPubMed

26.

Forte A, Schettino MT, Finicelli M, Cipollaro M, Colacurci N, Cobellis L, Galderisi U. Expression pattern of stemness-related genes in human endometrial and endometriotic tissues. Mol Med. 2009;15:392–401.CrossRefPubMedPubMedCentral

27.

Zeineddine D, Hammoud AA, Mortada M, Boeuf H. The Oct4 protein: more than a magic stemness marker. American J Stem Cells. 2014;3:74–82.

28.

Ng CK, Li NX, Chee S, Prabhakar S, Kolatkar PR, Jauch R. Deciphering the Sox-Oct partner code by quantitative cooperativity measurements. Nucleic Acids Res. 2012;40:4933–41.CrossRefPubMedPubMedCentral

29.

Revised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertility and sterility 1997, 67:817-821.

30.

Proestling K, Hebar A, Pruckner N, Marton E, Vinatzer U, Schreiber M. The Pro allele of the p53 codon 72 polymorphism is associated with decreased intratumoral expression of BAX and p21, and increased breast cancer risk. PLoS One. 2012;7, e47325.CrossRefPubMedPubMedCentral

31.

Fisher RA. Statistical Methods for Research Workers. Oliver and Boyd. Edingburgh: Oliver and Boyd; 1925.

32.

Bender R, Lange S. Adjusting for multiple testing--when and how? J Clin Epidemiol. 2001;54:343–9.CrossRefPubMed

33.

Gotte M, Wolf M, Staebler A, Buchweitz O, Kiesel L, Schuring AN. Aberrant expression of the pluripotency marker SOX-2 in endometriosis. Fertil Steril. 2011;95:338–41.CrossRefPubMed

34.

Gotte M, Wolf M, Staebler A, Buchweitz O, Kelsch R, Schuring AN, Kiesel L. Increased expression of the adult stem cell marker Musashi-1 in endometriosis and endometrial carcinoma. J Pathol. 2008;215:317–29.CrossRefPubMed

35.

Hwang JH, Oh JJ, Wang T, Jin YC, Lee JS, Choi JR, Lee KS, Joo JK, Lee HG. Identification of biomarkers for endometriosis in eutopic endometrial cells from patients with endometriosis using a proteomics approach. Mol Med Rep. 2013;8:183–8.PubMed

36.

Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol. 1984;64:151–4.PubMed

37.

D'Hooghe TM. Clinical relevance of the baboon as a model for the study of endometriosis. Fertil Steril. 1997;68:613–25.CrossRefPubMed

38.

Du H, Taylor HS. Contribution of bone marrow-derived stem cells to endometrium and endometriosis. Stem Cells. 2007;25:2082–6.CrossRefPubMed

39.

Chang JH, Au HK, Lee WC, Chi CC, Ling TY, Wang LM, Kao SH, Huang YH, Tzeng CR. Expression of the pluripotent transcription factor OCT4 promotes cell migration in endometriosis. Fertil Steril. 2013;99:1332–9. e1335.CrossRefPubMed

40.

Proestling K, Birner P, Gamperl S, Nirtl N, Marton E, Yerlikaya G, Wenzl R, Streubel B, Husslein H. Enhanced epithelial to mesenchymal transition (EMT) and upregulated MYC in ectopic lesions contribute independently to endometriosis. Reprod Biol Endocrinol. 2015;13:75.CrossRefPubMedPubMedCentral

41.

Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139:871–90.CrossRefPubMed

42.

Sureban SM, May R, Lightfoot SA, Hoskins AB, Lerner M, Brackett DJ, Postier RG, Ramanujam R, Mohammed A, Rao CV, et al. DCAMKL-1 regulates epithelial-mesenchymal transition in human pancreatic cells through a miR-200a-dependent mechanism. Cancer Res. 2011;71:2328–38.CrossRefPubMedPubMedCentral

43.

Nakanishi Y, Seno H, Fukuoka A, Ueo T, Yamaga Y, Maruno T, Nakanishi N, Kanda K, Komekado H, Kawada M, et al. Dclk1 distinguishes between tumor and normal stem cells in the intestine. Nat Genet. 2013;45:98–103.CrossRefPubMed

Title: Enhanced expression of the stemness-related factors OCT4, SOX15 and TWIST1 in ectopic endometrium of endometriosis patients
Authors: Katharina Proestling
Peter Birner
Sukirthini Balendran
Nadine Nirtl
Erika Marton
Gülen Yerlikaya
Lorenz Kuessel
Theresa Reischer
Rene Wenzl
Berthold Streubel
Heinrich Husslein
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Reproductive Biology and Endocrinology / Issue 1/2016
Electronic ISSN: 1477-7827
DOI: https://doi.org/10.1186/s12958-016-0215-4

At a glance: The STEP trials

Springer Medicine

Enhanced expression of the stemness-related factors OCT4, SOX15 and TWIST1 in ectopic endometrium of endometriosis patients

Abstract

Background

Methods

Results

Conclusion

Trial registration

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Trial registration

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