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Abstract

Purpose

Endometriosis is an invasive gynecologic disease characterized by diminished apoptosis, sustained ectopic survival of dysfunctional endometrial cells and implantation of endometriotic lesions outside of the uterus. The purpose of this investigation was to determine the expression pattern of apoptotic and steroidogenic genes in the progenitor stem cell population of eutopic tissues from women with endometriosis and compare them to controls. The expression of these genes was determined in a subpopulation of endometrial cells that displayed CD90 positivity (CD^+ve90) and colony forming capacity.

Methods

We obtained endometrial samples from women with or without endometriosis. Endometrial stromal cells (ESCs) were isolated and cultured for 15 days. Purified ESCs were sorted by using a multipotent mesenchymal stromal cell multi-color flow cytometry kit. Single cell cloning was performed by serial dilution in 96-micro well plates. Fifteen days later, colonies were identified (CFUs). The colonies were chosen and cultured. mRNA expression of apoptotic genes, mitogen activated kinase 14 (MAPK14), nuclear factor kappa B (NFkB), steroidogenic acute regulatory protein (StAR), aromatase (CYP19A1) and cyclo-oxygenase-2 (COX-2) were determined by qRT-PCR. Protein levels of StAR, CYP19A1 and COX-2 were determined by western blotting.

Results

A subset of stromal cells derived from women with endometriosis were isolated and identified as progenitor stem cells based on their CD90 positivity and colony forming ability. The cells displayed increased levels of MAPK14, NFkB, COX-2, StAR and CYP19A1 both at the mRNA and protein level compared to stromal cells derived from controls. Similarly, pro-apoptotic molecules such as Bax were decreased whereas anti-apoptotic molecules such as Bcl2 were decreased at the mRNA level compared to stromal cells derived from controls.

Conclusions

CD90^+ve ESCs derived from women with endometriosis displayed markers suggesting stem cell-like properties and aberrant expression of apoptotic and steroidogenic enzymes which may contribute to sustained survival of these cells.

Keywords

Apoptosis COX-2 CYP19A1 Eutopic endometrium MAPK NFkB StAR Stromal cell progenitors

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References

Götte

, Wolf

, Staebler

Increased expression of the adult stem cell marker Musashi-1 in endometriosis and endometrial carcinoma. J Pathol 2008; 215 (3): 317–29.

Hopwood

, Levison

D.A.

Atrophy and apoptosis in the cyclical human endometrium. J Pathol 1976; 119 (3): 159–66.

Kokawa

, Shikone

, Nakano

Apoptosis in the human uterine endometrium during the menstrual cycle. J Clin Endocrinol Metab 1996; 81 (11): 4144–7.

Maruyama

, Masuda

, Ono

, Kajitani

, Yoshimura

Human uterine stem/progenitor cells: their possible role in uterine physiology and pathology. Reproduction 2010; 140 (1): 11–22.

Nagori

C.B.

, Panchal

S.Y.

, Patel

Endometrial regeneration using autologous adult stem cells followed by conception by in vitro fertilization in a patient of severe Asherman's syndrome. J Hum Reprod Sci 2011; 4 (1): 43–8.

Macer

M.L.

, Taylor

H.S.

Endometriosis and infertility: a review of the pathogenesis and treatment of endometriosis-associated infertility. Obstet Gynecol Clin North Am 2012; 39 (4): 535–49.

Cakmak

, Taylor

H.S.

Implantation failure: molecular mechanisms and clinical treatment. Hum Reprod Update 2012; 17 (2): 242–53.

Story

, Kennedy

Animal studies in endometriosis: a review. ILAR journal / National Research Council. ILAR J 2004; 45 (2): 132–8.

Nisolle

, Donnez

Peritoneal endometriosis, ovarian endometriosis, and adenomyotic nodules of the rectovaginal septum are three different entities. Fertil Steril 1997; 68 (4): 585–96.

10.

Bonte

, Chapron

, Vieira

Histologic appearance of endometriosis infiltrating uterosacral ligaments in women with painful symptoms. J Am Assoc Gynecol Laparosc 2002; 9 (4): 519–24.

11.

Itoga

, Matsumoto

, Takeuchi

Fibrosis and smooth muscle metaplasia in rectovaginal endometriosis. Pathol Int 2003; 53 (6): 371–5.

12.

Eskenazi

, Warner

M.L.

Epidemiology of endometriosis. Obstet Gynecol Clin North Am 1997; 24 (2): 235–58.

13.

Giudice

L.C.

, Kao

L.C.

Endometriosis. Lancet 2004; 364 (9447): 1789–99.

14.

Sharpe-Timms

K.L.

, Nabli

, Zimmer

R.L.

, Birt

J.A.

, Davis

J.W.

Inflammatory cytokines differentially up-regulate human endometrial haptoglobin production in women with endometriosis. Hum Reprod 2010; 25 (5): 1241–50.

15.

Taniguchi

, Kaponis

, Izawa

Apoptosis and endometriosis. Front Biosci (Elite Ed). 2011; 3: 648–62.

16.

Harada

, Kaponis

, Iwabe

Apoptosis in human endometrium and endometriosis. Hum Reprod Update 2004; 10 (1): 29–38.

17.

, Taylor

H.S.

Stem cells and female reproduction. Reprod Sci. 2009; 16 (2): 126–39.

18.

Sasson

I.E.

, Taylor

H.S.

Stem cells and the pathogenesis of endometriosis. Ann N Y Acad Sci. 2008; 1127 (1): 106–15.

19.

Shikone

, Kokawa

, Yamoto

, Nakano

Apoptosis of human ovary and uterine endometrium during the menstrual cycle. Horm Res 1997; 48 (Suppl 3): 27–34.

20.

Jabbour

H.N.

, Boddy

S.C.

Prostaglandin E2 induces proliferation of glandular epithelial cells of the human endometrium via extracellular regulated kinase 1/2-mediated pathway. J Clin Endocrinol Metab 2003; 88 (9): 4481–7.

21.

Jabbour

H.N.

, Kelly

R.W.

, Boddy

S.C.

Autocrine/paracrine regulation of apoptosis in epithelial cells by prostaglandin E2. Prostaglandins Leukot Essent Fatty Acids 2002; 67 (5): 357–63.

22.

Ohno

, Ohno

, Suzuki

, Soma

, Inoue

Cyclooxygenase-2 expression correlates with apoptosis and angiogenesis in endometrial cancer tissue. Anticancer Res 2007; 27(6A): 3765–70.

23.

Tsai

S.J.

, Wu

M.H.

, Lin

C.C.

, Sun

H.S.

, Chen

H.M.

Regulation of steroidogenic acute regulatory protein expression and progesterone production in endometriotic stromal cells. J Clin Endocrinol Metab 2001; 86 (12): 5765–73.

24.

Zeitoun

, Takayama

, Sasano

Deficient 17beta-hydroxysteroid dehydrogenase type 2 expression in endometriosis: failure to metabolize 17beta-estradiol. J Clin Endocrinol Metab 1998; 83 (12): 4474–80.

25.

Sun

H.S.

, Hsiao

K.Y.

, Hsu

C.C.

, Wu

M.H.

, Tsai

S.J.

Transactivation of steroidogenic acute regulatory protein in human endometriotic stromalcells is mediated by the prostaglandin EP2 receptor. Endocrinology 2003; 144 (9): 3934–42.

26.

M.H.

, Sun

H.S.

, Lin

C.C.

Distinct mechanisms regulate cyclooxygenase-1 and −2 in peritoneal macrophages of women with and without endometriosis. Mol Hum Reprod 2002; 8 (12): 1103–10.

27.

Wolff

E.F.

, Wolff

A.B.

, Hongling

, Taylor

H.S.

Demonstration of multipotent stem cells in the adult human endometrium by in vitro chondrogenesis. Reprod Sci. 2007; 14 (6): 524–33.

28.

Taylor

H.S.

The role of HOX genes in human implantation. Hum Reprod Update 2000; 6 (1): 75–9.

29.

Santamaria

, Massasa

E.E.

, Feng

, Wolff

, Taylor

H.S.

Derivation of insulin producing cells from human endometrial stromal stem cells and use in the treatment of murine diabetes. Mol Ther 2011; 19 (11): 2065–71.

30.

Taylor

H.S.

Endometrial cells derived from donor stem cells in bone marrow transplant recipients. JAMA 2004; 292 (1): 81–5.

31.

Sarno

J.L.

, Schatz

, Lockwood

C.J.

, Huang

S.T.

, Taylor

H.S.

Thrombin and interleukin-1beta regulate HOXA10 expression in human term decidual cells: implications for preterm labor. J Clin Endocrinol Metab 2006; 91 (6): 2366–72.

32.

, Taylor

H.S.

Contribution of bone marrow-derived stem cells to endometrium and endometriosis. Stem Cells 2007; 25 (8): 2082–6.

33.

Taylor

H.S.

Endometrial cells derived from donor stem cells in bone marrow transplant recipients. JAMA 2004; 292 (1): 81–5.

34.

Taylor

R.N.

, Yu

, Torres

P.B.

Mechanistic and therapeutic implications of angiogenesis in endometriosis. Reprod Sci. 2009; 16 (2): 140–6.

35.

Bulun

S.E.

, Yang

, Fang

Role of aromatase in endometrial disease. J Steroid Biochem Mol Biol 2001; 79 (1-5): 19–25.

36.

Fang

, Yang

, Gurates

Genetic or enzymatic disruption of aromatase inhibits the growth of ectopic uterine tissue. J Clin Endocrinol Metab 2002; 87 (7): 3460–6.

37.

Yang

, Fang

, Suzuki

Regulation of aromatase P450 expression in endometriotic and endometrial stromal cells by CCAAT/enhancer binding proteins (C/EBPs): decreased C/EBPbeta in endometriosis is associated with overexpression of aromatase. J Clin Endocrinol Metab 2002; 87 (5): 2336–45.

38.

Gurates

, Sebastian

, Yang

WT1 and DAX-1 inhibit aromatase P450 expression in human endometrial and endometriotic stromal cells. J Clin Endocrinol Metab 2002; 87 (9): 4369–77.

39.

Matsuzaki

, Murakami

, Uehara

, Canis

, Sasano

, Okamura

Expression of estrogen receptor alpha and beta in peritoneal and ovarian endometriosis. Fertil Steril 2001; 75 (6): 1198–205.

40.

Fechner

, Husen

, Thole

Expression and regulation of estrogen-converting enzymes in ectopic human endometrial tissue. Fertil Steril 2007; 88 (4)(Suppl): 1029–38.

41.

Hudelist

, Czerwenka

, Keckstein

Expression of aromatase and estrogen sulfotransferase in eutopic and ectopic endometrium: evidence for unbalanced estradiol production in endometriosis. Reprod Sci. 2007; 14 (8): 798–805.

42.

Bukulmez

, Hardy

D.B.

, Carr

B.R.

Androstenedione up-regulation of endometrial aromatase expression via local conversion to estrogen: potential relevance to the pathogenesis of endometriosis. J Clin Endocrinol Metab 2008; 93 (9): 3471–7.

43.

Ota

, Igarashi

, Sasaki

, Tanaka

Distribution of cyclooxygenase-2 in eutopic and ectopic endometrium in endometriosis and adenomyosis. Hum Reprod 2001; 16 (3): 561–6.

44.

M.H.

, Shoji

, Wu

M.C.

Suppression of matrix metalloproteinase-9 by prostaglandin E(2) in peritoneal macrophage is associated with severity of endometriosis. Am J Pathol 2005; 167 (4): 1061–9.

45.

Banu

S.K.

, Lee

, Speights

V.O.

Jr. , Starzinski-Powitz

, Arosh

J.A.

Cyclooxygenase-2 regulates survival, migration, and invasion of human endometriotic cells through multiple mechanisms. Endocrinology 2008; 149 (3): 1180–9.

46.

Meresman

G.F.

, Vighi

, Buquet

R.A.

, Contreras-Ortiz

, Tesone

, Rumi

L.S.

Apoptosis and expression of Bcl-2 and Bax in eutopic endometrium from women with endometriosis. Fertil Steril 2000; 74 (4): 760–6.

Atypical Expression of COX-2,StAR,CYP19A1 and Apoptotic Regulators in CD90 Positive Endometrial Stromal Cells from Women with Endometriosis

Abstract

Purpose

Methods

Results

Conclusions

Keywords

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References