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

Balamuthu Kadalmani, Hugh S. Taylor, Graciela Krikun, Kavitha Palanivel
In: Journal of Endometriosis and Pelvic Pain Disorders · 2013 · vol. 5(2) , pp. 77–85 · doi:10.5301/je.5000149 · W2085633238
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Endometriosis CD90 positive stromal progenitor cells showed increased expression of COX-2, StAR, CYP19A1, and altered apoptotic regulators compared to controls.

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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 +ve 90) 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.

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endometriosis

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