GENE-TRANSFER TO CULTURED HUMAN ENDOMETRIAL STROMAL CELLS - A MODEL TO STUDY CYCLOOXYGENASE-2 GENE-REGULATION

Objective: To compare and optimize conditions for gene transfer to hum an endometrial stromal cells derived from primary culture and to deter mine the effect of interleukin-1 beta (IL-1 beta) and phorbol 12-myris tate 13-acetate (PMA), a protein kinase C activator, on the promoter a ctivity of the human cyclooxygenase-2 (COX-2) gene. Design: Prospectiv e controlled study. Setting: Academic research laboratory. Patient(s): Women undergoing benign gynecologic surgery for indications other tha n endometrial diseases. Intervention(s): Endometrial stromal cells wer e used for transient transfection study. Main Outcome Measure(s): Luci ferase activity in transfected endometrial stromal cells. Result(s): G ene transfer mediated by cationic lipid was more efficient and more co nsistent. Lipofectamine, a polycationic lipid, yielded the highest eff iciency. Phorbol 12-myristate 13-acetate (30 nM) and IL-1 beta (100 ng /mL) increased COX-2 promoter activity by 2.6-fold and 2.2-fold, respe ctively. Conclusion(s): Induction of COX-2 by IL-1 beta and PMA sugges ts that COX-2 and prostaglandin have important roles in the growth and differentiation of endometrial stromal cells. This model can be used to explore the roles of different promoter regulatory elements in COX- 2 gene activation. (Fertil Steril(R) 1998;70:734-9. (C) 1998 by Americ an Society for Reproductive Medicine.).