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 human endometrial stromal cells derived from primary culture and to determine the effect of interleukin-1beta (IL-1beta) and phorbol 12-myristate 13-acetate (PMA), a protein kinase C activator. on the promoter activity of the human cyclooxygenase-2 (COX-2) gene.


METHODS
Prospective controlled study.


METHODS
Academic research laboratory.


METHODS
Women undergoing benign gynecologic surgery for indications other than endometrial diseases.


METHODS
Endometrial stromal cells were used for transient transfection study.


METHODS
Luciferase activity in transfected endometrial stromal cells.


RESULTS
Gene transfer mediated by cationic lipid was more efficient and more consistent. Lipofectamine, a polycationic lipid, yielded the highest efficiency. Phorbol 12-myristate 13-acetate (30 nM) and IL-1beta (100 ng/mL) increased COX-2 promoter activity by 2.6-fold and 2.2-fold, respectively.


CONCLUSIONS
Induction of COX-2 by IL-1beta and PMA suggests 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.
Huang
J C
JC
Department of Obstetrics, Gynecology and Reproductive Sciences, University of Texas Medical School at Houston, 77030, USA. JHUANG@OBG.MED.UTH.TMC.EDU
Dawood
M Y
MY
D000704:Analysis of Variance; D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D004717:Endometrium / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D018014:Gene Transfer Techniques; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D008565:Membrane Proteins; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D017154:Stromal Cells / Q000502:physiology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015533:Transcriptional Activation; D014162:Transfection
D007375:Interleukin-1; D007527:Isoenzymes; D008565:Membrane Proteins; D008156:Luciferases; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D013755:Tetradecanoylphorbol Acetate