Development and characterization of a simian virus 40 immortalized bovine endometrial stromal cell line.
In ruminants, interferon-tau (IFNtau) is the maternal recognition signal inhibiting prostaglandin (PG) F2alpha production by endometrial epithelial cells and stimulating interferon-stimulated genes in the stroma. Stromal cells mediate the action of progesterone on epithelial cells during pregnancy. Our working hypothesis is that IFNtau acts as a molecular switch that turns on PGE(2) production in endometrial stromal cells while suppressing PGF2alpha production from epithelial cells. In this report we document immortalization and functional characterization of a bovine stromal cell line from the caruncular region of the endometrium [caruncular stromal cell (CSC)]. Primary stromal cells were immortalized by nucleofection with simian virus 40 large T antigen and integrase. The resulting cell line, CSC, expresses stromal cell-specific vimentin, estrogen, and progesterone receptors, and is amenable for transient transfection. Basal and stimulated production of PGE2 is higher than PGF2alpha and associated with cyclooxygenase (COX) 2 expression. Phorbol myristate acetate (PMA) and IFNtau up-regulate COX2 and PG production in a dose-dependent manner. When added together, low concentrations of IFNtau inhibit PMA-induced COX2 expression; whereas this inhibition is lost at high concentrations. Expression of signal transducer and activator of transcription 1 is induced by IFNtau at all concentrations studied but is not modulated by PMA. Because expression of signal transducer and activator of transcription 1 does not exhibit the biphasic response to IFNtau, we investigated the p38 MAPK pathway using the selective inhibitor SB203580. Inhibition of the p38 MAPK pathway abolishes IFNtau action on PG production. In summary, CSC appears as a good stromal cell model for investigating the molecular mechanisms related to IFNtau action and PG production in the bovine.
Krishnaswamy
Narayanan
N
Centre Hospitalier Universitaire de Québec, Centre Hospitalier de l'Université Laval, Université Laval, Québec, Canada.
Chapdelaine
Pierre
P
Tremblay
Jacques P
JP
Fortier
Michel A
MA
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D015237:Dinoprost / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004717:Endometrium / Q000166:cytology / Q000502:physiology* / Q000821:virology*; D047628:Estrogen Receptor alpha / Q000235:genetics; D005260:Female; D007371:Interferon-gamma / Q000494:pharmacology; D011453:Prostaglandins / Q000096:biosynthesis; D011980:Receptors, Progesterone / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013539:Simian virus 40 / Q000502:physiology*; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology* / Q000821:virology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D047628:Estrogen Receptor alpha; D011453:Prostaglandins; D011980:Receptors, Progesterone; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D015237:Dinoprost; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate