N5 Endometrial Stromal Cell Line: A Model System to Study Decidual Prolactin Gene Expression

• Published in: In vitro cellular & developmental biology. Animal Vol. 35; no. 3; pp. 150 - 154
• Main Authors: Brar, A K, Kanda, Y, Kessler, C A, Cedars, M I, Handwerger, S
• Format: Journal Article
• Language: English
• Published: Germany Society for In Vitro Biology 01-03-1999
• Subjects: Bucladesine - pharmacology; Cell Line; Cell Line, Transformed; Cell lines; Cellular Models; Cultured cells; Decidua - cytology; Dinoprostone - pharmacology; Endometrium; Endometrium - cytology; Estradiol - pharmacology; Exons; Female; Gene Expression; Genetic vectors; Humans; Medroxyprogesterone Acetate - pharmacology; Messenger RNA; Models, Biological; Polymerase chain reaction; Prolactin - genetics; Promoter Regions, Genetic; Stromal cells; Stromal Cells - cytology; Transfection
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-999-0017-5

Prolactin gene expression in extrapituitary tissues, such as decidua and lymphocytes, is regulated by a distinct promoter approximately 6 kb upstream of the pituitary prolactin gene transcription start site. Here we describe studies in a human endometrial stromal cell line, N5, that was immortalized by transfection with an SV40 mutant and which expresses the prolactin gene driven by the extrapituitary promoter. The N5 cells have phenotypic features of primary cultures of decidualized human endometrial stromal cells and secrete low levels of prolactin and insulin-like growth factor binding protein-1 (IGFBP-1), both of which are markers of decidualized endometrial stromal cells. As in primary cultures of endometrial stromal cells, treatment of N5 cells with progesterone and estradiol alone or in combination with prostaglandin E2stimulated the synthesis and release of prolactin. Transient transfection of the N5 cells with an expression vector containing - 2927/+ 66 bp of the decidual prolactin promoter coupled to a luciferase reporter gene resulted in a 20 to 25-fold increase in luciferase activity, a magnitude similar to that which occurs in primary cultures of endometrial stromal cells decidualized in vitro by treatment with progesterone and estradiol. Luciferase expression levels were similar in untreated N5 cells and N5 cells treated with progesterone and estradiol. Taken together, these results indicate that the N5 human endometrial stromal cell line has phenotypic characteristics of normal decidualized stromal cells and is a useful model to study regulation of decidual prolactin gene expression.