title: Expression of exogenous human telomerase in cultures of endometrial stromal cells does not alter their hormone responsiveness.
abstract: In the human endometrium, stromal cells mediate the proliferative response of epithelial cells to the steroid hormones estrogen and progesterone. These stromal-epithelial interactions are readily studied in vitro by coculture of both cell types. A major impediment to such studies is the rapid senescence of normal stromal cells. To circumvent this problem, we tested whether human endometrial stromal cells immortalized by expressing a transduced human telomerase reverse transcriptase (TERT) subunit retained the ability to mediate hormonal control of epithelial proliferation in the coculture assay. We found that the telomerized stromal cells were very similar to the parental strain from which they were derived according to criteria of proliferation, karyotype, cellular localization of cytoskeletal markers and nuclear staining, and basal gene expression based on microarray analysis. We also showed that expression of estrogen and progesterone receptors, as assessed by immunodetection, was similar in both telomerized and parental stromal cells. Importantly, the telomerized stromal cells were shown in coculture assay to be as effective as normal stromal cells in regulating the proliferation of endometrial epithelial cells in response to estrogen or progesterone. The availability of these long-lived stromal cells may advance studies addressing the mechanistic, regulatory, and cell structural basis of stromal-epithelial interactions and hormonal responses in normal, preneoplastic, and neoplastic human endometrial tissue.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 27599, USA.
authors.forename: Claire S
authors.initials: CS
authors.lastname: Barbier
authors.forename: Karen A
authors.initials: KA
authors.lastname: Becker
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Troester
authors.forename: David G
authors.initials: DG
authors.lastname: Kaufman
mesh_terms: D015153:Blotting, Western; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D004717:Endometrium / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000096:biosynthesis* / Q000235:genetics; D047629:Estrogen Receptor beta / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000502:physiology; D011374:Progesterone / Q000494:pharmacology; D011388:Prolactin / Q000235:genetics / Q000502:physiology; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011980:Receptors, Progesterone / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology; D019098:Telomerase / Q000096:biosynthesis* / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D004268:DNA-Binding Proteins; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D017395:Plasminogen Activator Inhibitor 1; D011980:Receptors, Progesterone; D011374:Progesterone; D004958:Estradiol; D012313:RNA; D011388:Prolactin; C509186:TERT protein, human; D019098:Telomerase