title: Association between progesterone and estradiol-17beta treatment and protein expression of pgr and PGRMC1 in porcine luminal epithelial cells: a real-time cell proliferation approach.
abstract: The correct functionality (sensitivity and receptivity) of endometrial tissue is regulated by paracrine and endocrine pathways that activate several mediators or metabolic pathways and gene cascades. This study aimed to investigate the influence of E2 and P4 on progesterone receptor (PGR) and progesterone receptor membrane component 1 (PGRMC1) protein expression in porcine luminal epithelial cells and their influence on the proliferation of these cells in real-time. Surface uterine luminal epithelial cells were removed using sterile surgical blades from uterine horns of ten crossbred anestrus gilts. Following treatment with collagenase I, cells were separated and transferred into 48-well E-Plates for use in a realtime cell analyzer (RTCA). The luminal epithelial cells were cultured in vitro (IVC) in standard DMEM cell culture medium and incubated with E2 (10 pg/ml, 40 pg/ml, 500 pg/ml) and P4 (10 ng/ml, 40 ng/ ml, 500 ng/ml). The cell proliferation index was analyzed after 0-240 h, 0-120 h, 120-240 h. After using the RTCA analysis we found increased proliferation of luminal epithelial cells after treatment of low doses of P4 (10 and 40 ng/ml), (P < 0.001). Higher doses of P4 led to decrease of proliferation (P < 0.001). Conversely, higher doses of E2 (500 pg/ml) increased the proliferation index as compared to low doses (10 pg/ml) and control (P < 0.001). Confocal microscopic observations revealed that higher concentrations of E2 upregulate the expression of both PGR and PGRMC1. Additionally, P4 used in lower concentrations stimulated the expression of these receptors, too. Our study presents a new influence of E2 and P4 on the expression of PGR and PGRMC1 and on the real-time proliferation of porcine luminal epithelial cells. The relationship between PGR or PGRMC1 expression and the proliferation of luminal epithelial cells may be influenced (up- or down regulated) by E2 or P4 in a steroid type- and dose-dependent manner.
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: B
authors: B
authors: Kempisty
authors: Institute of Veterinary Sciences, Poznan University of Life Science, Poznań, Poland.
authors: K
authors: K
authors: Wojtanowicz-Markiewicz
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: A
authors: A
authors: Ziółkowska
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: J
authors: J
authors: Budna
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: S
authors: S
authors: Ciesiółka
authors: Department of Toxicology, Poznan University of Medical Sciences, Poznań, Poland.
authors: H
authors: H
authors: Piotrowska
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: A
authors: A
authors: Bryja
authors: Institute of Veterinary Sciences, Poznan University of Life Science, Poznań, Poland.
authors: P
authors: P
authors: Antosik
authors: Institute of Veterinary Sciences, Poznan University of Life Science, Poznań, Poland.
authors: D
authors: D
authors: Bukowska
authors: Institute of Reproductive Biology, Department of Experimental Reproductive Biology, Leibniz Institute for Farm Animal Biology, Dummerstorf, Germany.
authors: K
authors: K
authors: Wollenhaupt
authors: Department of Anatomy, Poznan University of Medical Sciences, Poznań, Poland.
authors: M
authors: M
authors: Bruska
authors: Institute of Reproductive Biology, Department of Experimental Reproductive Biology, Leibniz Institute for Farm Animal Biology, Dummerstorf, Germany.
authors: K P
authors: KP
authors: Brüssow
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: M
authors: M
authors: Nowicki
authors: Department of Histology and Embryology, Poznan University of Medical Science, Poznań, Poland.
authors: M
authors: M
authors: Zabel
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D004958:Estradiol / Q000378:metabolism / Q000494:pharmacology*; D005260:Female; D011374:Progesterone / Q000378:metabolism / Q000494:pharmacology*; D011980:Receptors, Progesterone / Q000378:metabolism*; D034421:Sus scrofa; D014599:Uterus / Q000166:cytology*
chemical_list: D011980:Receptors, Progesterone; D011374:Progesterone; D004958:Estradiol