A molecular toolbox to study progesterone receptor signaling.
Hormone signaling
PGR
Progesterone
RANKL
Reporter constructs
WNT4
Journal
Journal of mammary gland biology and neoplasia
ISSN: 1573-7039
Titre abrégé: J Mammary Gland Biol Neoplasia
Pays: United States
ID NLM: 9601804
Informations de publication
Date de publication:
29 Nov 2023
29 Nov 2023
Historique:
received:
20
07
2023
accepted:
29
10
2023
medline:
30
11
2023
pubmed:
29
11
2023
entrez:
29
11
2023
Statut:
epublish
Résumé
Progesterone receptor (PR) signaling is required for mammary gland development and homeostasis. A major bottleneck in studying PR signaling is the lack of sensitive assays to measure and visualize PR pathway activity both quantitatively and spatially. Here, we develop new tools to study PR signaling in human breast epithelial cells. First, we generate optimized Progesterone Responsive Element (PRE)-luciferase constructs and demonstrate that these new reporters are a powerful tool to quantify PR signaling activity across a wide range of progesterone concentrations in two luminal breast cancer cell lines, MCF7 and T47D. We also describe a fluorescent lentiviral PRE-GFP reporter as a novel tool to visualize PR signaling at the single-cell level. Our reporter constructs are sensitive to physiological levels of progesterone. Second, we show that low background signaling, and high levels of PR expression are a prerequisite for robustly measuring PR signaling. Increasing PR expression by transient transfection, stable overexpression in MCF7 or clonal selection in T47D, drastically improves both the dynamic range of luciferase reporter assays, and the induction of endogenous PR target genes as measured by qRT-PCR. We find that the PR signaling response differs per cell line, target gene and hormone concentration used. Taken together, our tools allow a more rationally designed approach for measuring PR signaling in breast epithelial cells.
Identifiants
pubmed: 38019315
doi: 10.1007/s10911-023-09550-0
pii: 10.1007/s10911-023-09550-0
pmc: PMC10687192
doi:
Substances chimiques
Progesterone
4G7DS2Q64Y
Receptors, Progesterone
0
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
24Subventions
Organisme : Nederlandse Organisatie voor Wetenschappelijk Onderzoek
ID : OCENW.KLEIN.169
Informations de copyright
© 2023. The Author(s).
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