PD35-09 DIFFERENTIAL ACTIONS OF ERα AND ERβ VIA NON-GENOMIC SIGNALING IN HUMAN PROSTATE STEM-PROGENITOR CELLS

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2016PD35-09 DIFFERENTIAL ACTIONS OF ERα AND ERβ VIA NON-GENOMIC SIGNALING IN HUMAN PROSTATE STEM-PROGENITOR CELLS Shyama Majumdar, Neha Malhotra, Susan Kasper, Lishi Xie, Timothy Gauntner, Wen-yang Hu, and Gail Prins Shyama MajumdarShyama Majumdar More articles by this author , Neha MalhotraNeha Malhotra More articles by this author , Susan KasperSusan Kasper More articles by this author , Lishi XieLishi Xie More articles by this author , Timothy GauntnerTimothy Gauntner More articles by this author , Wen-yang HuWen-yang Hu More articles by this author , and Gail PrinsGail Prins More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1081AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Estrogens have been implicated in prostate development and cancer; however, specific roles of estrogen receptor-α and estrogen receptor-β are not well established. Using human prostate stem-progenitor cells we have previously shown that non-genomic pathways are involved in estrogen actions. Our present study sought to elucidate specific roles for ERα and ERβ via membrane initiated signaling in stem-progenitor cells. METHODS Human prostate stem-progenitor cells were enriched from primary prostate epithelial cell culture of young, disease-free donors using a 3D prostasphere (PS) model as previously described. Cells were labeled using ERα or ERβ antibodies and prostate stem cell markers CD49f and TROP2 followed by triple channel FACS to quantify ERα+/ERβ+ cell numbers. To explore ERα, the benign human prostate stem cell line WPE-stem, with extremely low levels of ERα and ERβ, was stably transfected with a lentiviral-ERα expression vector. The human prostate cancer stem-like cell line HuSLC (ERβ++, ERα-) was used to interrogate ERβ actions. Cells were exposed to 10 nM estradiol (E2) over a 15 to 60 minute time course +/- ICI 182,870 (ICI), an ERα/β antagonist. Western Blotting was used to quantify phosphorylation of target proteins. RESULTS FACS analysis of day 7 PS cells labeled for ERα or ERβ revealed 66% of day 7 PS cells as ERα+ and 40% as ERβ+. Among ERα or ERβ positive PS cells, 4% were Trop2+/CD49fhigh (stem-like cells) and 10-12% were Trop2+/CD49fmedium (early stage progenitor cells). PS exposed to 10 nM E2 showed sequential phosphorylation of Src, Erk1/2, p38, Akt and NFκB (p65) over 60 minutes. ICI attenuated Akt and Erk1/2 phosphorylation, confirming membrane bound ERs are involved in downstream signaling. E2 treatment of HuSLCs showed phosphorylation of Erk1/2 but not Akt, indicating that ERβ signals exclusively through the MAPK pathway in these cells. Conversely, E2 treatment of WPE-stem cells overexpressing ERα resulted in robust phosphorylation of Akt but lower levels of Erk1/2 phosphorylation suggesting that Akt activation may be more reliant on ERα signaling. CONCLUSIONS ERs trigger cellular responses via activation of signaling cascades that originate at the membrane (non-genomic signaling) in addition to nuclear signaling. The present findings reveal that human prostate stem-progenitor cells express both ERα and ERβ, which differentially activate membrane signaling cascades. This may lead to unique downstream actions that influence prostate stem-progenitor cell proliferation as well as lineage commitment decisions. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e847 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Shyama Majumdar More articles by this author Neha Malhotra More articles by this author Susan Kasper More articles by this author Lishi Xie More articles by this author Timothy Gauntner More articles by this author Wen-yang Hu More articles by this author Gail Prins More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...