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Phenotypic Modulation of Human Urinary Tract Stroma-derived Fibroblasts by Transforming Growth Factor β3

Lookup NU author(s): Professor Rakesh Heer, Dr Anastasia Hepburn, Dr Tim Cheek, Rob Pickard, Professor Hing Leung

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Abstract

OBJECTIVES Animal models have described critical roles for transforming growth factor beta (TGF beta) isoforms in modulating urinary tract stroma phenotype. TGF beta 3 is of particular interest because it may regulate TGF beta 1 and TGF beta 2 expression, but its modulatory affect has not been so well characterized in human cells. In this study, we aim to determine whether TGF beta 3 treatment induced differentiation of human urinary tract stroma-derived fibroblasts to a smooth muscle-like phenotype. METHODS We established cultures of human urinary tract stroma-derived fibroblasts and studied the effects of TGF beta 3 treatment using proliferation assays, cell cycle analysis, immunocytochemistry, and Western blotting for expression of differentiation marker and downstream regulators, and fura-2 fluorescence to study the effects on intracellular calcium. RESULTS TGF beta 3 treatment induced proliferation that peaked at 72 hours, followed by enhanced expression of alpha-smooth muscle actin (alpha-SMA) with a maximal 3.4-fold increase at 168 hours. TGF beta 3 treatment decreased resting [Ca2+](i) by 70% and caused a 95% decrease in stimulated internal Ca2+ release regulated by the sarcoplasmic/endoplasmic calcium-ATPase pump. These effects were associated with upregulation of nuclear activator of T cells -1 (NFAT), a known regulator of cell differentiation. CONCLUSIONS TGF beta 3 treatment causes a time-specific response in the presence of serum, whereby fibroblasts initially proliferate and subsequently differentiate to a smooth muscle-like phenotype. This sequence was associated with stabilization of [Ca2+](i) stores, suggesting a role in the induction of hyperplasia and reduction of contractility; phenomena associated with a number of urinary tract pathologies. UROLOGY 76: 509.e13-509.e20, 2010. (C) 2010 Elsevier Inc.

Publication metadata

Author(s): Heer R, Clarke N, Rigas AC, Cheek TR, Pickard R, Leung HY

Publication type: Article

Publication status: Published

Journal: Urology

Year: 2010

Volume: 76

Issue: 2

Pages: 509.e13-509.e20

Print publication date: 23/05/2010

ISSN (print): 0090-4295

ISSN (electronic): 1527-9995

Publisher: Elsevier

URL: http://dx.doi.org/10.1016/j.urology.2010.03.018

DOI: 10.1016/j.urology.2010.03.018

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