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Inhibition of epithelial‐to‐mesenchymal transition (EMT) in MDCK cells by cyclic AMP
Author(s) -
Romano Silvia,
Murray Fiona,
Insel Paul A
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1087.5
Subject(s) - epithelial–mesenchymal transition , vimentin , activator (genetics) , protein kinase a , chemistry , cancer research , fibronectin , transforming growth factor , medicine , ibmx , microbiology and biotechnology , endocrinology , biology , kinase , cell , downregulation and upregulation , biochemistry , gene , immunohistochemistry , forskolin , stimulation
Tubulointerstitial fibrosis contributes to renal disease and ultimately renal failure. Epithelial‐to‐mesenchymal transition (EMT) can be induced by transforming growth factor beta (TGF‐β) and contribute to renal fibrosis. Previous data has shown that the second messenger cAMP can inhibit EMT in renal epithelial cells, however the mechanism for this inhibition has not been elucidated. We thus tested the hypothesis that cAMP acting via protein kinase A (PKA) and/or Exchange protein activated by cAMP (Epac) prevents EMT in Madin Darby canine kidney (MDCK) cells, a model of renal epithelia. We found that 72 hrs treatment with 8‐CPT‐cAMP (CPT, 50 μM), a stable cAMP analogue, and prostaglandin E2 (PGE2, 1 μM + 200 μM IBMX, a phosphodiesterase inhibitor), both of which activate PKA and Epac, reduced the expression of mesenchymal markers (e.g. α‐SMA, Fibronectin and Vimentin) in response to TGFβ (5ng/mL). N6‐Phenyl‐cAMP (N6), a selective PKA activator, but not 8‐pCPT‐2′‐O‐Me‐cAMP (8Me) an activator of Epac, was as effective as CPT in reducing mesenchymal markers. CPT, PGE2 and N6, but not 8Me, also decreased the expression of a number of transcription factors associated with EMT (e.g. ZEB1, ZEB2 and Slug). As the key mediator of cAMP‐promoted decrease in EMT in renal cells, PKA is thus a potential therapeutic target for pathological settings characterized by EMT. Supported by NIH