Phosphatidylinositol 3-kinase function is required for transforming growth factor β-mediated epithelial to mesenchymal transition and cell migration

Andrei V. Bakin, Anne K. Tomlinson, Neil A. Bhowmick, Harold L. Moses, Carlos L. Arteaga

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Abstract

We have studied the role of phosphatidylinositol 3-OH kinase (PI3K)-Akt signaling in transforming growth factor β (TGFβ)-mediated epithelial to mesenchymal transition (EMT). In NMuMG mammary epithelial cells, exogenous TGFβ1 induced phosphorylation of Akt at Ser-473 and Akt in vitro kinase activity against GSK-3β within 30 rain. These responses were temporally correlated with delocalization of E-cadherin, ZO-1, and integrin β1 from cell junctions and the acquisition of spindle cell morphology. LY294002, an inhibitor of the p110 catalytic subunit of PI3K, and a dominant-negative mutant of Akt blocked the delocalization of ZO-1 induced by TGFβ1, whereas transfection of constitutively active p110 induced loss of ZO-1 from tight junctions. In addition, LY294002 blocked TGFβ-mediated C-terminal phosphorylation of Smad2. Consistent with these data, TGFβ-induced p3TP-Lux and p(CAGA)12-Lux reporter activities were inhibited by LY294002 and transiently expressed dominant-negative p85 and Akt mutants in NMuMG and 4T1 cells. Dominant-negative RhoA inhibited TGFβ-induced phosphorylation of Akt at Ser-473, whereas constitutively active RhoA increased the basal phosphorylation of Akt, suggesting that RhoA in involved in TGFβ-induced EMT. Finally, LY294002 and neutralizing TGFβ1 antibodies inhibited ligand-independent constitutively active Akt as well as basal and TGFβ-stimulated migration in 4T1 and EMT6 breast tumor cells. Taken together, these data suggest that PI3K-Akt signaling is required for TGFβ-induced transcriptional responses, EMT, and cell migration.

Original languageEnglish (US)
Pages (from-to)36803-36810
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number47
DOIs
StatePublished - Nov 24 2000

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Phosphatidylinositol 3-Kinase
Epithelial-Mesenchymal Transition
Transforming Growth Factors
Cell Movement
Phosphorylation
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Phosphatidylinositol 3-Kinases
Cadherins
Glycogen Synthase Kinase 3
Intercellular Junctions
Rain
Tight Junctions
Neutralizing Antibodies
Integrins
Transfection
Tumors
Catalytic Domain
Breast
Phosphotransferases
Epithelial Cells

ASJC Scopus subject areas

  • Biochemistry

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Phosphatidylinositol 3-kinase function is required for transforming growth factor β-mediated epithelial to mesenchymal transition and cell migration. / Bakin, Andrei V.; Tomlinson, Anne K.; Bhowmick, Neil A.; Moses, Harold L.; Arteaga, Carlos L.

In: Journal of Biological Chemistry, Vol. 275, No. 47, 24.11.2000, p. 36803-36810.

Research output: Contribution to journalArticle

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AU - Moses, Harold L.

AU - Arteaga, Carlos L.

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