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PMID: 15843433 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Protein kinase A gating of a pseudopodial-located RhoA/ROCK/p38/NHE1 signal module regulates invasion in breast cancer cell lines.

Molecular biology of the cell ·Vol. 16 ·No. 7 ·2005-07-00 ·Pages 3117-27

Cardone RA, Bagorda A, Bellizzi A, Busco G, Guerra L, Paradiso A, Casavola V, Zaccolo M, Reshkin SJ

Abstract

Metastasis results from a sequence of selective events often involving interactions with elements of the tumor-specific physiological microenvironment. The low-serum component of this microenvironment confers increased motility and invasion in breast cancer cells by activating the Na+/H+ exchanger isoform 1 (NHE1). The present study was undertaken to characterize the signal transduction mechanisms underlying this serum deprivation-dependent activation of both the NHE1 and the concomitant invasive characteristics such as leading edge pseudopodia development and penetration of matrigel in breast cancer cell lines representing different stages of metastatic progression. Using pharmacological and genetic manipulation together with transport and kinase activity assays, we observe that the activation of the NHE1 and subsequent invasion by serum deprivation in metastatic human breast cells is coordinated by a sequential RhoA/p160ROCK/p38MAPK signaling pathway gated by direct protein kinase A phosphorylation and inhibition of RhoA. Fluorescence resonance energy transfer imaging of RhoA activity and immunofluorescence analysis of phospho-RhoA and NHE1 show that serum deprivation dynamically remodels the cell, forming long, leading edge pseudopodia and that this signal module is preferentially compartmentalized in these leading edge pseudopodia, suggesting a tight topographic relation of the signaling module to an invasion-specific cell structure.

MeSH Terms
Breast Neoplasms/pathology Cation Transport Proteins/metabolism Cell Line, Tumor Collagen/metabolism Culture Media, Serum-Free/pharmacology Cyclic AMP-Dependent Protein Kinases/metabolism Disease Progression Down-Regulation Drug Combinations Enzyme Activation Fluorescence Resonance Energy Transfer Gene Expression Regulation, Neoplastic Genetic Vectors Humans Hydrogen-Ion Concentration Intracellular Signaling Peptides and Proteins Laminin/metabolism Membrane Proteins/metabolism Microscopy, Fluorescence Models, Biological Neoplasm Invasiveness Neoplasm Metastasis Phosphorylation Protein Serine-Threonine Kinases/metabolism Proteoglycans/metabolism Pseudopodia/metabolism Serine/chemistry Signal Transduction Sodium-Hydrogen Exchanger 1 Sodium-Hydrogen Exchangers/metabolism Subcellular Fractions Time Factors Up-Regulation p38 Mitogen-Activated Protein Kinases/metabolism rho-Associated Kinases rhoA GTP-Binding Protein/metabolism
Chemicals
Cation Transport Proteins Culture Media, Serum-Free Drug Combinations Intracellular Signaling Peptides and Proteins Laminin Membrane Proteins Proteoglycans SLC9A1 protein, human Sodium-Hydrogen Exchanger 1 Sodium-Hydrogen Exchangers matrigel Serine Collagen Protein Serine-Threonine Kinases rho-Associated Kinases Cyclic AMP-Dependent Protein Kinases p38 Mitogen-Activated Protein Kinases rhoA GTP-Binding Protein
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Cardone Rosa A
Department of General and Environmental Physiology, University of Bari, 70126 Bari, Italy.
Bagorda Anna
Bellizzi Antonia
Busco Giovanni
Guerra Lorenzo
Paradiso Angelo
Casavola Valeria
Zaccolo Manuela
Reshkin Stephan J
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e04-10-0945
Published
2005-07-00
Epub
2005-00-20
Pages
3117-27
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1165397
Subset
IM
Grants
Telethon · TCP00089 · Italy
External Links
PubMed source DOI: 10.1091/mbc.e04-10-0945 PMC Full Text View journal details
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