Abstract
The aspartyl-protease cathepsin D (cath-D) is overexpressed and hypersecreted by epithelial breast cancer cells and stimulates their proliferation. As tumor epithelial-fibroblast cell interactions are important events in cancer progression, we investigated whether cath-D overexpression affects also fibroblast behavior. We demonstrate a requirement of cath-D for fibroblast invasive growth using a three-dimensional (3D) coculture assay with cancer cells secreting or not pro-cath-D. Ectopic expression of cath-D in cath-D-deficient fibroblasts stimulates 3D outgrowth that is associated with a significant increase in fibroblast proliferation, survival, motility, and invasive capacity, accompanied by activation of the ras-MAPK pathway. Interestingly, all these stimulatory effects on fibroblasts are independent of cath-D proteolytic activity. Finally, we show that pro-cath-D secreted by cancer cells is captured by fibroblasts and partially mimics effects of transfected cath-D. We conclude that cath-D is crucial for fibroblast invasive outgrowth and could act as a key paracrine communicator between cancer and stromal cells, independently of its catalytic activity.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis / physiology
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Butadienes / pharmacology
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Cathepsin D / genetics
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Cathepsin D / metabolism
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Cathepsin D / physiology*
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Cell Enlargement / drug effects
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Cell Growth Processes / drug effects
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Cell Growth Processes / physiology
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Cell Line
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Cell Line, Tumor
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Cell Movement / drug effects
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Cell Movement / physiology*
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Cell Proliferation / drug effects
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Cell Survival / physiology
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Coculture Techniques
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Culture Media, Conditioned / pharmacology
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Endocytosis / drug effects
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Enzyme Inhibitors / pharmacology
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Enzyme Precursors / metabolism
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Enzyme Precursors / physiology
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Fibroblasts / cytology*
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Fibroblasts / drug effects
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Fibroblasts / ultrastructure
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Humans
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Mannosephosphates / pharmacology
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Mice
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Microscopy, Electron
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Mitogen-Activated Protein Kinase 1 / metabolism
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Mitogen-Activated Protein Kinase 3 / metabolism
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Neoplasm Invasiveness
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Neoplasms, Glandular and Epithelial / enzymology
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Neoplasms, Glandular and Epithelial / metabolism
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Neoplasms, Glandular and Epithelial / pathology
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Nitriles / pharmacology
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Paracrine Communication / physiology
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Phosphorylation / drug effects
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RNA, Small Interfering / genetics
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Transfection
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Wound Healing
Substances
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Butadienes
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Culture Media, Conditioned
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Enzyme Inhibitors
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Enzyme Precursors
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Mannosephosphates
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Nitriles
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RNA, Small Interfering
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U 0126
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mannose-6-phosphate
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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procathepsin D
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Cathepsin D