- Poster Presentation
- Open Access
Lack of evidence for nuclear IGFBP5 in mammary epithelial cells
© BioMed Central 2005
- Published: 17 June 2005
- Signal Peptide
- Nuclear Localization
- Mammary Epithelial Cell
- Mammary Epithelial
IGFBP5 plays a role in mediating the effects of IGFs, which are important in mammary gland development  and carcinogenesis . IGF-independent effects of IGFBP5 have also been described and it has been postulated that these are at least partially mediated via IGFBP5 localized in the nucleus [3, 4].
The cellular localization of IGFBP5 was analyzed by confocal microscopy after either applying exogenous fluorescent-labeled recombinant protein or applying immunostaining of cells ectopically expressing IGFBP5. HC11, MCF10A mammary epithelial and T47D mammary carcinoma cell lines were used in this study.
Nuclear localization of IGFBP5 was observed under two conditions: fluorescent-labeled IGFBP5 added to cells with selectively permeabilized plasma but not nuclear membrane; and cells transfected with IGFBP5 expression vectors lacking the coding region for the signal peptide. By contrast, non-permeabilized cells could be stimulated to take up IGFBP5 only into intracellular vesicles outside the nucleus and this was enhanced by adding a conjugate of polylysine and transferrin, indicating an endocytotic uptake route. In addition, cells transfected with IGFBP5 containing the signal peptide secreted IGFBP5 into the medium but did not show any detectable nuclear staining.
Nuclear localization of IGFBP5 in mammary epithelial cells required the crossing of the plasma membrane, which does not appear to occur under normal cell culture conditions. Exit of IGFBP5 from endosomal vesicles into the cytosol followed by nuclear uptake was never observed. Our results indicate a minor role or no role of nuclear IGFBP5 in mediating its IGF-independent effect in the mammary epithelium and in breast cancer.
Supported by the Austrian Science Fund FWF, SFB021 'Cell proliferation and cell death in tumors'.
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