Nejvíce citovaný článek - PubMed ID 14630391
New aspects of galectin functionality in nuclei of cultured bone marrow stromal and epidermal cells: biotinylated galectins as tool to detect specific binding sites
Cellular activities in the regulation of growth or adhesion/migration involve protein (lectin)-carbohydrate recognition at the cell surface. Members of the galectin family of endogenous lectins additionally bind distinct intracellular ligands. These interactions with protein targets explain the relevance of their nuclear and cytoplasmic presence. Expression profiling for galectins and accessible binding sites is a histochemical approach to link localization with cellular growth properties. Non-cross-reactive antibodies for the homodimeric (proto-type) galectins-1, -2 and -7 and the chimera-type galectin-3 (Gal-3) as well as the biotinylated lectins were tested. This analysis was performed with the FaDu squamous carcinoma cell line and long-term cultured human and porcine epidermal cells as models for malignant and normal cells of squamous cell epithelial origin. A set of antibodies was added for phenotypic cell characterization. Strong nuclear and cytoplasmic signals of galectins and the differential reactivity of labeled galectins support the notion of their individual properties. The length of the period of culture was effective in modulating marker expression. Cytochemical expression profiling is a prerequisite for the selection of distinct proteins for targeted modulation of gene expression as a step toward functional analysis.
- MeSH
- buněčná diferenciace fyziologie MeSH
- buněčné jádro metabolismus ultrastruktura MeSH
- epitelové buňky metabolismus MeSH
- fenotyp MeSH
- galektiny metabolismus MeSH
- lidé MeSH
- molekuly buněčné adheze metabolismus MeSH
- prasata MeSH
- růstové látky metabolismus MeSH
- sacharidy fyziologie MeSH
- signální transdukce fyziologie MeSH
- spinocelulární karcinom metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- galektiny MeSH
- molekuly buněčné adheze MeSH
- růstové látky MeSH
- sacharidy MeSH
The principal pool of epidermal stem cells is located in the bulge region of the hair follicle root sheath. In this research project, we have used a refined procedure to isolate porcine hair follicles including their root sheath and for comparison purposes also human cell material. These cells migrating from the hair follicles were then cytochemically characterized. A panel of antibodies and two labeled plant lectins were tested on cell material obtained under a range of assorted experimental conditions. Due to their role in growth regulation we also studied two endogenous lectins, specifically monitoring their expression and the presence of accessible ligands. These in vitro results were compared with findings on porcine and human hair follicles and human basal cell carcinomas in situ. The keratinocytes originating from hair follicles in the presence of feeder cells are rather undifferentiated and express galectin-1/galectin-1-binding sites but not galectin-3 in their nuclei associated with DeltaNp63alpha positivity. Nuclear reactivity for galectin-1 was rarely observed in the bulge of the outer root sheath of the human hair follicle and of basal cell carcinomas and absent in porcine tissue samples. Exclusion of feeder cells from our cultivation system of porcine hair follicles led to the formation of spheroid bodies from these keratinocytes. Ki67 as a marker of proliferation was not present in the nuclei of cells forming these spheroids. One part of these bodies is positive for markers of post-mitotic differentiated cells, while the other spheroids are composed of poorly differentiated cells, which are able to adhere to feeder cells and form growing colonies. In summary, the detection of galectin-1 and also nuclear binding sites for this endogenous effector points to intracellular functionality of this lectin. It can be considered a potential marker of a distinct cell population, probably at the beginning of a differentiation cascade of keratinocytes.
- MeSH
- buněčná diferenciace MeSH
- fenotyp MeSH
- galektiny analýza MeSH
- imunodominantní epitopy analýza MeSH
- keratinocyty chemie MeSH
- lidé MeSH
- prasata MeSH
- vlasový folikul chemie cytologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- galektiny MeSH
- imunodominantní epitopy MeSH