The Rho GTPase Rac1 is a multifunctional protein working through different effector pathways. The emerging physiological significance of glycanlectin recognition gives reason to testing the possibility for an influence of modulation of Rac1 expression on these molecular aspects. Using human colon adenocarcinoma (SW620) cells genetically engineered for its up- and down-regulation (Rac1+ and Rac1- cells) along with wild-type and mock-transfected control cells, the questions are addressed whether the presence of adhesion/growth-regulatory galectins and distinct aspects of cell surface glycosylation are affected. Proceeding from RT-PCR data to Western blotting after two-dimensional gel electrophoresis and flow cytofluorimetry with non-crossreactive antibodies against six members of this lectin family (i.e. galectins-1, -3, -4, -7, -8 and -9), a reduced extent of the presence of galectins-1, -7 and -9 was revealed in the case of Rac1 cells. Application of these six galectins as probes to determination of cell reactivity for human lectins yielded relative increases in surface labelling of Rac1- cells with galectins-1, -3 and -7. Examining distinct aspects of cell surface glycosylation with a panel of 14 plant/fungal lectins disclosed a decrease in α2,6-sialylation of N-glycans and an increase in PNA-reactive sites (i.e. non-sialylated core 1 O-glycans), two alterations known to favour reactivity for galectins-1 and -3. Thus, manipulation of Rac1 expression selectively affects the expression pattern within the galectin network at the level of proteins and distinct aspects of cell surface glycosylation.
- MeSH
- 2D gelová elektroforéza MeSH
- buněčná membrána metabolismus MeSH
- fenotyp MeSH
- galektiny genetika metabolismus MeSH
- glykosylace MeSH
- kyselina N-acetylneuraminová metabolismus MeSH
- lektiny metabolismus MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory tračníku genetika metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- polysacharidy metabolismus MeSH
- průtoková cytometrie MeSH
- rac1 protein vázající GTP metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nuclear galectins participate in splicing of pre-mRNA. In this study we detected galectins-1, -2, -3 and -7 and their glycoligands in three types of cells: fibroblasts, cancer epithelial cells and melanoma cells. The results demonstrated that the nuclear expression of distinct types of galectins and their ligands in interphasic nuclei is dependent on the cell type. The extensive binding of labelled galectins-1 and -2 to mitotic cells (around chromosomes, in mitotic spindle and in bridge connecting both daughter cells) suggests their role during the cell division.
- MeSH
- buněčné jádro metabolismus MeSH
- epitelové buňky cytologie metabolismus MeSH
- fibroblasty cytologie metabolismus MeSH
- galektin 1 MeSH
- galektiny metabolismus MeSH
- interfáze fyziologie MeSH
- lidé MeSH
- ligandy MeSH
- melanocyty cytologie metabolismus MeSH
- melanom metabolismus MeSH
- mitóza fyziologie MeSH
- molekuly buněčné adheze metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory metabolismus MeSH
- růstové látky metabolismus MeSH
- spinocelulární karcinom metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The glycophenotyping of mammalian cells with plant lectins maps aspects of the glycomic profile and disease-associated alterations. A salient step toward delineating their functional dimension is the detection of endogenous lectins. They can translate sugar-encoded changes into cellular responses. Among them, the members of the lectin family of galectins are emerging regulators of cell adhesion, migration and proliferation. Focusing on galectins-1, -3 and -7, we addressed the issue whether their expression is regulated during wound healing in porcine skin as model. A conspicuous upregulation is detected for galectin-1 in the dermis and a neoexpression in the epidermis, where an increased level of galectin-7 was also found. Applying biotinylated tissue lectins as probes, the signal intensities for accessible binding sites decreased, intimating an interaction of the cell lectin with reactive sites. In contrast, galectin-3 parameters remained rather constant. Of note, epidermal cells in culture also showed an increase in expression/presence of galectin-1, measured on the levels of mRNA and protein, in this case by Western blotting and quantitative immunocytochemistry. Used as matrix, galectin-1 conferred resistance to trypsin treatment to attached human keratinocytes and reduced migration into scratch-wound areas in vitro. This report thus presents new information on endogenous lectins in wound healing and differential regulation among the three tested cases.
- MeSH
- buněčná adheze MeSH
- časové faktory MeSH
- financování organizované MeSH
- galektin 1 metabolismus MeSH
- galektin 3 metabolismus MeSH
- galektiny genetika metabolismus MeSH
- hojení ran MeSH
- imunohistochemie MeSH
- keratinocyty metabolismus patologie MeSH
- kultivované buňky MeSH
- kůže metabolismus patologie zranění MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- miniaturní prasata MeSH
- pohyb buněk MeSH
- prasata MeSH
- upregulace MeSH
- vazebná místa MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
Glycans of natural glycoconjugates are considered as a source of biological information relevant to cell adhesion or growth. Sugar-based messages are decoded and translated into responses by endogenous lectins. This mechanism assigns a functional dimension to tumour-associated changes of glycosylation. Consequently, it calls for mapping the lectin presence in tumours. Such an analysis has so far commonly been performed with the scope to determine expression of a few distinct proteins, e.g. from the effector family of galectins with focus on galectins-1 and -3. Due to the emerging evidence for functional divergence among galectins it is timely to address the challenge to evaluate their presence beyond these few family members. Having raised a panel of non-cross- -reactive antibodies against seven human galectins covering all three subfamilies, we de scribe their expression profiles in human skin. Comparison of normal and malignant tissues enabled us to define galectin-type-dependent alterations, arguing in favour of distinct functionalities. It is concluded that comprehensive monitoring performed to define the different aspects of the galectin network, as documented in this pilot study, is advisable for future histopathologic studies aimed at delineating clinical correlations.
- MeSH
- buněčná adheze MeSH
- financování organizované MeSH
- fluorescence MeSH
- galektiny metabolismus MeSH
- imunohistochemie MeSH
- kůže metabolismus MeSH
- lektiny metabolismus MeSH
- lidé MeSH
- mezibuněčné signální peptidy a proteiny metabolismus MeSH
- nádory metabolismus patologie MeSH
- zmrazené řezy MeSH
- Check Tag
- lidé MeSH
