N-Acetyllactosamine (LacNAc; Galβ4GlcNAc) is a typical disaccharide ligand of galectins. The most abundant members of these human lectins, galectin-1 (Gal-1) and galectin-3 (Gal-3), participate in a number of pathologies including cancerogenesis and metastatic formation. In this study, we synthesized a series of fifteen N-(2-hydroxypropyl)methacrylamide (HPMA)-based glycopolymers with varying LacNAc amounts and presentations and evaluated the impact of their architecture on the binding affinity to Gal-1 and Gal-3. The controlled radical reversible addition-fragmentation chain transfer copolymerization technique afforded linear polymer precursors with comparable molecular weight (Mn ≈ 22,000 g mol-1) and narrow dispersity (D̵ ≈ 1.1). The precursors were conjugated with the functionalized LacNAc disaccharide (4-22 mol % content in glycopolymer) prepared by enzymatic synthesis under catalysis by β-galactosidase from Bacillus circulans. The structure-affinity relationship study based on the enzyme-linked immunosorbent assay revealed that the type of LacNAc presentation, individual or clustered on bi- or trivalent linkers, brings a clear discrimination (almost 300-fold) between Gal-1 and Gal-3, reaching avidity to Gal-1 in the nanomolar range. Whereas Gal-1 strongly preferred a dense presentation of individually distributed LacNAc epitopes, Gal-3 preferred a clustered LacNAc presentation. Such a strong galectin preference based just on the structure of a multivalent glycopolymer type is exceptional. The prepared nontoxic, nonimmunogenic, and biocompatible glycopolymers are prospective for therapeutic applications requiring selectivity for one particular galectin.
- MeSH
- akrylamidy chemie MeSH
- aminocukry chemie MeSH
- Bacillus enzymologie MeSH
- beta-galaktosidasa metabolismus MeSH
- disacharidy chemická syntéza MeSH
- ELISA MeSH
- epitopy MeSH
- galektin 1 analýza metabolismus MeSH
- galektiny analýza metabolismus MeSH
- katalýza MeSH
- krevní proteiny analýza metabolismus MeSH
- magnetická rezonanční spektroskopie MeSH
- polymerizace MeSH
- polymery chemie metabolismus farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND/AIM: Having previously initiated genome-wide expression profiling in head and neck squamous cell carcinoma (HNSCC) for regions of the tumor, the margin of surgical resecate (MSR) and normal mucosa (NM), we here proceed with respective analysis of cases after stratification according to the expression status of tenascin (Ten). MATERIALS AND METHODS: Tissue specimens of each anatomical site were analyzed by immunofluorescent detection of Ten, fibronectin (Fn) and galectin-1 (Gal-1) as well as by microarrays. RESULTS: Histopathological examination demonstrated that Ten+Fn+Gal-1+co-expression occurs more frequently in samples of HNSCC (55%) than in NM (9%; p<0.01). Contrary, the Ten-Fn+Gal-1-(45%) and Ten-Fn-Gal-1-(39%) status occurred with significantly (p<0.01) higher frequency than in HNSCC (3% and 4%, respectively). In MSRs, different immunophenotypes were distributed rather equally (Ten+Fn+Gal-1+=24%; Ten-Fn+Gal-1-=36%; Ten-Fn-Gal-1-=33%), differing to the results in tumors (p<0.05). Absence/presence of Ten was used for stratification of patients into cohorts without a difference in prognosis, to comparatively examine gene-activity signatures. Microarray analysis revealed i) expression of several tumor progression-associated genes in Ten+HNSCC tumors and ii) a strong up-regulation of gene expression assigned to lipid metabolism in MSRs of Ten-tumors, while NM profiles remained similar. CONCLUSION: The presented data reveal marked and specific changes in tumors and MSR specimens of HNSCC without a separation based on prognosis.
- MeSH
- fibronektiny genetika metabolismus MeSH
- galektin 1 genetika metabolismus MeSH
- genová ontologie MeSH
- lidé MeSH
- nádorové biomarkery genetika MeSH
- nádory hlavy a krku genetika metabolismus chirurgie MeSH
- přežití po terapii bez příznaků nemoci MeSH
- regulace genové exprese u nádorů * MeSH
- resekční okraje MeSH
- sliznice metabolismus MeSH
- spinocelulární karcinom genetika metabolismus chirurgie MeSH
- stanovení celkové genové exprese metody MeSH
- tenascin genetika metabolismus MeSH
- transkriptom * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Galektíny patria medzi endogénne lektíny – bielkoviny špecificky rozpoznávajúce cukorné motívy. Galektíny hrajú významné úlohy v procesoch bunkovej proliferácie, diferenciácie, migrácie a tvorby medzibunkovej hmoty. Navyše sú schopné prenášať bunkové signály a podieľať sa na medzibunkových interakciách. Podobne bolo dokázané, že galektíny zohrávajú dôležitú úlohu pri tvorbe mikroprostredia nádoru a/alebo hojacej sa rany. Táto práca poskytuje prehľad experimentálnych a klinických štúdií zaoberajúcich sa biologickými úlohami galektínov v tkanivovej reparácii a jej paralele k raste nádoru.
Galectins are representatives of endogenous lectins – molecules specifically recognizing distinct sugar motifs. They play an important role in the processes of cell proliferation, differentiation, migration and extracellular matrix formation. Furthermore, galectins are able to transfer cellular signals and to participate in intercellular interaction. It has been proven that galectins play an important role in the formation of tumor and/or wound healing microenvironment. This review contains an overview of experimental and clinical studies dealing with biological roles of galectins in tissue repair and in its parallel – the tumor growth.
- MeSH
- exprese genu fyziologie MeSH
- galektin 1 fyziologie metabolismus MeSH
- galektin 3 fyziologie metabolismus MeSH
- galektiny * fyziologie klasifikace metabolismus MeSH
- hojení ran fyziologie MeSH
- lidé MeSH
- modely u zvířat MeSH
- nádory patofyziologie MeSH
- tkáně fyziologie metabolismus MeSH
- výzkum MeSH
- zánět patofyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- přehledy MeSH
Cancer-associated fibroblasts (CAFs) play a role in the progression of malignant tumors. They are formed by conversion of fibroblasts to smooth muscle α-actin-positive (SMA-positive) myofibroblasts. Polyamines are known to change the arrangement of the actin cytoskeleton by binding to the anionic actin. We tested the effect of the synthetic polyamine BPA-C8 on the transition of human dermal fibroblasts to myofibroblasts induced either by TGF-β1 alone or by TGF-β1 together with adhesion/growth-regulatory galectin-1. Pre-existing CAFs, myofibroblasts from pancreatitis, and rat smooth muscle cells were also exposed to BPA-C8. BPA-C8 impaired myofibroblast formation from activated fibroblasts, but it had no effect on cells already expressing SMA. BPA-C8 also reduced the occurrence of an extracellular matrix around the activated fibroblasts. The reported data thus extend current insights into polyamine activity, adding interference with tumor progression to the tumor-promoting processes warranting study.
- MeSH
- aktiny metabolismus MeSH
- fibroblasty účinky léků patologie MeSH
- galektin 1 metabolismus MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- myofibroblasty účinky léků patologie MeSH
- nádorové buňky kultivované MeSH
- nádory farmakoterapie metabolismus patologie MeSH
- polyaminy chemie farmakologie MeSH
- škára cytologie účinky léků MeSH
- transformující růstový faktor beta1 metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
INTRODUCTION: One route of translating the information encoded in the glycan chains of cellular glycoconjugates into physiological effects is via receptor (lectin) binding. A family of endogenous lectins, sharing folding, a distinct sequence signature and affinity for β-galactosides (thus termed galectins), does so effectively in a context-dependent manner. AREAS COVERED: An overview is given on the multifunctional nature of galectins, with emphasis on galectin-1. The broad range of functions includes vital processes such as adhesion via glycan bridging, glycoconjugate transport or triggering signaling relevant, for example, for growth regulation. Besides distinct glycoconjugates, this lectin can also interact with certain proteins so that it can target counterreceptors at all sites of location, that is, in the cytoplasm and/or nucleus, at both sides of the membrane or extracellularly. Approaches to strategically exploit galectin activities with therapeutic intentions are outlined. EXPERT OPINION: The wide versatility of sugar coding and the multifunctionality of galectin-1 explain why considering to turn the protein into a therapeutic target is an ambitious aim. Natural pathways shaped by physiologic master regulators (e.g., the tumor suppressor p16(INK4a)) are suggested to teach inspiring lessons as to how the lectin might be recruited to clinical service.
- MeSH
- antitumorózní látky farmakologie terapeutické užití MeSH
- biologický transport účinky léků MeSH
- buněčná adheze účinky léků MeSH
- cílená molekulární terapie MeSH
- galektin 1 antagonisté a inhibitory metabolismus MeSH
- galektiny antagonisté a inhibitory metabolismus MeSH
- glykokonjugáty metabolismus MeSH
- lidé MeSH
- nádorové proteiny antagonisté a inhibitory metabolismus MeSH
- nádory farmakoterapie prevence a kontrola MeSH
- proliferace buněk účinky léků MeSH
- racionální návrh léčiv * MeSH
- signální transdukce účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Previously, we found that treatment of cutaneous wounds with Atropa belladonna L. (AB) revealed shortened process of acute inflammation as well as increased tensile strength and collagen deposition in healing skin wounds (Gál et al. 2009). To better understand AB effect on skin wound healing male Sprague-Dawley rats were submitted to one round full thickness skin wound on the back. In two experimental groups two different concentrations of AB extract were daily applied whereas the control group remained untreated. For histological evaluation samples were removed on day 21 after surgery and stained for wide spectrum cytokeratin, collagen III, fibronectin, galectin-1, and vimentin. In addition, in the in vitro study different concentration of AB extract were used to evaluate differences in HaCaT keratinocytes proliferation and differentiation by detection of Ki67 and keratin-19 expressions. Furthermore, to assess ECM formation of human dermal fibroblasts on the in vitro level fibronectin and galectin-1 were visualized. Our study showed that AB induces fibronectin and galectin-1 rich ECM formation in vitro and in vivo. In addition, the proliferation of keratinocytes was also increased. In conclusion, AB is an effective modulator of skin wound healing. Nevertheless, further research is needed to find optimal therapeutic concentration and exact underlying mechanism of action.
- MeSH
- Atropa belladonna chemie MeSH
- časové faktory MeSH
- extracelulární matrix metabolismus účinky léků MeSH
- fibroblasty metabolismus patologie účinky léků MeSH
- fibronektiny metabolismus MeSH
- galektin 1 metabolismus MeSH
- hojení ran účinky léků MeSH
- keratin-19 metabolismus MeSH
- keratinocyty metabolismus patologie účinky léků MeSH
- kolagen typ III metabolismus MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- kůže metabolismus patologie účinky léků zranění MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- penetrující rány farmakoterapie metabolismus patologie MeSH
- potkani Sprague-Dawley MeSH
- rostlinné extrakty farmakologie chemie izolace a purifikace MeSH
- rozpouštědla chemie MeSH
- vimentin metabolismus MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tumor stroma is an active part influencing the biological properties of malignancies via molecular cross-talk. Cancer-associated fibroblasts play a significant role in this interaction. These cells frequently express smooth muscle actin and can be classified as myofibroblasts. The adhesion/growth-regulatory lectin galectin-1 is an effector for their generation. In our study, we set the presence of smooth muscle actin-positive cancer-associated fibroblasts in relation to this endogenous lectin and an in vivo competitor (galectin-3). In squamous cell carcinomas of head and neck, upregulation of galectin-1 presence was highly significantly correlated to presence of smooth muscle actin-positive cancer-associated fibroblasts in the tumor (p = 4 × 10(-8)). To pinpoint further correlations on the molecular level, we applied microarray analyses to the transcription profiles of the corresponding tumors. Significant correlations of several transcripts were detected with the protein level of galectin-1 in the cancer-associated fibroblasts. These activated genes (MAP3K2, TRIM23, PTPLAD1, FUSIP1, SLC25A40 and SPIN1) are related to known squamous-cell-carcinoma poor-prognosis factors, NF-κB upregulation and splicing downregulation. These results provide new insights into the significance of presence of myofibroblasts in squamous cell carcinoma.
- MeSH
- aktiny biosyntéza genetika metabolismus MeSH
- buňky stromatu metabolismus patologie MeSH
- down regulace MeSH
- galektin 1 biosyntéza genetika metabolismus MeSH
- galektin 3 genetika metabolismus MeSH
- genetická transkripce MeSH
- hladké svalstvo metabolismus patologie MeSH
- lidé MeSH
- myofibroblasty metabolismus patologie MeSH
- nádory hlavy a krku genetika metabolismus patologie MeSH
- NF-kappa B genetika metabolismus MeSH
- prognóza MeSH
- sestřih RNA MeSH
- spinocelulární karcinom genetika metabolismus patologie MeSH
- upregulace MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Members of the galectin family of endogenous lectins are potent adhesion/growth-regulatory effectors. Their multifunctionality opens possibilities for their use in bioapplications. We studied whether human galectins induce the conversion of human dermal fibroblasts into myofibroblasts (MFBs) and the production of a bioactive extracellular matrix scaffold is suitable for cell culture. Testing a panel of galectins of all three subgroups, including natural and engineered variants, we detected activity for the proto-type galectin-1 and galectin-7, the chimera-type galectin-3 and the tandem-repeat-type galectin-4. The activity of galectin-1 required the integrity of the carbohydrate recognition domain. It was independent of the presence of TGF-β1, but it yielded an additive effect. The resulting MFBs, relevant, for example, for tumor progression, generated a matrix scaffold rich in fibronectin and galectin-1 that supported keratinocyte culture without feeder cells. Of note, keratinocytes cultured on this substratum presented a stem-like cell phenotype with small size and keratin-19 expression. In vivo in rats, galectin-1 had a positive effect on skin wound closure 21 days after surgery. In conclusion, we describe the differential potential of certain human galectins to induce the conversion of dermal fibroblasts into MFBs and the generation of a bioactive cell culture substratum.
- MeSH
- extracelulární matrix metabolismus MeSH
- fibroblasty metabolismus MeSH
- galektin 1 metabolismus MeSH
- galektin 3 metabolismus MeSH
- galektin 4 metabolismus MeSH
- galektiny metabolismus MeSH
- hojení ran MeSH
- keratin-19 metabolismus MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- myofibroblasty metabolismus MeSH
- tkáňové inženýrství metody MeSH
- transformující růstový faktor beta1 metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
To avoid the side effects of the anti-cancer drug doxorubicin (Dox), we conjugated this drug to a N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer backbone. Dox was conjugated via an amide bond (Dox-HPMA(AM), PK1) or a hydrazone pH-sensitive bond (Dox-HPMA(HYD)). In contrast to Dox and Dox-HPMA(HYD), Dox-HPMA(AM) accumulates within the cell's intracellular membranes, including those of the Golgi complex and endoplasmic reticulum, both involved in protein glycosylation. Flow cytometry was used to determine lectin binding and cell death, immunoblot to characterize the presence of CD7, CD43, CD44, and CD45, and high-performance anion exchange chromatography with pulsed amperometric detector analysis for characterization of plasma membrane saccharide composition. Incubation of EL4 cells with Dox-HPMA(AM) conjugate, in contrast to Dox or Dox-HPMA(HYD), increased the amounts of membrane surface-associated glycoproteins, as well as saccharide moieties recognized by peanut agglutinin, Erythrina cristagalli, or galectin-1 lectins. Only Dox-HPMA(AM) increased expression of the highly glycosylated membrane glycoprotein CD43, while expression of others (CD7, CD44, and CD45) was unaffected. The binding sites for galectin-1 are present on CD43 molecule. Furthermore, we present that EL4 treated with Dox-HPMA(AM) possesses increased sensitivity to galectin-1-induced apoptosis. In this study, we demonstrate that Dox-HPMA(AM) treatment changes glycosylation of the EL4 T cell lymphoma surface and sensitizes the cells to galectin-1-induced apoptosis.
- MeSH
- amidy chemie MeSH
- antibiotika antitumorózní farmakologie MeSH
- antigeny CD43 metabolismus MeSH
- apoptóza MeSH
- doxorubicin analogy a deriváty farmakologie MeSH
- endoplazmatické retikulum metabolismus MeSH
- galektin 1 metabolismus MeSH
- glykosylace MeSH
- Golgiho aparát metabolismus MeSH
- kyseliny polymethakrylové farmakologie MeSH
- lymfom T-buněčný farmakoterapie metabolismus patologie MeSH
- myši MeSH
- nádorové buněčné linie účinky léků MeSH
- nosiče léků MeSH
- proliferace buněk MeSH
- průtoková cytometrie MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Benign and malignant fibrous histiocytoma present with a considerable difference concerning cellular organization in their vicinity. OBJECTIVE: Normally appearing epithelium covers the malignant form in contrast to hyperplastic epidermis for benign tumors. It is an open question as to whether the tumor-associated fibroblasts are capable to affect phenotypic features of normal keratinocytes, prompting this comparative analysis. METHODS: Fibroblasts were isolated from benign and malignant fibrous histiocytomas, respectively, and also from normal dermis. The resulting cell populations were thoroughly characterized immunocytochemically using a large panel of antibodies. The three fibroblast preparations were cocultured with normal interfollicular keratinocytes. Their phenotype was characterized for distinct properties including differentiation and proliferation. RESULTS: Fibroblasts prepared from both tumor types were phenotypically practically identical with normal dermal fibroblasts. Their activities on keratinocytes were different. Cells prepared from benign fibrous histiocytoma were capable to effect strong expression of keratin 19 and production of a galectin-1-rich extracellular matrix. Fibroblasts isolated from malignant fibrous histiocytoma led to a phenotype very similar to that when keratinocytes were cocultured with normal dermal fibroblasts. CONCLUSION: Fibroblasts prepared from benign fibrous histiocytoma were biologically active on keratinocytes in a particular manner. Our results on fibroblast activity are suggested to be relevant for morphologic differences observed in vivo between normal epidermis and epidermis adjacent to the studied tumor types.
- MeSH
- benigní fibrózní histiocytom metabolismus patologie MeSH
- biologické markery metabolismus MeSH
- epidermis metabolismus patologie MeSH
- fibroblasty metabolismus patologie MeSH
- galektin 1 metabolismus MeSH
- keratin-19 metabolismus MeSH
- keratinocyty metabolismus patologie MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- lidé MeSH
- maligní fibrózní histiocytom metabolismus patologie MeSH
- nádory kůže metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
