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Biochem Biophys Res Commun ; 521(3): 674-680, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31685208

RESUMO

Galectin-3 (Gal-3) is a multifunctional glycan-binding protein that participates in many pathophysiological events and has been described as a biomarker and potential therapeutic target for severe disorders, such as cancer. Several probes for Gal-3 or its ligands have been developed, however both the pathophysiological mechanisms and potential biomedical applications of Gal-3 remain not fully assessed. Molecular imaging using bioluminescent probes provides great sensitivity for in vivo and in vitro analysis for both cellular and whole multicellular organism tracking and target detection. Here, we engineered a chimeric molecule consisting of Renilla luciferase fused with mouse Gal-3 (RLuc-mGal-3). RLuc-mGal-3 preparation was highly homogenous, soluble, active, and has molecular mass of 65,870.95 Da. This molecule was able to bind to MKN45 cell surface, property which was inhibited by the reduction of Gal-3 ligands on the cell surface by the overexpression of ST6GalNAc-I. In order to obtain an efficient and stable delivery system, RLuc-mGal-3 was adsorbed to poly-lactic acid nanoparticles, which increased binding to MKN45 cells in vitro. Furthermore, bioluminescence imaging showed that RLuc-mGal-3 was able to indicate the presence of implanted tumor in mice, event drastically inhibited by the presence of lactose. This novel bioluminescent chimeric molecule offers a safe and highly sensitive alternative to fluorescent and radiolabeled probes with potential application in biomedical research for a better understanding of the distribution and fate of Gal-3 and its ligands in vitro and in vivo.


Assuntos
Galectina 3/metabolismo , Luciferases de Renilla/metabolismo , Substâncias Luminescentes/metabolismo , Neoplasias/diagnóstico por imagem , Polissacarídeos/metabolismo , Animais , Linhagem Celular Tumoral , Galectina 3/análise , Galectina 3/genética , Humanos , Luciferases de Renilla/análise , Luciferases de Renilla/genética , Substâncias Luminescentes/análise , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/metabolismo , Imagem Óptica , Polissacarídeos/análise , Ligação Proteica , Engenharia de Proteínas , Proteínas Recombinantes de Fusão/análise , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
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