ABSTRACT
We made fusion protein of fastatin and FIII 9-10, termed tetra-cell adhesion molecule (T-CAM) that can interact simultaneously with alphavbeta3 and alpha5beta1 integrins, both playing important roles in tumor angiogenesis. T-CAM can serve as a cell adhesion substrate mediating adhesion and migration of endothelial cells in alphavbeta3 and alpha5beta1 integrin-dependent manner. T-CAM showed pronounced anti-angiogenic activities such as inhibition of endothelial cell tube formation, endothelial cell proliferation, and induction of endothelial cell apoptosis. T-CAM also inhibited angiogenesis and tumor growth in mouse xenograft model. The anti-angiogenic and anti-tumoral activity of molecule like fastatin could be improved by fusing it with integrin-recognizing cell adhesion domain from other distinct proteins. The strategy of combining two distinct anti-angiogenic molecules or cell adhesion domains could facilitate designing improved anticancer agent of therapeutic value.
Subject(s)
Animals , Humans , Male , Mice , Angiogenesis Inhibitors/chemistry , Antineoplastic Agents/chemistry , Base Sequence , Benzocaine/chemistry , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cells, Cultured , Chloramphenicol/chemistry , DNA Primers , Drug Combinations , Factor VIII/chemistry , Integrin alpha5beta1/physiology , Integrin alphaVbeta3/physiology , Mice, Inbred BALB C , Nitrofurazone/chemistry , Recombinant Fusion Proteins/chemistryABSTRACT
Uma metodologia analítica par determinaão de resíduos de clorafenicol em tecido muscular foi padronizada e validada empregando extração com acetado de etila, purificação com cartuchos de sílica e detecção e quantificação por CLAE/UV. A faixa de linearidade da resposta, sem inteferência da matriz, foi de 10 a 400 ng/mL. Nos ensaios com amostras fortificadas entre 5 e 40µg/kg