Studies on Cytotoxicity and Antioxidant activities of Lectin-like proteins from Phytoparasites (Cuscuta europaea)

Objective: Lectins are of high interest among glycoproteins, functionally vital proteins of both plant and animal cells. Lectin-like proteins (LLP) from phytoparasites are explored. The work was initiated to study cytotoxic and not much antioxidant activities of lectin-like glycoproteins isolated from the dodder ( . Cuscuta europaea) Material and methods: LLP were isolated from the seeds of the plant by extraction with phosphate buffer solution with subsequent gradient precipitation with 20% and 50% ammonium sulfate. After centrifugation the supernatants: S and S , as well 20 50 as LLP and LLP precipitations were dialyzed against the distilled water. The fractions have been established to 20 50 manifest the highest hemagglutinating activity. The carbohydrate specificity was studied in these fractions to demonstrate LLP glucose-and mannose-specificity. Cytotoxic activity of LLP was assessed biochemically in B-16 50 (skin cancer) and C127 (breast cancer) cell cultures by MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2Htetrazolium bromide) method and calculation of viable cells, and morphologically. Results: Comparative analysis of cytotoxic effect produced by various LLP fractions on B-16 and C-127 cells demonstrated the most pronounced cytotoxic effect of the proteins in B-16 cells, that melanoma B-16 cells are more sensitive to LLP Cuscuta europaea influence. S and LLP fractions of dodder showed little antioxidant properties reducing the level of MDA, and also 20 20 activating antioxidant protective enzymes -superoxide dismutase. Conclusions: Thus, we isolated and characterized LLP form as well as demonstrated cytotoxic effect of LLP and LLP on C127 and B-16 cells. For Cuscuta europaea, 20 50

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