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Doxorubicin attached to HPMA copolymer via amide bond modifies the glycosylation pattern of EL4 cells

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Tumor Biology

Abstract

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-HPMAAM, PK1) or a hydrazone pH-sensitive bond (Dox-HPMAHYD). In contrast to Dox and Dox-HPMAHYD, Dox-HPMAAM 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-HPMAAM conjugate, in contrast to Dox or Dox-HPMAHYD, 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-HPMAAM 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-HPMAAM possesses increased sensitivity to galectin-1-induced apoptosis. In this study, we demonstrate that Dox-HPMAAM treatment changes glycosylation of the EL4 T cell lymphoma surface and sensitizes the cells to galectin-1-induced apoptosis.

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Acknowledgment

This research was supported by the Grant Agency of the Czech Republic grant 305/07/P172, IAAX00500803, by the Institutional Research Concept AV0Z50200510, and by grant of the Academy of Sciences of the Czech Republic no. KAN200200651.

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Authors and Affiliations

  1. Department of Immunology and Gnotobiology, Institute of Microbiology, ASCR v.v.i, Vídenska 1083, 142 20, Prague 4, Czech Republic

    Lubomir Kovar, Martina Kabesova, David Vetvicka, Ondrej Hovorka, Jan Sklenar & Blanka Rihova

  2. Department of Biomedical Polymers, Institute of Macromolecular Chemistry, ASCR v.v.i, Heyrovskeho nam. 2, 162 06, Prague 6, Czech Republic

    Tomas Etrych, Vladimir Subr, Jiri Strohalm, Petr Chytil & Karel Ulbrich

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  1. Lubomir Kovar
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Correspondence to Lubomir Kovar.

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Kovar, L., Etrych, T., Kabesova, M. et al. Doxorubicin attached to HPMA copolymer via amide bond modifies the glycosylation pattern of EL4 cells. Tumor Biol. 31, 233–242 (2010). https://doi.org/10.1007/s13277-010-0019-7

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  • DOI: https://doi.org/10.1007/s13277-010-0019-7

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