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

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-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.

• Authors: Lubomir Kovar, Tomas Etrych, Martina Kabesova, Vladimir Subr, David Vetvicka, Ondrej Hovorka, Jiri Strohalm, Jan Sklenar, Petr Chytil, Karel Ulbrich, Blanka Rihova
• Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine (Tumour Biol) Vol. 31 Issue 4 Pg. 233-42 (Aug 2010) ISSN: 1423-0380 [Electronic] Netherlands
• PMID: 20556593 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Amides
• Antibiotics, Antineoplastic
• Drug Carriers
• Galectin 1
• Leukosialin
• Polymethacrylic Acids
• doxorubicin-N-(2-hydroxypropyl)methacrylamide copolymer conjugate
• Doxorubicin

Topics

• Amides (chemistry)
• Animals
• Antibiotics, Antineoplastic (pharmacology)
• Apoptosis
• Blotting, Western
• Cell Line, Tumor (drug effects)
• Cell Proliferation
• Doxorubicin (analogs & derivatives, pharmacology)
• Drug Carriers
• Endoplasmic Reticulum (metabolism)
• Flow Cytometry
• Galectin 1 (metabolism)
• Glycosylation
• Golgi Apparatus (metabolism)
• Leukosialin (metabolism)
• Lymphoma, T-Cell (drug therapy, metabolism, pathology)
• Mice
• Polymethacrylic Acids (pharmacology)