Enhancing anti-tumor efficacy of Doxorubicin by non-covalent conjugation to gold nanoparticles - in vitro studies on feline fibrosarcoma cell lines.
BACKGROUND
Feline injection-site sarcomas are malignant skin tumors of mesenchymal origin, the treatment of which is a challenge for veterinary practitioners. Methods of treatment include radical surgery, radiotherapy and chemotherapy. The most commonly used cytostatic drugs are cyclophosphamide, doxorubicin and vincristine. However, the use of cytostatics as adjunctive treatment is limited due to their adverse side-effects, low biodistribution after intravenous administration and multidrug resistance. Colloid gold nanoparticles are promising drug delivery systems to overcome multidrug resistance, which is a main cause of ineffective chemotherapy treatment. The use of colloid gold nanoparticles as building blocks for drug delivery systems is preferred due to ease of surface functionalization with various molecules, chemical stability and their low toxicity.


METHODS
Stability and structure of the glutathione-stabilized gold nanoparticles non-covalently modified with doxorubicin (Au-GSH-Dox) was confirmed using XPS, TEM, FT-IR, SAXRD and SAXS analyses. MTT assay, Annexin V and Propidium Iodide Apoptosis assay and Rhodamine 123 and Verapamil assay were performed on 4 feline fibrosarcoma cell lines (FFS1WAW, FFS1, FFS3, FFS5). Statistical analyses were performed using Graph Pad Prism 5.0 (USA).


RESULTS
A novel approach, glutathione-stabilized gold nanoparticles (4.3 +/- 1.1 nm in diameter) non-covalently modified with doxorubicin (Au-GSH-Dox) was designed and synthesized. A higher cytotoxic effect (p<0.01) of Au-GSH-Dox than that of free doxorubicin has been observed in 3 (FFS1, FFS3, FFS1WAW) out of 4 feline fibrosarcoma cell lines. The effect has been correlated to the activity of glycoprotein P (main efflux pump responsible for multidrug resistance).


CONCLUSIONS
The results indicate that Au-GSH-Dox may be a potent new therapeutic agent to increase the efficacy of the drug by overcoming the resistance to doxorubicin in feline fibrosarcoma cell lines. Moreover, as doxorubicin is non-covalently attached to glutathione coated nanoparticles the synthesized system is potentially suitable to a wealth of different drug molecules.
Faculty of Chemistry, University of Warsaw, Warsaw, Poland.
Michał
M
Wójcik
Faculty of Chemistry, University of Warsaw, Warsaw, Poland.
Wiktor
W
Lewandowski
Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Magdalena
M
Król
Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Karol
K
Pawłowski
Faculty of Chemistry, University of Warsaw, Warsaw, Poland.
Józef
J
Mieczkowski
Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Roman
R
Lechowski
Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Katarzyna
K
Zabielska
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage* / Q000737:chemistry; D017209:Apoptosis / Q000187:drug effects; D002415:Cats; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004317:Doxorubicin / Q000008:administration & dosage* / Q000737:chemistry; D005354:Fibrosarcoma; D005978:Glutathione / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D058726:Nanoconjugates / Q000737:chemistry / Q000648:ultrastructure
D000903:Antibiotics, Antineoplastic; D058726:Nanoconjugates; D006046:Gold; D004317:Doxorubicin; D005978:Glutathione