|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Effect of nitric oxide on the daunorubicin efflux mechanism in K562 cells
Juliana Costa Curta, Ana Carolina Rabello de Moraes, Marley Aparecida Licínio, Aline Costa and Maria Cláudia Santos‑Silva1
Departamento de Anlises Clnicas, Universidade Federal de Santa Catarina, Campus Trindade, CEP 88040900, Florianpolis, SC, Brazil
NO (nitric oxide) donating drugs have been investigated for their important role in the sensitization of neoplastic cells to chemotherapy drugs. The goal of this work was to investigate the involvement of NO in the resistance of K562 cells to DNR (daunorubicin). Only simultaneous addition of DNR and SNAP (S-nitroso-N-acetyl-dl-penicillamine) caused significant cell death by apoptosis. Combination of the compounds decreased Bcl-2 and survivin, and increased Bax and active-caspase 3 expression. Fluorescence microscope and cytometric analysis showed that DNR and SNAP together caused DNR intracellular accumulation in K562 cells. RT–PCR (reverse transcription–PCR) analysis showed that DNR and SNAP, alone or in association, produced significant decreases in lrp expression. abcc1 gene expression was unaffected by the presence of SNAP, but when treated with DNR there was a small reduction that was intensified by DNR and SNAP in combination. The transport mechanism involved in the resistance to DNR in K562 cells involves ABCC1 and LRP (lung resistance protein) resistance proteins. DNR and SNAP inhibition of the expression of these proteins occurs by distinct mechanisms, and this disrupts the K562 resistance to DNR.
Key words: ABCC1, apoptosis, danorubicin, lung resistance protein (LRP), nitric oxide, multidrug resistance (MDR)
Abbreviations: AML, acute myeloid leukaemia, DAPI, 4′,6-diamidino-2-phenylindole, DNR, daunorubicin, GAPDH, glyceraldehyde-3-phosphate dehydrogenase, HRP, horseradish peroxidase, LRP, lung resistance protein, MDR, multidrug resistance, MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide, NO, oxide nitric, RT–PCR, reverse transcription–PCR, SNAP, S-nitroso-N-acetyl-dl-penicillamine
1To whom correspondence should be addressed (email firstname.lastname@example.org).
Received 7 April 2011/9 December 2011; accepted 24 February 2012
Published as Cell Biology International Immediate Publication 24 February 2012, doi:10.1042/CBI20110193
© The Author(s) Journal compilation © 2012 International Federation for Cell Biology