Improved immunocytochemical detection of daunomycin |
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Authors: | Koji Ohara Masashi Shin Lars-Inge Larsson Kunio Fujiwara |
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Institution: | (1) Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, Kumamoto 860-0082, Japan;(2) Division of Cell Biology, Department of Anatomy and Physiology (L-I L), The Royal Veterinary and Agricultural University, Frederiksberg, Denmark |
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Abstract: | Improved immunocytochemical (ICC) detection of the anthracycline anticancer antibiotic daunomycin (DM) has been achieved by
use of hydrogen peroxide oxidation prior to ICC staining for DM. The new method greatly enhanced the localization of DM accumulation
in cardiac, smooth and skeletal muscle of rats after a single i.v. dose of the drug. DM accumulated in the nuclei as well
as in the sarcoplasm, where it occurred in the form of small granules, which were particularly evident in cardiac muscle cells.
The distribution of the granules coincided with that of mitochondria. Uptake of DM in nuclei and mitochondria of heart muscle
cells may help to improve our understanding of the cardiac toxicity of DM and related anthracyclin antibiotics. A number of
ELISA tests were carried out in order to elucidate the mechanims of H2O2−assisted antigen retrieval. A possible mechanism is that DM is reduced and converted to its semiquinone and/or hydroquinone
derivative in vivo. Oxidation by hydrogen peroxide acts to convert these derivatives back to the native antigen. The improved
ICC methodology using oxidation to recreate native antigens from reduced metabolites may be helpful also with respect to the
localization of other drugs. |
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Keywords: | Daunomycin Immunocytochemistry Hydrogen peroxide oxidation Antigen retrieval Accumulation Heart Rat |
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