This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sinha, B. K. Articles by Myers, C. E. Search for Related Content PubMed PubMed Citation Articles by Sinha, B. K. Articles by Myers, C. E.

Adriamycin Activation and Oxygen Free Radical Formation in Human Breast Tumor Cells: Protective Role of Glutathione Peroxidase in Adriamycin Resistance¹

Biochemical Pharmacology Section, Clinical Pharmacology and Medicine Branch, Clinical Oncology Program, Division of Cancer Treatment, National Cancer Institute, NIH, Bethesda, Maryland 20892

Previous studies with Adriamycin-sensitive and -resistant (ADR^R) MCF-7 human breast tumor cell lines indicated that Adriamycin formed significantly less hydroxyl radical (·OH) as the result of enhanced detoxification of reactive oxygen intermediates in the ADR^R cell line. In order to further define the sites of drug activation and the role of detoxification mechanisms in free radical levels, subcellular fractions were isolated from these two cell lines and free radical formation in the presence of Adriamycin was examined by using electron spin resonance spectroscopy. Studies reported here show that considerable NADPH-cytochrome P-450 reductase and NADH dehydrogenase activities were present in microsomes and mitochondria, respectively, and in nuclei obtained from these cells, and the relative activity of NADH dehydrogenase was 2-fold higher in the mitochondrial fraction of ADR^R cells compared to the mitochondrial fraction from the parental wild type cells. In the presence of Adriamycin and a reducing cofactor (NADPH or NADH), Adriamycin semiquinone free radical, superoxide anion, and ·OH were detected in all these fractions. Although only a small difference in the relative amount of oxy radical formation was detected in tumor microsomes, both mitochondria and nuclei of ADR^R cells showed an overall 2-fold decreased formation of oxy radicals. The formation of the free radicals was significantly inhibited by superoxide dismutase, catalase, and dimethyl sulfoxide, indicating that free ·OH generation was both superoxide and hydrogen peroxide dependent. The addition of purified glutathione peroxidase likewise inhibited ·OH formation in a dose-dependent fashion. Similarly, when the lysate from ADR^R cells, which contains 12- to 14-fold more glutathione peroxidase than Adriamycin-sensitive cells, was added to reaction mixtures containing Adriamycin-sensitive cells and Adriamycin, the ·OH formation was diminished. Decreased free radical formation in nuclei and mitochondria, as a result of detoxification of hydrogen peroxide by glutathione peroxidase, may be significant in the protection of ADR^R cells from Adriamycin-induced cell killing.

¹ We dedicate this paper to the memory of Lata Dusre, a fellow investigator who died of cancer April 26, 1989.

² To whom requests for reprints should be addressed, at Building 10, Room 6N-119, National Cancer Institute, NIH, Bethesda, MD 20892.

Received 11/ 7/88. Revised 3/ 1/89. Accepted 4/14/89.

This article has been cited by other articles:

				C. Andreadis, P. A. Gimotty, P. Wahl, R. Hammond, J. Houldsworth, S. J. Schuster, and T. R. Rebbeck Members of the glutathione and ABC-transporter families are associated with clinical outcome in patients with diffuse large B-cell lymphoma Blood, April 15, 2007; 109(8): 3409 - 3416. [Abstract] [Full Text] [PDF]

				S. Wang, E. A. Konorev, S. Kotamraju, J. Joseph, S. Kalivendi, and B. Kalyanaraman Doxorubicin Induces Apoptosis in Normal and Tumor Cells via Distinctly Different Mechanisms: INTERMEDIACY OF H2O2- AND p53-DEPENDENT PATHWAYS J. Biol. Chem., June 11, 2004; 279(24): 25535 - 25543. [Abstract] [Full Text] [PDF]

				J. L. Brumaghim, Y. Li, E. Henle, and S. Linn Effects of Hydrogen Peroxide upon Nicotinamide Nucleotide Metabolism in Escherichia coli: CHANGES IN ENZYME LEVELS AND NICOTINAMIDE NUCLEOTIDE POOLS AND STUDIES OF THE OXIDATION OF NAD(P)H BY Fe(III) J. Biol. Chem., October 24, 2003; 278(43): 42495 - 42504. [Abstract] [Full Text] [PDF]

				S. Ramji, C. Lee, T. Inaba, A. V. Patterson, and D. S. Riddick Human NADPH-Cytochrome P450 Reductase Overexpression Does Not Enhance the Aerobic Cytotoxicity of Doxorubicin in Human Breast Cancer Cell Lines Cancer Res., October 15, 2003; 63(20): 6914 - 6919. [Abstract] [Full Text] [PDF]

				G. S. Watts, B. W. Futscher, R. Isett, M. Gleason-Guzman, M. W. Kunkel, and S. E. Salmon cDNA Microarray Analysis of Multidrug Resistance: Doxorubicin Selection Produces Multiple Defects in Apoptosis Signaling Pathways J. Pharmacol. Exp. Ther., November 1, 2001; 299(2): 434 - 441. [Abstract] [Full Text] [PDF]