This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend 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 Binaschi, M. Articles by Capranico, G. Search for Related Content PubMed PubMed Citation Articles by Binaschi, M. Articles by Capranico, G.

Human DNA Topoisomerase II-dependent DNA Cleavage and Yeast Cell Killing by Anthracycline Analogues¹

Division of Experimental Oncology B, Istituto Nazionale per lo Studio e la Cura dei Tumori, via Venezian 1, 20133 Milan, Italy [M. B., R. F., F. Z., G. C.]; Department of Biochemistry and Genetics, The Medical School, University of Newcastle-upon-Tyne, United Kingdom [C. A. A.]; and Department of Cellular and Molecular Sciences, St. George's Hospital Medical School, University of London, United Kingdom [M. F.]

Anthracyclines are among the most clinically useful topoisomerase II poisons. A complete understanding of their molecular mechanism is thus fundamental for a rational design of novel agents. We evaluated four anthracycline analogues with respect to human topoisomerase II-dependent DNA cleaving activity, efficiency in killing yeast cells, and uptake and retention in yeast and compared the yeast system to tumor cell line models. The yeast JN394top2–4 strain was used because it has a topoisomerase II ts gene mutation: enzyme activity is much less at 30°C than at 25°C and is completely lost at 35°C. Untransformed JN394top2–4 cells were 33-fold more sensitive to idarubicin at 25°C than at 30°C, showing that topoisomerase II is the primary drug target. Overexpression of human topoisomerase II was toxic to yeast cells when the yeast enzyme was inactivated. Drug-dependent killing of yeast cells expressing low levels of the human isoenzyme at 35°C showed that the analogues spanned a 3-log range of cytotoxic potency in yeast, as they did in tumor cells. However, the compounds were much less active against the yeast strain than mammalian tumor cell lines. Drug uptake was determined and found to be altered in yeast with respect to tumor cells. Although DNA cleavage stimulated by anthracyclines roughly correlated with cytotoxicity, the cleavage level:cytotoxicity ratios were different for the studied drugs. Thus, the results suggest that other drug-dependent molecular factors contribute to drug activity in addition to the cellular content of topoisomerase II and drug uptake.

¹ This work has partially been supported by the Associazione Italiana per la Ricerca sul Cancro, Milan, Italy; the Consiglio Nazionale delle Ricerche, Progetto Finalizzato Applicazioni Cliniche della Ricerca Oncologica, Rome, Italy; and European Commission Concerted Action Contract 94-1318 on "Anticancer Drug Action on Topoisomerase II," Brussels, Belgium.

² To whom requests for reprints should be addressed, at Istituto Nazionale Tumori, 20133 Milan, Italy. Phone: +39-2-2390203; Fax: +39-2-2390764; E-mail: capranico@istitutotumori.mi.it.

Received 10/ 3/97. Accepted 3/ 3/98.

This article has been cited by other articles:

				C.-Y. Xie, H. Zhu, L.-P. Lin, Z.-H. Miao, M.-Y. Geng, Y.-J. Cai, Y. Chen, H.-J. Zhao, H.-B. Luo, X.-W. Zhang, et al. MFTZ-1, an actinomycetes subspecies derived antitumor macrolide, functions as a novel topoisomerase II poison Mol. Cancer Ther., November 1, 2007; 6(11): 3059 - 3070. [Abstract] [Full Text] [PDF]

				H.-R. Lu, H. Zhu, M. Huang, Y. Chen, Y.-J. Cai, Z.-H. Miao, J.-S. Zhang, and J. Ding Reactive Oxygen Species Elicit Apoptosis by Concurrently Disrupting Topoisomerase II and DNA-Dependent Protein Kinase Mol. Pharmacol., October 1, 2005; 68(4): 983 - 994. [Abstract] [Full Text] [PDF]

				S. Marchini, G. Damia, M. Broggini, G. Pennella, M. Ripamonti, A. Marsiglio, and C. Geroni 4-Demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyl-daunorubicin (PNU-159548), a Novel Anticancer Agent Active against Tumor CellLines with Different Resistance Mechanisms Cancer Res., March 1, 2001; 61(5): 1991 - 1995. [Abstract] [Full Text]

				M. Binaschi, M. E. Borgnetto, and G. Capranico Loss of drug-stimulated topoisomerase II DNA breaks in living cells is different at two unrelated loci Nucleic Acids Res., September 1, 2000; 28(17): 3289 - 3293. [Abstract] [Full Text] [PDF]

				M. Binaschi, R. Farinosi, M. E. Borgnetto, and G. Capranico In Vivo Site Specificity and Human Isoenzyme Selectivity of Two Topoisomerase II-poisoning Anthracyclines Cancer Res., July 1, 2000; 60(14): 3770 - 3776. [Abstract] [Full Text]

				Z. Zhou, L. A. Zwelling, Y. Kawakami, T. An, K. Kobayashi, C. Herzog, and E. S. Kleinerman Adenovirus-mediated Human Topoisomerase II{{alpha}} Gene Transfer Increases the Sensitivity of Etoposide-resistant Human Breast Cancer Cells Cancer Res., September 1, 1999; 59(18): 4618 - 4624. [Abstract] [Full Text] [PDF]

				F. Guano, P. Pourquier, S. Tinelli, M. Binaschi, M. Bigioni, F. Animati, S. Manzini, F. Zunino, G. Kohlhagen, Y. Pommier, et al. Topoisomerase Poisoning Activity of Novel Disaccharide Anthracyclines Mol. Pharmacol., July 1, 1999; 56(1): 77 - 84. [Abstract] [Full Text]