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 Deffie, A. M. Articles by Goldenberg, G. J. Search for Related Content PubMed PubMed Citation Articles by Deffie, A. M. Articles by Goldenberg, G. J.

Direct Correlation between DNA Topoisomerase II Activity and Cytotoxicity in Adriamycin-sensitive and -resistant P388 Leukemia Cell Lines¹

The Manitoba Institute of Cell Biology [A. M. D., J. K. B., G. J. G.] and the Department of Medicine, University of Manitoba [G. J. G.], Winnipeg, Manitoba, Canada R3E 0V9

The relationship between DNA topoisomerase II activity and drug resistance was studied in cloned cell lines of Adriamycin (ADR)-sensitive and -resistant P388 leukemia; drug resistant P388/ADR/3 (clone 3) and P388/ADR/7 (clone 7) cells are 5- and 10-fold more resistant to ADR than the sensitive cell line P388/4 (Cancer Res., 46: 2978, 1986). Topoisomerase II catalytic activity in crude nuclear extracts was reduced in drug-resistant cells as determined qualitatively by decatenation of kDNA. Using the centrifugal method of quantitative analysis, topoisomerase II catalytic activity (mean ± SE) was 81 ± 10 units/mg total nuclear protein in sensitive cells, 29 ± 2 units/mg total nuclear protein in resistant clone 3 cells, and 16 ± 2 units/mg total nuclear protein in resistant clone 7 cells; these differences were highly significant (P < 0.005). Similarly, quantitative analysis of DNA cleavage activity using 3' ³²P-end-labeled pBR322 restriction fragments showed that drug-stimulated topoisomerase II cleavage activity in nuclear extracts from sensitive cells was approximately 1.7- and 2.9-fold greater than that from resistant clone 3 and 7 cells, respectively. Western blot analysis of nuclear extracts from the three cell lines using antibody against the C-terminal half of recombinant-prepared human topoisomerase II polypeptide revealed reduced immunoreactivity of topoisomerase II protein in the drug-resistant cells. These data suggest that reduced topoisomerase II activity in resistant cells, which may represent quantitative reduction of the enzyme, may be another property contributing to multifactorial drug resistance in these cells.

¹ Supported by a grant from the National Cancer Institute of Canada.

² To whom requests for reprints should be addressed, at the Manitoba Institute of Cell Biology, 100 Olivia Street, Winnipeg, R3E 0V9, Canada.

Received 4/20/88. Revised 8/ 4/88. Accepted 9/29/88.

This article has been cited by other articles:

				V. Gouaze, Y.-Y. Liu, C. S. Prickett, J. Y. Yu, A. E. Giuliano, and M. C. Cabot Glucosylceramide Synthase Blockade Down-Regulates P-Glycoprotein and Resensitizes Multidrug-Resistant Breast Cancer Cells to Anticancer Drugs Cancer Res., May 1, 2005; 65(9): 3861 - 3867. [Abstract] [Full Text] [PDF]

				D. Yu, E. Khan, M. A. Khaleque, J. Lee, G. Laco, G. Kohlhagen, S. Kharbanda, Y.-C. Cheng, Y. Pommier, and A. Bharti Phosphorylation of DNA Topoisomerase I by the c-Abl Tyrosine Kinase Confers Camptothecin Sensitivity J. Biol. Chem., December 10, 2004; 279(50): 51851 - 51861. [Abstract] [Full Text] [PDF]

				B. Shiotani and H. Ashida 3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) triggers apoptosis by DNA double-strand breaks caused by inhibition of topoisomerase I Carcinogenesis, July 1, 2004; 25(7): 1149 - 1155. [Abstract] [Full Text] [PDF]

				V. Gouaze, J. Y. Yu, R. J. Bleicher, T.-Y. Han, Y.-Y. Liu, H. Wang, M. M. Gottesman, A. Bitterman, A. E. Giuliano, and M. C. Cabot Overexpression of glucosylceramide synthase and P-glycoprotein in cancer cells selected for resistance to natural product chemotherapy Mol. Cancer Ther., May 1, 2004; 3(5): 633 - 640. [Abstract] [Full Text] [PDF]

				A. Senchenkov, D. A. Litvak, and M. C. Cabot Targeting Ceramide Metabolism--a Strategy for Overcoming Drug Resistance J Natl Cancer Inst, March 7, 2001; 93(5): 347 - 357. [Abstract] [Full Text] [PDF]

				F. Kanzawa, F. Koizumi, Y. Koh, T. Nakamura, Y. Tatsumi, H. Fukumoto, N. Saijo, T. Yoshioka, and K. Nishio In Vitro Synergistic Interactions between the Cisplatin Analogue Nedaplatin and the DNA Topoisomerase I Inhibitor Irinotecan and the Mechanism of this Interaction Clin. Cancer Res., January 1, 2001; 7(1): 202 - 209. [Abstract] [Full Text]

				S. Patel, B. A. Keller, and L. M. Fisher Mutations at Arg486 and Glu571 in Human Topoisomerase IIalpha Confer Resistance to Amsacrine: Relevance for Antitumor Drug Resistance in Human Cells Mol. Pharmacol., April 1, 2000; 57(4): 784 - 791. [Abstract] [Full Text]

				Y. Mizutani, O. Yoshida, T. Miki, and B. Bonavida Synergistic Cytotoxicity and Apoptosis by Apo-2 Ligand and Adriamycin against Bladder Cancer Cells Clin. Cancer Res., September 1, 1999; 5(9): 2605 - 2612. [Abstract] [Full Text] [PDF]

				S. Kishi, N. Goto, T. Nakamura, and T. Ueda Evaluation of Cell-killing Effects of 1-{beta}-D-Arabinofuranosylcytosine and Daunorubicin by a New Computer-controlled in Vitro Pharmacokinetic Simulation System Cancer Res., June 1, 1999; 59(11): 2629 - 2634. [Abstract] [Full Text] [PDF]

				H. Komatani, M. Morita, N. Sakaizumi, K. Fukasawa, E. Yoshida, A. Okura, T. Yoshinari, and S. Nishimura A New Mechanism of Acquisition of Drug Resistance by Partial Duplication of Topoisomerase I Cancer Res., June 1, 1999; 59(11): 2701 - 2708. [Abstract] [Full Text] [PDF]

				S. Patel, A. U. Sprung, B. A. Keller, V. J. Heaton, and L. M. Fisher Identification of Yeast DNA Topoisomerase II Mutants Resistant to the Antitumor Drug Doxorubicin: Implications for the Mechanisms of Doxorubicin Action and Cytotoxicity Mol. Pharmacol., October 1, 1997; 52(4): 658 - 666. [Abstract] [Full Text]

				T. Makatsoris and M. V. Seiden High-Dose Therapy for Ovarian Carcinoma Oncologist, October 1, 1997; 2(5): 330 - 339. [Abstract] [Full Text] [PDF]