Abstract
Human ovarian cancer cells A2780, selected for resistance to doxorubicin (A2780-DX3), are crossresistant to various other topoisomerase-II-targeted drugs but not to vinblastine. The parental cell line was very sensitive to doxorubicin-, mitoxantrone- or etoposide (VP16)-induced DNA single-strand breaks, under deproteinizing conditions. In contrast, little or no DNA strand breakage was seen in resistant A2780-DX3 cells, even at very high concentrations, indicating a good correlation, with cytotoxicity. No significant alterations in cellular drug uptake were observed in DX3 cells. Further studies showed that the nuclei isolated from resistant cells were also resistant to mitoxantroneor VP16-induced single-strand breaks, indicating that nuclear modifications in resistant cells are responsible for this resistance. Catalytic activity in crude nuclear extracts from wild-type and DX3 cells was almost equal. However, an assay that specifically measures generation of 5′-protein-linked breaks in32P-labeled 3 DNA revealed that, DNA cleavage activity in nuclear extract from the DX3 cell line is profoundly resistant to a stimulation by VP16. These data indicate that stimulation of topoisomerase-II-mediated DNA cleavage is responsible for topoisomerase-II-targeted drugcytotoxicity rather than loss of normal topoisomerase catalytic function. These data support the hypothesis that A2780-DX3 cells display an “atypical” multidrug resistance.
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Abbreviations
- MDR :
-
multidrug resistance
- SSB :
-
Single-strand break
References
Beck WT (1990) Mechanisms of multidrug resistance in human tumor cells. The roles of P-glycoprotein, DNA topoisomerase II, and other factors. Cancer Treat Rev 17 [Suppl A]: 11–20
Bourhis J Goldstein, Riou G, Pastan I, Gottesman M, Benard J (1989) Expression of a human multidrug resistance gene in ovarian carcinomas. Cancer Res 49:5062–5065
Bradford MA (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing principles of protein-dye binding. Anal Biochem 72:248–254
Cimoli G, Valenti M, Parodi S, Mazzoni A, De Sessa F, Conte PF, Russo P (1993) Reversal of “atypical”-multidrug resistance by recombinant human tumor necrosis factor in the human ovarian cancer cell line DX3. Oncol Res 8:311–323
Conte PF, Chiara S, Bruzzone M, Repetto L, Guido T, Mammoliti S, Vidali A (1989) Reassessment of the role of II-look laparatomy in advanced ovarian cancer. 5th European Conference on Clinical Oncology, London, p. 1064.
Conte PF, Bruzzone, M, Chiara S, Rubagotti A, Rosso R (1990) The role of doxorubicin in the treatment of epithelial ovarian carcinoma. Proceedings of the Perugia International Cancer Conference. LP Communication, New York, pp 70–76
Drake FH, Zimmerman JP, McCabe FL, Bartus HF, Per SR, Sullivan DM, Ross WE, Mattern MR, Johnson RK, Crooke ST, Mirabelli CK (1987) Purification of topoisomerase II from amsacrine-resistant p388 leukemia cells. J Biol Chem 262:16739–16747
Drake FH, Hofman GA, Bartus HF, Mattern MR, Crooke ST, Mirabelli CK (1989) Biochemical and pharmacological properties of p170 and p180 forms of topoisomerase II. Biochemistry 28:8154–8160
Fox ME, Smith PJ (1990) Long-term inhibition of DNA synthesis and the persistence of trapped topoisomerase II complexes in determining the toxicity of the antitumor DNA intercalators mAMSA and mitoxantrone. Cancer Res 50:5813–5818
Glisson BS, Gupta R, Smallwood-Kentro S, Ross WE (1986) Characterization of acquired epipodophyllotoxin resistance in a Chinese hamster ovary cell line: loss of drug-stimulated DNA cleavage activity. Cancer Res 46:1934–1938
Glisson BS, Killary AM, Merta P, Ross WE, Siciliano J, Siciliano MJ (1992) Dissociation of cytotoxicity and DNA cleavage activity induced by topoisomerase II-reactive intercalating agents in hamster-human somatic cell hybrids. Cancer Chemother Pharmacol 31:131–138
Gruppo Interregionale Cooperativo Oncologia Ginecologica (GICOG) (1987) Randomized comparison of cisplatin with cyclophosphamide-cisplatin with cyclophosphamide-doxorubicincisplatin in advanced ovarian cancer. Lancet II:353–359
Hochhauser D, Harris AL (1993) The role of topoisomerase II and β in drug resistance. Cancer Treat Rev 19:181–194
Jong S de, Kooistra AJ, de Vries EGE, Mulder NH, Zijlstra JG (1993) Topoisomerase II as a target to VM-26 and 4′-(9-acridinylamino) methanesulfon-m-aniside in atypical multidrug resistant human small cell lung carcinoma cells. Cancer Res 53:1064–1071
Laemmli UK (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lee M-S, Wang JC, Beran M (1992) Two independent amsacrineresistant human myeloid leukemia cell line share an identical point mutation the 170 kDA form of human topoisomerase II. J Mol Biol 223:837–843
Liu LF, Rowe TC, Yang L, Tewey KM, Chen GL (1983) Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem 258:15365–15370
Markman M, George M, Hakes, T, Reichman B, Hoskins W, Rubin S, Jones W, Almandrones L, Lewis JL (1990) Phase II trial of intraperitoneal mitoxantrone in the management of refractory ovarian cancer. J Clin Oncol 8:146–150
Mazzoni A, Trave F, Canti GF, Franco P (1989) Effect of the tiapamil analog RO11-2933 on cellular sensitivity to antitumor drugs in sensitive and multidrug resistant human ovarian cancer cells. Anticancer Res 9:367–372
Mazzoni A, Trave F, Russo P, Nicolin A, Rustum YM (1990) Generation and characterization of a low-degree drug-resistant human tumor cell line. Oncology 47:488–494
Noviello E, Aluigi M, Cimoli G, Rovini E, Mazzoni A, Parodi S, De Sessa F, Russo P (1994) Sister chromatid exchanges, chromosomal aberrations and cytotoxicity produced by topoisomerase II-targeted drugs in sensitive (WT) and resistant (DX3) human ovarian cancer cells: correlations with the formation of DNA double-strand breaks. Mutat Res 311:21–29
O'Dwyer PJ, LaCreta FP, Daugherty JP (1991) Phase I pharmacokinetic study of intraperitoneal etoposide. Cancer Res 51:2041–2046
Perez RP, Hamilton TC, Ozols RF, Young RC (1993) Mechanisms and modulation of resistance to chemotherapy in ovarian cancer. Cancer 71:1571–1577
Pommier Y, Betrand R (1993) The mechanisms of formation of chromosomal abberations: role of eukaryotic DNA topoisomerases. In: Kirsch IR (ed) The causes and consequences of chromosomal aberrations. CRC, Boca Raton, Fla, pp 277–309
Pommier Y, Minford JK, Schwartz RE, Zwelling LA, Kohn KW (1985) Effects of the intercalators 4′-(9-acridinylamino) methanesulfon-m-anisidide and 2-methyl-9-hydroxyellipticinium on topoisomerase II mediated DNA strand cleavage and strand passage. Biochemistry 14:6410–6416
Pommier Y, Kerrigan D, Schwartz RE, Swack JA, McCurdy A (1986) Alterated DNA topoisomerase II activity in Chinese hamster cells resistant to topoisomerase II inhibitors. Cancer Res 46:3075–3081
Potmesil M, Hsiang Y-H, Liu LF, Bank B, Grossberg H, Kirschenbaum S, Forlenzar TJ, Penziner A, Kanganis D, Knowles D, Traganos F, Silber R (1988) Resistance of human leukemic and normal lymphocytes to drug-induced DNA cleavage and low levels of DNA topoisomerase II. Cancer Res 48:3537
Russo P, Poggi L, Parodi S, Pedrini AM, Kohn KW, Pommier Y (1991) Production of protein-associated DNA breaks by 8-methoxycaffeine, caffeine and 8-chlorocaffeine in isolated nuclei from L1210 cells: comparison with those produced by topoisomerase II inhibitors. Carcinogenesis 12:1781–1790
Sinha BK, Haim N, Dusre L, Kerrigan D, Pommier Y (1988) DNA strand breaks produced by etoposide (VP-16,213) in sensitive and resistant human breast tumor cells: implications for the mechanism of action. Cancer Res 48:5096–5100
Utsugi T, Mattern MR, Mirabelli CK, Hanna N (1990) Potentiation of topoisomerase inhibitor-induced DNA strand breakage and cytotoxicity by tumor necrosis factor: enhancement of topoisomerase activity as a mechanism of potentiation. Cancer Res 50:2636–2640
Valenti M, Cimoli G, Mariani GL, Conte PF, Parodi S, Russo P (1993) Potentiation of TNF-mediated cell killing by mitoxantrone: relationship to DNA-single-strand breaks formation. Biochem Pharmacol 46:1199–1206
Van Der Bliek AM, Borst P (1989) Multidrug, resistance. Adv Cancer Res 52:165–203
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Cimoli, G., Valenti, M., Noviello, E. et al. “Atypical” multidrug resistance in human ovarian cancer cell line A2780 selected for resistance to doxorubicin (A2780 DX3). J Cancer Res Clin Oncol 121, 155–163 (1995). https://doi.org/10.1007/BF01198097
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DOI: https://doi.org/10.1007/BF01198097