Topoisomerase II inhibition and altered kinetics of formation and repair of nitrosourea and cisplatin-induced DNA interstrand cross-links and cytotoxicity in human glioblastoma cells.

By altering the accessibility of DNA sequences for alkylation or platination, and/or for subsequent repair, topoisomerase II can potentially affect the level of DNA interstrand cross-links induced in cells by bifunctional agents. In this study, we investigated the extent to which inhibition of topoisomerase II activity in a human glioblastoma multiforme cell line alters the kinetics of both the formation and the repair of total genomic DNA interstrand cross-links, as well as the sensitivity of the tumor cells to cis-diamminedichloroplatinum II (cis-DDP) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). Cells were incubated with and without 200 microM novobiocin, a known topoisomerase II inhibitor, for 24 h, followed by exposure to 50 microM BCNU and 25 microM cis-DDP. DNA interstrand cross-linking was determined at various time points over 72 h, using a modified ethidium bromide-DNA binding assay. Sensitivity of the cells to cis-DDP and BCNU was also determined with and without novobiocin pretreatment with 200 microM novobiocin. This concentration of novobiocin showed no significant direct cytotoxicity, although it inhibited topoisomerase II activity in tumor cell nuclear extracts by 73%. A significant decrease in the rate of repair of both cis-DDP and BCNU induced DNA interstrand cross-links, with a corresponding decrease in the clonogenic survival of the cells, was observed following novobiocin exposure. Although the peak cross-link indices of novobiocin-treated cells relative to controls were not significantly increased, residual DNA cross-linking in the cells after 72 h was increased by 1.4-fold for BCNU and 3-fold for cells treated with cis-DDP, thus, indicating a greater effect of topoisomerase II on cross-link repair than on cross-link formation. These data suggest that inhibition of topoisomerase II may provide a potentially effective clinical strategy for sensitizing human brain tumors, and possibly other tumors as well, to DNA cross-linking anticancer agents.

[1]  P. Kornblith.  Chemotherapy of primary brain tumors , 2005, Journal of Neuro-Oncology.

[2]  D. Siemann,et al.  Modulation of the cell cycle-dependent cytotoxicity of adriamycin and 4-hydroperoxycyclophosphamide by novobiocin, an inhibitor of mammalian topoisomerase II. , 1992, Cancer research.

[3]  K. Kohn,et al.  A fundamental role for cell cycle regulation in the chemosensitivity of cancer cells? , 1992, Seminars in cancer biology.

[4]  B. Teicher,et al.  A phase I clinical trial of novobiocin, a modulator of alkylating agent cytotoxicity. , 1991, Cancer research.

[5]  U. K. Laemmli,et al.  Chromosome assembly in vitro: Topoisomerase II is required for condensation , 1991, Cell.

[6]  M. Barcellos-Hoff,et al.  Differential drug sensitivity conferred by growth status detected in a mixed population of cycling and noncycling cells. , 1990, Cancer research.

[7]  C. Holm,et al.  Segregation of recombined chromosomes in meiosis I requires DNA topoisomerase II , 1990, Cell.

[8]  A. Collins,et al.  Comparison of effects of fostriecin, novobiocin, and camptothecin, inhibitors of DNA topoisomerases, on DNA replication and repair in human cells. , 1990, Nucleic acids research.

[9]  U. K. Laemmli,et al.  Preferential, cooperative binding of DNA topoisomerase II to scaffold‐associated regions. , 1989, The EMBO journal.

[10]  R. Rowley,et al.  Novobiocin, nalidixic acid, etoposide, and 4'-(9-acridinylamino)methanesulfon-m-anisidide effects on G2 and mitotic Chinese hamster ovary cell progression. , 1989, Cancer research.

[11]  L. Liu,et al.  DNA topoisomerase poisons as antitumor drugs. , 1989, Annual review of biochemistry.

[12]  L. Liu,et al.  Topoisomerases: novel therapeutic targets in cancer chemotherapy. , 1988, Biochemical pharmacology.

[13]  S. Jacob,et al.  Inhibition of RNA polymerase I-directed transcription by novobiocin. Potential use of novobiocin as a general inhibitor of eukaryotic transcription initiation. , 1988, The Journal of biological chemistry.

[14]  F. Ali-Osman,et al.  Optimization and characterization of the capillary human tumor clonogenic cell assay. , 1988, Cancer research.

[15]  S. Jacob,et al.  Novobiocin inhibits initiation of RNA polymerase II-directed transcription of the mouse metallothionein-I gene independent of its effect on DNA topoisomerase II. , 1987, Nucleic acids research.

[16]  T. Uemura,et al.  DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe , 1987, Cell.

[17]  W. Ross,et al.  Topoisomerase-specific drug sensitivity in relation to cell cycle progression , 1987, Molecular and cellular biology.

[18]  R. C. Adlakha,et al.  Cell cycle stage dependent variations in drug-induced topoisomerase II mediated DNA cleavage and cytotoxicity. , 1987, Biochemistry.

[19]  S. Dresler,et al.  Direct inhibition of u.v.-induced DNA excision repair in human cells by novobiocin, coumermycin and nalidixic acid. , 1987, Carcinogenesis.

[20]  M. Berger,et al.  PRIMARY CULTURES OF HUMAN GLIAL TUMOR CELLS AND THEIR RELATIONSHIP TO HISTOPATHOLOGY: 119 , 1987 .

[21]  B. Teicher,et al.  Novobiocin enhances alkylating agent cytotoxicity and DNA interstrand crosslinks in a murine model. , 1987, The Journal of clinical investigation.

[22]  R. Sternglanz,et al.  Need for DNA topoisomerase activity as a swivel for DNA replication and for transcription of ribosomal RNA , 1987, Nature.

[23]  B. Sahai,et al.  A quantitative decatenation assay for type II topoisomerases. , 1986, Analytical biochemistry.

[24]  A. Collins,et al.  Novobiocin inhibition of DNA excision repair may occur through effects on mitochondrial structure and ATP metabolism, not on repair topoisomerases. , 1985, Carcinogenesis.

[25]  F. Ali-Osman,et al.  Chemical structure of carbamoylating groups and their relationship to bone marrow toxicity and antiglioma activity of bifunctionally alkylating and carbamoylating nitrosoureas. , 1985, Cancer research.

[26]  H. Vosberg.  DNA topoisomerases: enzymes that control DNA conformation. , 1985, Current topics in microbiology and immunology.

[27]  J. Cleaver,et al.  Novobiocin-mediated inhibition of polymerization and ligation of DNA in vitro. , 1984, Mutation research.

[28]  J. David,et al.  Quinolone antibiotics inhibit eucaryotic DNA polymerase α and β, Terminal deoxynucleotidyl Transferase but not DNA ligase , 1984 .

[29]  T. Brent.  Suppression of cross-link formation in chloroethylnitrosourea-treated DNA by an activity in extracts of human leukemic lymphoblasts. , 1984, Cancer research.

[30]  T. Watanabe,et al.  Inhibition of retrovirus RNA-dependent DNA polymerase by novobiocin and nalidixic acid. , 1983, The Journal of general virology.

[31]  R. Snyder,et al.  Studies on the inhibition of repair of ultraviolet- and methyl methanesulfonate-induced damage in the DNA of human fibroblasts by novobiocin. , 1982, Nucleic acids research.

[32]  G. P. Studzinski,et al.  DNA dependence and inhibition by novobiocin and coumermycin of the nucleolar adenosine triphosphatase (ATPase) of human fibroblasts. , 1982, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[33]  S. Squires,et al.  Inhibitors of repair DNA synthesis. , 1982, Nucleic acids research.

[34]  J. Cleaver.  Specificity and completeness of inhibition of DNA repair by novobiocin and aphidicolin. , 1982, Carcinogenesis.

[35]  H. Wright,et al.  Nalidixic acid, oxolinic acid, and novobiocin inhibit yeast glycyl- and leucyl-transfer RNA synthetases. , 1981, Science.

[36]  D. Scudiero,et al.  Dependence of mammalian DNA synthesis on DNA supercoiling. III. Characterization of the inhibition of replicative and repair-type DNA synthesis by novobiocin and nalidixic acid. , 1981, Biochimica et biophysica acta.

[37]  H. Edenberg.  Novobiocin inhibition of simian virus 40 DNA replication , 1980, Nature.

[38]  M. Colvin,et al.  A chemical basis for the antitumor activity of chloroethylnitrosoureas. , 1976, Biochemical pharmacology.

[39]  R. Tobey.  Different drugs arrest cells at a number of distinct stages in G2 , 1975, Nature.

[40]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.