4-quinolone antibiotics: Positive genotoxic screening tests despite an apparent lack of mutation induction

Bredberg, Anders; Brant, Marta; Riesbeck, Kristian; Azou, Yannick; Forsgren, Arne

doi:10.1016/0027-5107(89)90117-6

Cited by 49 publications

(29 citation statements)

References 20 publications

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“…Studies are in progress to elucidate the mechanism(s) of CPFX-mediated attenuation of γH2AX. Our findings differ from two early studies by Bredberg et al, 16,37 who reported a significant induction of DSBs in Raji lymphoblastoid cells treated for 2 h with 80 mg/ml CPFX. These authors used a limited DNA elution technique, whereas, we used histone γH2AX, a molecular marker that is closely associated with the induction of DSBs.…”

Section: Discussioncontrasting

confidence: 99%

Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not dependent on DNA double-strand break formation

Smart

Halicka²,

Traganos³

et al. 2008

Cancer Biology & Therapy

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Drugs developed for the treatment of conditions other than neoplasia can also show promise as potential antitumor agents. The fluoroquinolone antibiotic ciprofloxacin (CPFX) is known to modulate cycle cell progression and apoptosis in cancer cells, and is thought to induce DNA double-strand breaks (DSBs) via topoisomerase II (topo II) inhibition and stabilized cleavage complex (SCC) formation. DSBs trigger Ser-139 phosphorylation of histone H2AX (γH2AX) by PI-3-like kinases including ATM; γH2AX can serve as a marker of DNA damage when measured in situ using immunocytochemistry and flow cytometry. The aim of the present study was to investigate the relationship between CPFX-mediated DNA damage and induction of apoptosis in human lymphoblastoid cells and phytohaemagglutinin (PHA)-stimulated lymphocytes (Lymphs). Treatment of TK6 cells (wild-type p53) with 100 mg/ml CPFX for 2-10 h produced no increase in γH2AX; to the contrary, its level in S phase cells was reduced at 10 h compared to controls. Nevertheless, stabilization of topo IIα, ATM Ser-1981 phosphorylation and G 2 arrest was observed in TK6 cells exposed to CPFX for ≥4 h. However, following 24 h treatment, γH2AX was dramatically increased in a sub-population of cells indicating the onset of apoptosis (confirmed by presence of activated caspase 3). CPFX had a similar lack of effect on induction of γH2AX at early time points in WTK1 and NH32 cells (devoid of functional p53) and proliferating Lymphs, however, induction of apoptosis was less pronounced than in TK6 cells. Formation of SCC and activation of ATM (but lack of γH2AX induction) indicates topo II-mediated chromatin or DNA changes in the absence of DSBs; ATM activation apparently triggers the G 2 M checkpoint leading to G 2 arrest. The subsequent induction of apoptosis appears to be facilitated by functional p53. CPFX may therefore have a potential use as a chemotherapeutic agent in the treatment of lymphoblast-derived cancer.

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Section: Discussioncontrasting

confidence: 99%

Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not dependent on DNA double-strand break formation

Smart

Halicka²,

Traganos³

et al. 2008

Cancer Biology & Therapy

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“…However, the literature on this subject has presented controversial results (e.g., see references 3, 6, 7, and 11), with some in vitro tests giving an indication of genotoxic damage formation and others providing no such evidence. We have previously reported that human lymphoblastoid cell DNA is broken by ciprofloxacin in culture (3), illustrating that this drug has the capacity to affect cellular DNA in vitro. The results of the present study indicate that the major part of ciprofloxacin-induced DNA breakage can also be seen under nondenaturing conditions.…”

Section: Discussionmentioning

confidence: 92%

Ciprofloxacin-induced inhibition of topoisomerase II in human lymphoblastoid cells

Bredberg

Brant

Jaszyk

1991

Antimicrob Agents Chemother

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The antibacterial activities of the fluorinated 4-quinolones (e.g., ciprofloxacin) have been ascribed to a marked inhibition of bacterial DNA gyrase. In contrast, the influence on purified mammalian DNA enzymes, including topoisomerases, has been reported to be several orders of magnitude weaker, occurring at concentrations higher than 100 ,ug of ciprofloxacin per ml. In this study, using a nondenaturing filter elution method, a marked induction of double-strand DNA breaks in human lymphoblastoid cells exposed to 80 ,ug of ciprofloxacin per ml was seen. The proportion of single-strand versus double-strand DNA breaks was similar to that seen with the topoisomerase II inhibitory antitumor agent VP-16. The cellular recovery was more rapid after treatment with ciprofloxacin than after treatment with VP-16, displaying a normal elution profile within 15 min at 37°C (60 min for VP-16). These data indicate that ciprofloxacin has an effect on intracellularly located topoisomerase H in humans.The antibacterial activities of fluorinated 4-quinolones have been ascribed to the binding of these drugs to DNA (12), leading to a marked inhibition of bacterial DNA gyrase (6). In contrast, the influence on mammalian DNA topoisomerases is several orders of magnitude weaker, as measured by a purified eukaryotic topoisomerase II (topo II) plus plasmid DNA assay (5,8). In those studies an inhibitory influence on the catalytic function of topo II by ciprofloxacin was noted at minimally effective concentrations of 140 ,ug/ml (5) or 150 ,ug/ml (8). Also, topo II-dependent DNA breakage has been assayed by using a plasmid system; cleavage of nonradioactively labeled plasmid DNA was not seen at ciprofloxacin levels up to 1,000 ,ug/ml, whereas in a more sensitive assay with radioactively labeled plasmid, cleavage was observed at 120 ,ug/ml (5).In the present study we investigated the effects of ciprofloxacin on intracellularly located topo II using a human lymphoblastoid cell system. The previously reported (3) cellular DNA breakage associated with ciprofloxacin was determined to be due mainly to the introduction of doublestrand DNA breaks (DSBs), indicating a causative role of topo 11 (13). The proportion of single-strand DNA breaks (SSBs) versus DSBs was similar to that seen with the epipodophyllotoxin antitumor agent 14). However, the cellular recovery from the DNA damage was more rapid with ciprofloxacin than with VP-16, displaying a normal elution profile within 15 min at 37°C (60 min for VP-16).MATERIALS AND METHODS Drugs. Fresh solutions of preservative-free ciprofloxacin (Bayer, Wuppertal, Federal Republic of Germany) were used. Etoposide (VP-16) was obtained from Bristol-Myers (Syracuse, N.Y.) as a concentrated solution for infusion (20 mg/ml).UV irradiation. A total of 106 cells suspended in 4 ml of phosphate-buffered saline (pH 7.4) in a 5-cm-diameter plastic dish were exposed (dish lid off) to radiation from an ordinary laboratory hood UV tube (GTE Sylvania G30 T8) emitting 1.0 W of UVC per m2 at a distance of about 30 cm (as * ...

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“…was confirmed in a separate study in which the compound, when tested at 5 ,ug/ml, induced a positive UDS response in human lymphocytes (6). A similar evaluation of norfloxacin and fleroxacin (AM-833) showed that they were not mutagenic, did not induce DNA strand breakage, and did not elicit a UDS response in either human or mouse skin fibroblasts (25).…”

Section: Reported Effects Of Quinolones On Eucaryotic Topoisomerases mentioning

confidence: 83%

“…Ciprofloxacin at 20 jig/ml was shown to inhibit the progression of mitogen-stimulated lymphocytes through the S and G2/M stages of the cell cycle and to decrease the secretion of immunoglobulins G and M in pokeweed mitogen-stimulated B cells (21). The (6,20,21 (30) and the unscheduled DNA synthesis (UDS) test, the latter of which is run with quinolones incubated in rat hepatocyte cell cultures and also with rat hepatocytes removed from animals dosed with the test quinolone and analyzed directly (38). A number of quinolones are inhibitory in some of these in vitro genotoxicity tests, although positive in vivo tests are rare (Table 1).…”

Section: Reported Effects Of Quinolones On Eucaryotic Topoisomerases mentioning

confidence: 99%

“…As mentioned above, the alkaline elution test measures DNA strand breakage (30) and has been used to test quinolones for this activity, as described by Bredberg et al (6). The cellular DNA is prelabeled with [3H]thymidine and, after incubation with a quinolone for 24 h, the cells are washed and lysed on a 2-,um-pore-size filter.…”

Section: Reported Effects Of Quinolones On Eucaryotic Topoisomerases mentioning

confidence: 99%

See 1 more Smart Citation

Inhibitory effects of quinolone antibacterial agents on eucaryotic topoisomerases and related test systems

Gootz

Barrett

Sutcliffe

1990

Antimicrob Agents Chemother

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4-quinolone antibiotics: Positive genotoxic screening tests despite an apparent lack of mutation induction

Cited by 49 publications

References 20 publications

Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not dependent on DNA double-strand break formation

Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not dependent on DNA double-strand break formation

Ciprofloxacin-induced inhibition of topoisomerase II in human lymphoblastoid cells

Inhibitory effects of quinolone antibacterial agents on eucaryotic topoisomerases and related test systems

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