Formation and persistence of DNA adducts of anticancer drug ellipticine in rats

Stiborová, Marie; Rupertová, Martina; Aimová, Dagmar; Ryšlavá, Helena; Frei, Eva

doi:10.1016/j.tox.2007.03.026

Cited by 30 publications

(62 citation statements)

References 34 publications

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“…1) [21,23]. CYPand/or peroxidase-mediated ellipticine-DNA adducts were detected also in rats and mice in vivo [18,22,24]. The same DNA adducts were also detected in cells in culture expressing enzymes activating ellipticine (CYP1A1, COX-1 and MPO), such as human breast adenocarcinoma MCF-7 cells [25], leukemia HL-60 and CCRF-CEM cells [26] and V79…”

Section: ) An Alkaloid Isolated Frommentioning

confidence: 87%

“…5D), in several organs of rats [17,22] and mice [24] exposed to this agent. Both these adducts were found to be generated from 13-hydroxy-and 12-hydroxyellipticine [19,21,41] (Fig.…”

Section: Determination Of Dna Adduct Formation By Ellipticine In Neurmentioning

confidence: 97%

“…We have demonstrated that ellipticine also covalently binds to DNA in vitro and in vivo after being enzymatically activated with cytochromes P450 (CYP) or peroxidases [3,[17][18][19][20][21][22][23],…”

Section: ) An Alkaloid Isolated Frommentioning

confidence: 99%

“…The three parental neuroblastoma cell lines shown to be sensitive to ellipticine (Table 1) were treated with 0.1, 1, and 10 M ellipticine for 48 h. Using the nuclease P1 version of 32 Ppostlabeling assay, which was found to be suitable to detect and quantify DNA adducts formed by ellipticine [3,[17][18][19]21,22], ellipticine-derived adducts were detected in the DNA of these cells (Fig. 5A-C).…”

Section: Determination Of Dna Adduct Formation By Ellipticine In Neurmentioning

confidence: 99%

“…Because CYPs and peroxidases, which activate ellipticine [3,17,19,22], were found to be expressed in neuroblastoma cells (see above), we evaluated whether S9 fractions isolated from the parental IMR-32, UKF-NB-3 and UKF-NB-4 cells activate ellipticine to species forming DNA adducts.…”

Section: S9 Fractions Of Neuroblastoma Cells Are Capable Of Activatinmentioning

confidence: 99%

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The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells

Poljaková

Eckschlager

Hraběta

et al. 2009

Biochemical Pharmacology

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Ellipticine is an antineoplastic agent, whose mode of action is based mainly on DNA intercalation, inhibition of topoisomerase II and formation of covalent DNA adducts mediated by cytochromes P450 and peroxidases. Here, the molecular mechanism of DNA-mediated ellipticine action in human neuroblastoma IMR-32, UKF-NB-3 and UKF-NB-4 cancer cell lines was investigated. Treatment of neuroblastoma cells with ellipticine resulted in apoptosis induction, which was verified by the appearance of DNA fragmentation, and in inhibition of cell growth. These effects were associated with formation of two covalent ellipticine-derived DNA adducts, identical to those formed by the cytochrome P450-and peroxidase-mediated ellipticine metabolites, 13-hydroxy-and 12-hydroxyellipticine. The expression of these enzymes at mRNA and protein levels and their ability to generate ellipticine-DNA adducts in neuroblastoma cells were proven, using the real-time polymerase chain reaction, Western blotting analyses and by analyzing ellipticine-DNA adducts in incubations of this drug with neuroblastoma S9 fractions, enzyme cofactors and DNA. The levels of DNA adducts correlated with toxicity of ellipticine to IMR-32 and UKF-NB-4 cells, but not with that to UKF-NB-3 cells. In addition, hypoxic cell culture conditions resulted in a decrease in ellipticine toxicity to IMR-32 and UKF-NB-4 cells and this correlated with lower levels of DNA adducts. Both these cell lines accumulated in S phase, suggesting that ellipticine-DNA adducts interfere with DNA replication. The results demonstrate that among the multiple modes of ellipticine antitumor action, formation of covalent DNA adducts by ellipticine is the predominant mechanism of cytotoxicity to IMR-32 and UKF-NB-4 neuroblastoma cells.

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Section: ) An Alkaloid Isolated Frommentioning

confidence: 87%

“…5D), in several organs of rats [17,22] and mice [24] exposed to this agent. Both these adducts were found to be generated from 13-hydroxy-and 12-hydroxyellipticine [19,21,41] (Fig.…”

Section: Determination Of Dna Adduct Formation By Ellipticine In Neurmentioning

confidence: 97%

Section: ) An Alkaloid Isolated Frommentioning

confidence: 99%

Section: Determination Of Dna Adduct Formation By Ellipticine In Neurmentioning

confidence: 99%

Section: S9 Fractions Of Neuroblastoma Cells Are Capable Of Activatinmentioning

confidence: 99%

See 3 more Smart Citations

The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells

Poljaková

Eckschlager

Hraběta

et al. 2009

Biochemical Pharmacology

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Study of DNA–ellipticine interaction by capillary electrophoresis with laser‐induced fluorescence detection

et al. 2012

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Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazole), an alkaloid isolated from Apocynaceae plants, exhibits an antitumor activity, which is exceptionally high against several specific types of tumors. Ellipticine is also interesting as an anticancer drug as it has limited side effects and lacks of hematological toxicity. Various methods to study intercalating activity of this drug have been developed. However, to our best knowledge, capillary electrophoresis (CE) as a technique combining high separation resolution with various detection options has never been used for these purposes. In this study, a novel separation method based on CE with laser-induced fluorescence (CE-LIF) detection has been developed for the determination of ellipticine and for the monitoring of ellipticine-DNA interaction. Sodium acetate (50 mM, pH 4.5) was used as a background electrolyte and LIF detection at λ(ex) = 488 nm. The limit of detection for ellipticine was determined to be 5 × 10⁻⁸ M. A total of 20% dimethyl sulfoxide was found optimal as sample solvent. Additionally, intercalation of ellipticine into the double-stranded DNA was investigated. Signal corresponding to ellipticine was decreasing and a new peak appeared and was growing. It can be concluded that CE-LIF is a method applicable to in vitro studies of ellipticine-DNA complexes.

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Cytochrome P450 system expression and DNA adduct formation in the liver of Zacco platypus following waterborne Benzo(a)pyrene exposure: implications for biomarker determination

Lee

Kim

Yoon

et al. 2012

Environmental Toxicology

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Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon that causes mutations and tumor formation. Zacco platypus is a sentinel species that is suitable for monitoring aquatic environments. We studied cytochrome P450 system (CYP system) expression and DNA adduct formation in the liver of Z. platypus following waterborne exposure to BaP. The results showed both dose and time dependency. The significant induction levels of CYP system mRNA and protein reached maximums at 2 days and 14 days, respectively, and hepatosomatic index was maximally induced at 4 days during 14 days BaP exposure. DNA adduct formation was significantly induced compared to corresponding controls (t-test, p < 0.01) after 4 days of exposure in 100 μg/L BaP. These results indicate that the only use of mRNA expression level of CYP system as a biomarker make us underestimate prolonged toxicity (4-14 days) of BaP and the only use of protein expression level of CYP system make us underestimate acute toxicity (1-2 days) of BaP. Therefore, we suggests that a combinational use of the mRNA expression level and protein expression level of CYP system, hepatosomatic index is a useful biomarker in risk assessment of waterborne BaP exposure. In addition, DNA adduct formation was a useful biomarker in risk assessment of waterborne BaP exposure at 4 days. CYP1A was a more sensitive biomarker than CYP reductase for BaP exposure when considering both the mRNA and protein level. Furthermore, our results show that Z. platypus is a useful species for assessing the risk of waterborne BaP exposure.

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Formation and persistence of DNA adducts of anticancer drug ellipticine in rats

Cited by 30 publications

References 34 publications

The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells

The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells

Study of DNA–ellipticine interaction by capillary electrophoresis with laser‐induced fluorescence detection

Cytochrome P450 system expression and DNA adduct formation in the liver of Zacco platypus following waterborne Benzo(a)pyrene exposure: implications for biomarker determination

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