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

Poljaková, Jitka; Eckschlager, Tomáš; Hraběta, Jan; Hřebačková, Jana; Smutný, S; Frei, Eva; Martínek, Václav; Kizek, René; Stiborová, Marie

doi:10.1016/j.bcp.2009.01.021

Cited by 62 publications

(90 citation statements)

References 47 publications

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“…We found that exposure of human neuroblastoma IMR-32, UKF-NB-3 and UKF-NB-4 cell lines to this agent resulted in strong inhibition of cell growth, followed by induction of apoptosis 7 . These effects were associated with formation of two covalent ellipticine-derived DNA adducts, identical to those formed by the cytochrome P450 (CYP)-and peroxidase-mediated ellipticine metabolites, 13-hydroxy-and 12-hydroxyellipticine (Fig.…”

Section: Introductionmentioning

confidence: 87%

“…DNA from cells treated with ellipticine and doxorubicin was isolated by the phenol-chloroform extraction as described 7 .…”

Section: Dna Isolationmentioning

confidence: 99%

“…2B). Even though ellipticine and doxorubicin are cytotoxic to neuroblastomas 7 , the question arises whether the covalent DNA adduct formation is the predominant mechanism responsible for this cytotoxic effect. To answer this question, development of methods suitable for detecting and quantifying covalent DNA adducts formed by ellipticine and doxorubicin is the first and key target of the research.…”

Section: Introductionmentioning

confidence: 99%

“…P-postlabeling of ellipticine-derived DNA adducts The 32 P-postlabeling of nucleotides using nuclease P1 enrichment procedure 31 , found previously to be appropriate to detect and quantify ellipticine-derived DNA adducts formed in vitro and in vivo 12,14,33,34 was used previously to detect and quantify the adducts formed in neuroblastoma cells 7 . DNA isolated from neuroblastoma cells as described in that study 7 and DNA from experiments performed earlier, namely, calf thymus DNA incubated with 13-hydroxy-and 12-hydroxyellipticine 33,34 were labeled with 32 P to show and compare adduct spot patterns.…”

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confidence: 99%

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Analysis of covalent ellipticine- and doxorubicin-derived adducts in DNA of neuroblastoma cells by the <sup>32</sup>P-postlabeling technique

Stiborová¹,

Poljaková²,

Eckschlager³

et al. 2012

BIOMED PAP

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eBackground. Ellipticine and doxorubicin are antineoplastic agents, whose action is based mainly on DNA damage such as intercalation, inhibition of topoisomerase II and formation of covalent DNA adducts. The key target to resolve which of these mechanisms are responsible for ellipticine and doxorubicin anticancer effects is the development of suitable methods for identifying their individual DNA-damaging effects. Here, the 32 P-postlabeling method was tested to detect covalent DNA adducts formed by ellipticine and doxorubicin. Methods. The standard procedure of 32 P-postlabeling assay, this procedure under ATP-deficient conditions, the version using extraction of adducts with n-butanol and the nuclease P1 enrichment version were used to analyze ellipticineand/or doxorubicin-derived DNA adducts. Results. Two covalent ellipticine-derived DNA adducts, which are associated with cytotoxicity of ellipticine to human UKF-NB-3 and UKF-NB-4 neuroblastoma cell lines, were detected by the 32 P-postlabeling method. These adducts are identical to those formed by the ellipticine metabolites, 13-hydroxy-and 12-hydroxyellipticine. In contrast, no covalent adducts formed by doxorubicin in DNA of these neuroblastoma cells and in DNA incubated with this drug and formaldehyde in vitro were detectable by the 32 P-postlabeling assay. Conclusions. The results presented in this paper are the first to demonstrate that in contrast to covalent DNA adducts formed by ellipticine, the adducts generated by formaldehyde-mediated covalent binding of doxorubicin to DNA are not detectable by the 32 P-postlabeling assay. No DNA adducts were, detectable either in vitro, in incubations of DNA with doxorubicin or in DNA of neuroblastoma cells treated with this drug. The results also suggest that covalent binding of ellipticine to DNA of UKF-NB-3 and UKF-NB-4 neuroblastoma cell lines is the predominant mechanism responsible for the cytotoxicity of this drug. To understand the mechanisms of doxorubicin anticancer effects on neuroblastoma cells, development of novel methods for identifying covalent doxorubicin-derived DNA adducts is the major challenge for further research.

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Section: Introductionmentioning

confidence: 87%

“…DNA from cells treated with ellipticine and doxorubicin was isolated by the phenol-chloroform extraction as described 7 .…”

Section: Dna Isolationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Analysis of covalent ellipticine- and doxorubicin-derived adducts in DNA of neuroblastoma cells by the <sup>32</sup>P-postlabeling technique

Stiborová¹,

Poljaková²,

Eckschlager³

et al. 2012

BIOMED PAP

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“…The sensitivity of cell lines to CDDP was determined by using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test (22). The IC 50 was assessed using an MTT dye reduction assay as described in a previous report (23). The IC 50 values were calculated from at least 3 independent experiments using linear regression of the dose-log response curves using SoftMax Pro microplate data software (Molecular Devices, Sunnyvale, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Changes in MYCN expression in human neuroblastoma cell lines following cisplatin treatment may not be related to MYCN copy numbers

et al. 2013

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Abstract. Neuroblastoma is a tumor accounting for approximately 10% of all childhood malignancies and 50% of all childhood cancer-related deaths. MYCN gene copy number variation represents the most important prognostic factor in neuroblastoma. Prognostic significance of MYCN gene expression is more complicated and may depend on other factors such as MYCN gene copy number status. In the present study, we assessed MYCN gene expression using real-time RT-PCR following cisplatin treatment in three human neuroblastoma cell lines (UKF-NB-3, UKF-NB-4 and SK-N-AS) and their cisplatin-resistant counterparts. We also examined MYCN gene status and copy number (gain and amplification) variations using interphase and metaphase fluorescent in situ hybridization (FISH) and multiplex ligation-dependent probe amplification (MLPA). Only cisplatin-sensitive UKF-NB-4 cells exhibited decreased MYCN expression following treatment with cisplatin. Other sensitive neuroblastoma cells did not exhibit a change in MYCN expression. In contrast, cisplatin-resistant UKF-NB-4 and SK-N-AS cells exhibited increased MYCN expression irrespective of the number of MYCN copies or concentration of cisplatin in the medium. In MYCN-amplified neuroblastoma cells we did not observe any significant change in the number of MYCN copies after cisplatin treatment, whereas MYCN-non-amplified SK-N-AS cells revealed during cisplatin treatment an increased number of MYCN gene copies caused by 2p gain in the majority of cells by FISH. We postulated that cisplatin treatment does not result directly in altered transcription of MYCN. A functional change in MYCN mRNA levels and increased MYCN expression in cisplatin-resistant neuroblastoma cells do not have a clear relationship to MYCN copy numbers. These findings may further contribute to the understanding of cisplatin chemotherapy in connection with MYCN expression, and the possible copy number variations in MYCN neuroblastoma cells may be of importance since targeting of MYCN is being tested as neuroblastoma therapy.

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Bioactivities and Action Mechanisms of Ellipticine Derivatives Reported Prior to 2023

Hou,

Xu,

Huang

et al. 2024

Chemistry & Biodiversity

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Currently, natural products are one of the priceless options for finding novel chemical pharmaceutical entities. Ellipticine is a naturally occurring alkaloid isolated from the leaves of Ochrosia elliptica Labill. Ellipticine and its derivatives are characterized by multiple biological activities. The purpose of this review was to provide a critical and systematic assessment of ellipticine and its derivatives as bioactive molecules over the last 60 years. Publications focused mainly on the total synthesis of alkaloids of this type without any evaluation of bioactivityhave been excluded. We have reviewed papers dealing with the synthesis, bioactivity evaluation and mechanism of action of ellipticine and its derivatives. It was found that ellipticine and its derivatives showed cytotoxicity, antimicrobial ability, and anti‐inflammatory activity, among which cytotoxicity toward cancer cell lines was the most investigated aspect. The inhibition of DNA topoisomerase II was the most relevant mechanism for cytotoxicity. The PI3K/AKT pathway, p53 pathway, and MAPK pathway were also closely related to the antiproliferative ability of these compounds. In addition, the structure‐activity relationship was deduced, and future prospects were outlined. We are confident that these findings will lay a scientific foundation for ellipticine‐based drug development, especially for anticancer agents.

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

Cited by 62 publications

References 47 publications

Analysis of covalent ellipticine- and doxorubicin-derived adducts in DNA of neuroblastoma cells by the <sup>32</sup>P-postlabeling technique

Analysis of covalent ellipticine- and doxorubicin-derived adducts in DNA of neuroblastoma cells by the <sup>32</sup>P-postlabeling technique

Changes in MYCN expression in human neuroblastoma cell lines following cisplatin treatment may not be related to MYCN copy numbers

Bioactivities and Action Mechanisms of Ellipticine Derivatives Reported Prior to 2023

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