Potential use of the comet assay in the clinical management of cancer

McKenna, Declan; McKeown, Stephanie; McKelvey‐Martin, Valerie J.

doi:10.1093/mutage/gem054

Cited by 117 publications

(101 citation statements)

References 90 publications

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“…DNA is considered a primary target of ionizing radiation acting both directly or indirectly. The comet assay is often used to quantity DNA damage, which is a marker for radiosensitivity 16) . The majority of DNA damage is repaired within minutes or hours after induction 12,17,18) , but some is not repaired or not repaired properly.…”

Section: Discussionmentioning

confidence: 99%

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

Dobrzyńska¹,

Pachocki²,

Gajowik³

et al. 2014

Journal of Occupational Health

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The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel: Małgorzata M. DOBRZYŃSKA, et al. Department of Radiation Hygiene and Radiobiology, National Institute of Public Health – National Institute of Hygiene, Poland— Objectives The aim of this study was estimation of DNA strand breaks in leukocytes of peripheral blood of staff in a nuclear medicine department. Methods The exposed group consisted of 46 volunteers and the control group consisted of 40 volunteers. Samples consisting of 1 ml whole blood were collected by venepuncture. DNA damage in leukocytes was detected by alkaline comet assay. Results There was no correlation between the effective dose measured by individual dosimeters and DNA damage and no differences between sexes. The mean level of damage to DNA in people exposed to ionizing radiation was significantly elevated compared with control individuals. The highest value for mean comet tail moment was noted in leukocytes of PET/CT and scintigraphy technicians (1.28 vs. 0.30 for control, p=0.013). The levels of DNA damage in leukocytes of workers in category B (effective dose may exceed 1 mSv/year) were significantly enhanced. The DNA migration of leukocytes in exposed smokers and nonsmokers was similar. In the control group the damage to DNA of leukocytes in smokers was markedly but not significantly higher compared with nonsmokers. Conclusions Occupational exposure to ionizing radiation leads to enhanced levels of reversible DNA damage in leukocytes of nuclear medicine employees. The level of DNA damage depends on the kind of work. Cigarette smoking is related to the increase in DNA damage in unexposed individuals but not in nuclear medicine workers. Radiation seems to be a stronger inducer of DNA damage than smoking. Although most of the DNA damage detected by comet assay is repaired, further improvement of radiation safety should be taken under consideration.

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Section: Discussionmentioning

confidence: 99%

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

Dobrzyńska¹,

Pachocki²,

Gajowik³

et al. 2014

Journal of Occupational Health

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“…The present study investigates the effect of oxaliplatin versus satraplatin in terms of interstrand crosslinking action, SCE frequencies in S phase and induction of DNA double-strand breaks detected by γH2AX foci formation in lymphocytes from colorectal cancer patients and healthy individuals in vitro. Peripheral blood lymphocytes appear to be the cells of choice due to the fact that in a variety of cancers, these cells demonstrate basal DNA damage [50] Many chemoand radiotherapeutic agents have consequently been stu0 died to establish increased baseline DNA damage and diminished repair efficiency following exposure in cancer patients [51]. These appear to be the best choice of surrogate human cells due to the fact that the collection of lymphocytes is sufficiently non-invasive in comparison to collection of other target tissues.…”

Section: Discussionmentioning

confidence: 99%

<i>In Vitro</i> Investigation of DNA Damage Induced by the DNA Cross-Linking Agents Oxaliplatin and Satraplatin in Lymphocytes of Colorectal Cancer Patients

Alotaibi¹,

Baumgartner²,

Najafzadeh³

et al. 2012

JCT

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Exposure to toxic chemicals, especially chemotherapeutic drugs, may induce several DNA lesions, including DNA interstrand crosslinks. These crosslinks are considered toxic lesions to the dividing cells since they can induce mutations, chromosomal rearrangements, and cell death. Many DNA interstrand crosslinks lesions can be generated by platinum-based chemotherapeutic agents. Satraplatin is a novel orally administered platinum-based chemotherapeutic agent. In the present study, we investigated DNA interstrand crosslinks lesions induced by oxaliplatin and satraplatin in lymphocytes obtained from colorectal cancer patients and healthy volunteers. Satraplatin demonstrated an increase in interstrand crosslinks in a dose-dependent manner in the Comet assay (p < 0.001). In addition, satraplatin and oxaliplatin increased significantly the number of sister chromatid exchanges up to 8.5-fold and 5.1-fold (p < 0.001) respectively, when treated with 2 µM concentration in comparison to untreated colorectal cancer cells. Further, the γH2AX foci formation was investigated by an immunofluorescence assay with oxaliplatin and satraplatin. The γH2AX foci formation rate was increased by approximately 9-fold when lymphocytes were treated with 2 μM oxaliplatin. Satraplatin was found to significantly induce the number of γH2AX foci by 8.5-fold and 11-fold with both 0.2 μM and 2.0 μM, respectively, compared to the control volunteers that may indicate the repair system in cancer cells experiences a loss of ability to cope with the repair of DSBs. In conclusion, oxaliplatin and satraplatin effectively induced DNA interstrand crosslinks in lymphocytes obtained from colorectal cancer patients and healthy volunteers in vitro. Here, to the best of our knowledge we report for the first time evidence of DNA double strand breaks formation as a possible molecular mechanism of action for satraplatin.

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“…Although most base lesions are not directly detected, base adducts can be detected when converted to abasic sites or single strand breaks with the addition of purified DNA repair enzymes (7)(8)(9)(10)(11). The comet assay has been used in a variety of applications, including genotoxicity testing, human biomonitoring and epidemiology, environmental health, and basic research on DNA damage and repair (7,8,(12)(13)(14)(15)(16)(17)(18)(19). Compared with other DNA damage assays (20)(21)(22), the comet assay is relatively inexpensive, is very sensitive, and can assess the integrated cellular response to many kinds of DNA lesions simultaneously (7,8,19).…”

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confidence: 99%

Single cell trapping and DNA damage analysis using microwell arrays

Wood

Weingeist

Bhatia

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

240

284

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With a direct link to cancer, aging, and heritable diseases as well as a critical role in cancer treatment, the importance of DNA damage is well-established. The intense interest in DNA damage in applications ranging from epidemiology to drug development drives an urgent need for robust, high throughput, and inexpensive tools for objective, quantitative DNA damage analysis. We have developed a simple method for high throughput DNA damage measurements that provides information on multiple lesions and pathways. Our method utilizes single cells captured by gravity into a microwell array with DNA damage revealed morphologically by gel electrophoresis. Spatial encoding enables simultaneous assays of multiple experimental conditions performed in parallel with fully automated analysis. This method also enables novel functionalities, including multiplexed labeling for parallel single cell assays, as well as DNA damage measurement in cell aggregates. We have also developed 24-and 96-well versions, which are applicable to high throughput screening. Using this platform, we have quantified DNA repair capacities of individuals with different genetic backgrounds, and compared the efficacy of potential cancer chemotherapeutics as inhibitors of a critical DNA repair enzyme, human AP endonuclease. This platform enables high throughput assessment of multiple DNA repair pathways and subpathways in parallel, thus enabling new strategies for drug discovery, genotoxicity testing, and environmental health.base excision repair | comet assay | DNA repair D NA damage is a critical risk factor for cancer, aging, and heritable diseases (1-3), and it is the underlying basis for most frontline cancer therapies (4). Increased knowledge about DNA damage and repair would facilitate disease prevention, identification of individuals at increased risk of cancer, discovery of novel therapeutics, and better drug safety testing. Despite their obvious translational impact, most DNA damage assays have not changed significantly in more than two decades, and thus are not compatible with modern high throughput screening technologies. To better assess the biological impact of damage, we need data that reflect the integrated effect of multiple DNA repair pathways and subpathways, in response to a range of DNA lesions, measured in an accurate and high throughput fashion.The single cell gel electrophoresis or "comet" assay is based on the principle that relaxed loops (induced by single strand breaks) and DNA fragments migrate farther in an agarose gel than undamaged DNA (5-8). The alkali comet assay sensitively detects a range of DNA lesions, including strand breaks (single and double), as well as alkali sensitive sites. Although most base lesions are not directly detected, base adducts can be detected when converted to abasic sites or single strand breaks with the addition of purified DNA repair enzymes (7-11). The comet assay has been used in a variety of applications, including genotoxicity testing, human biomonitoring and epidemiology, environmental healt...

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Potential use of the comet assay in the clinical management of cancer

Cited by 117 publications

References 90 publications

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

<i>In Vitro</i> Investigation of DNA Damage Induced by the DNA Cross-Linking Agents Oxaliplatin and Satraplatin in Lymphocytes of Colorectal Cancer Patients

Single cell trapping and DNA damage analysis using microwell arrays

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Potential use of the comet assay in the clinical management of cancer

Cited by 117 publications

References 90 publications

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

The Effect Occupational Exposure to Ionizing Radiation on the DNA Damage in Peripheral Blood Leukocytes of Nuclear Medicine Personnel

&lt;i&gt;In Vitro&lt;/i&gt; Investigation of DNA Damage Induced by the DNA Cross-Linking Agents Oxaliplatin and Satraplatin in Lymphocytes of Colorectal Cancer Patients

Single cell trapping and DNA damage analysis using microwell arrays

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<i>In Vitro</i> Investigation of DNA Damage Induced by the DNA Cross-Linking Agents Oxaliplatin and Satraplatin in Lymphocytes of Colorectal Cancer Patients