Assessment of Radiation Induced DNA Damage in Human Peripheral Blood Lymphocytes Using COMET Assay

Kaur, Satbir; Sangeeta, Sangeeta; Galhna, Kiranjeet Kaur; Gautam, Nisha

doi:10.21276/ijlssr.2017.3.4.17

Cited by 7 publications

(5 citation statements)

References 24 publications

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“…The differences between the exposure types were also evident, showing statistical differences, specifically higher sensitivity to DNA damage in BRCA1 VUS carriers. The effect observed for irradiated samples demonstrates that this assay is a viable method for evaluating primary DNA lesions ( 49 ), suggesting a potential role of this genetic variant in hampering the repair mechanisms.…”

Section: Discussionmentioning

confidence: 94%

BRCA1 VUS: A functional analysis to differentiate pathogenic from benign variants identified in clinical diagnostic panels for breast cancer

et al. 2023

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Genetic testing for susceptibility genes through next-generation sequencing (NGS) has become a widely used technique. Using this, a number of genetic variants have been identified, several of which are variants of unknown significance (VUS). These VUS can either be pathogenic or benign. However, since their biological effect remains unclear, functional assays are required to classify their functional nature. As the use of NGS becomes more mainstream as a diagnostic tool in clinical practice, the number of VUS is expected to increase. This necessitates their biological and functional classification. In the present study, a VUS was identified in the BRCA1 gene (NM_007294.3:c.1067A>G) in two women at risk for breast cancer, for which no functional data has been reported. Therefore, peripheral lymphocytes were isolated from the two women and also from two women without the VUS. DNA from all samples were sequenced by NGS of a breast cancer clinical panel. Since the BRCA1 gene is involved in DNA repair and apoptosis, the functional assays chromosomal aberrations, cytokinesis-blocked micronucleus, comet, γH2AX, caspase and TUNEL assays were then conducted on these lymphocytes after a genotoxic challenge by ionizing radiation or doxorubicin to assess the functional role of this VUS. The micronucleus and TUNEL assays revealed a lower degree of DNA induced-damage in the VUS group compared with those without the VUS. The other assays showed no significant differences between the groups. These results suggested that this BRCA1 VUS is likely benign, since the VUS carriers were apparently protected from deleterious chromosomal rearrangements, subsequent genomic instability and activation of apoptosis.

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Section: Discussionmentioning

confidence: 94%

BRCA1 VUS: A functional analysis to differentiate pathogenic from benign variants identified in clinical diagnostic panels for breast cancer

et al. 2023

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“…It is well known that ionizing radiation can directly cause oxidative stress by accumulating energy in cells or increase the formation of free radicals, and oxidative stress can indirectly induce DNA damage [16].…”

Section: Discussionmentioning

confidence: 99%

Bilgisayarlı Tomografi İle Yapılan Perkütan Torasik Kitle Biyopsilerinde Lenfositlerdeki DNA Hasarının Değerlendirilmesi

Ayyildiz

ASLANKOÇ

Saygın

et al. 2023

Süleyman Demirel Üniversitesi Sağlık Bilimleri Dergisi

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Purpose: The increase in the medical and industrial uses of radiation in the last century has caused people to be exposed to higher doses of radiation. Although the harms of high-dose radiation on human health are known, the effects of low-dose radiation on health have not yet been fully elucidated. In this study, we aimed to investigate DNA damage in lymphocytes in patients with lung, pleura/thoracic wall masses planned for percutaneous thoracic mass biopsy with computed tomography (CT). Methods: Sixteen patients referred to the Radiology Clinic of a public institution hospital with lung, pleura/thoracic wall masses and scheduled to undergo a CT-guided percutaneous biopsy were included in the study. All the biopsies were performed with a 128-slice CT device (Definition AS, Siemens Medical Solutions, Forchheim, Germany). Lymphocytes were analyzed using the comet assay in the venous blood samples taken from the patients before and after the biopsy procedure. DNA damage was quantitatively evaluated with the imaging analysis method. Results: In the CT analysis data of the study group, the mean scan distance was found to be 19.92±3.60 sec, the mean total milliampere-seconds was 807.43±304.51, and the mean dose-length product was 765.44±278.36 mGy.cm. The mean comet score was 200.50±40.54 for the cells that migrated before the procedure and 237.37±27.85 for those migrating after the procedure. The post-procedure comet scores significantly increased compared to the pre-procedure comet scores (p=0.038). Conclusion: Post-procedure DNA damage was detected in patients who underwent CT-guided percutaneous biopsy.

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“…Ionizing radiation has become an essential part of medicine, both for diagnostic and therapeutic purposes [1,2]. Medical radiation workers, such as radiotherapy operators, risk exposure to ionizing radiation despite using radiation protection devices.…”

Section: Introductionmentioning

confidence: 99%

“…Peripheral blood lymphocytes have a high sensitivity to ionizing radiation, so they are often used as a parameter for DNA damage. One of the fast and sensitive visual methods to quantitatively detect DNA damage in cells is the comet assay [2,11]. The alkaline comet assay can detect DNA damage up to a minimum dose of 0.6 cGy [12].…”

Section: Introductionmentioning

confidence: 99%

Adaptive Response of Peripheral Blood Lymphocytes in Medical Radiation Workers using the Comet Assay: A Preliminary Studyomet Assay: A Preliminary Study

Kisnanto

Darlina

Musthafa³

et al. 2022

Indonesian Journal of Cancer

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Background: Gamma irradiation can cause DNA damage in single and double-strand breaks (SSBs & DSBs), especially in peripheral blood lymphocytes. Radiotherapy medical radiation workers can be exposed to gamma radiation related to their daily work. The comet assay is a sensitive method for analyzing DNA damage, especially SSBs. This study explores DNA damage in medical radiation workers’ peripheral blood lymphocytes as an adaptive response using the comet assay.Methods: Blood samples were obtained from four radiotherapy medical radiation workers as a case study (MRW) and two non-medical radiation workers as controls, and then irradiated with various doses of 0, 1, 1.5, and 2 Gy. Lymphocytes were isolated by histopaque and processed by comet assay on the slide under alkaline conditions. The imaging results were analyzed using the Casplab_1.2.3b2 software. The comet assay parameters observed were Tail Length (TL), % Tail DNA (T.DNA), Tail Moment (TM), and Olive Tail Moment (OTM). The one-way ANOVA method was used to analyze statistically between treatment groups. Results: Based on the study results, an increase in TL, T.DNA, TM, and OTM values in all samples was directly proportional to the increase in radiation dose. However, there was no significant difference (P > 0.05) between the MRW group and the control group on each parameter of the comet assay.Conclusions: From this study, it can be concluded that the level of DNA damage of lymphocyte cells as part of the adaptive response in the MRW and control groups was relatively similar after exposure at doses of 0, 1, 1.5, and 2 Gy.

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Assessment of Radiation Induced DNA Damage in Human Peripheral Blood Lymphocytes Using COMET Assay

Cited by 7 publications

References 24 publications

BRCA1 VUS: A functional analysis to differentiate pathogenic from benign variants identified in clinical diagnostic panels for breast cancer

BRCA1 VUS: A functional analysis to differentiate pathogenic from benign variants identified in clinical diagnostic panels for breast cancer

Bilgisayarlı Tomografi İle Yapılan Perkütan Torasik Kitle Biyopsilerinde Lenfositlerdeki DNA Hasarının Değerlendirilmesi

Adaptive Response of Peripheral Blood Lymphocytes in Medical Radiation Workers using the Comet Assay: A Preliminary Studyomet Assay: A Preliminary Study

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