scholarly journals SMOKING GENOTOXICITY IN PERIPHERAL BLOOD LYMPHOCYTES USING THE COMET ASSAY

2013 ◽  
Vol 03 (03) ◽  
pp. 038-041
Author(s):  
Shobha S. Shetty ◽  
Hrishikesh Nachane

Abstract Background: Smoking has been shown to have a positive effect on DNA damage in almost all the cells of the body. Quantitative analysis of this damage will help in assessing the etiopathogenesis of various nicotine induced damage to the body. Comet assay has been an emerging tool in this regard and hence was applied by us to estimate the severity of DNA damage in smokers. Aims & Objectives: To evaluate the DNA genotoxicity in peripheral blood lymphocytes in smokers and their comparison with non smokers & assess the quantitative damage. Materials and methods: 30 smokers & 20 non smokers were recruited & their peripheral blood was taken for the comet assay to look for Olive moment & Tail moment to quantitatively assess the DNA damage due to cigarette smoking. Results: In our study there was no significant difference in the analysis of DNA damage (with regard to tail moment & olive moment) in smokers versus non smokers (P value: more than 0.05). Conclusions: Though smoking is known to cause DNA damage, we did not find significant differences between the two groups probably due to other multifactorial etiologies for genotoxicity.

2021 ◽  
Vol 8 (1) ◽  
pp. 105-113
Author(s):  
Darlina Yusuf ◽  
Devita Tetriana ◽  
Tur Rahardjo ◽  
Teja Kisnanto ◽  
Yanti Lusiyanti ◽  
...  

Analyses of DNA Damage in the Patient’s Lymphocyte Cells Post-Radiotherapy Radiotherapy given in high doses to kill cancer cells can also induce DNA damage in surrounding normal cells. The radiation dose is divided into smaller doses called fractionation to decrease the effect of radiation on normal tissue. For this reason, it is necessary to monitor the peripheral blood lymphocytes to evaluate the patient's DNA damage. The alkaline comet test is a simple and sensitive technique for detecting DNA instability. This study involved 11 patients who underwent radiotherapy up to 20 Gy, and 11 healthy subjects as controls. This study aims to see how much DNA damage is caused by a 20 Gy fractionated radiation dose in patients with various cancers. The results showed that the mean frequency of damaged cells in patients was 80.54 ± 12.52% with a mean comet tail length of 49.98 ± 12.93 µm. There was a significant difference in both the frequency of damaged cells and the mean value of the comet tail length against the control group (p < 0.001). It was concluded that high doses of radiation can cause DNA damage to peripheral blood lymphocytes. Radioterapi yang diberikan dalam dosis tinggi untuk mematikan sel kanker juga dapat menginduksi kerusakan DNA pada sel normal di sekitarnya. Dosis radiasi dibagi menjadi dosis yang lebih kecil yang disebut fraksinasi untuk menurunkan efek radiasi pada jaringan normal. Untuk itu perlu pemantauan pada limfosit darah tepi untuk mengevaluasi kerusakan DNA pasien. Uji komet alkali merupakan teknik yang sederhana dan sensitif untuk mendeteksi ketidakstabilan DNA. Penelitian ini melibatkan 11 pasien yang menjalani radioterapi hingga 20 Gy, dan 11 subyek sehat sebagai kontrol. Penelitian ini bertujuan untuk melihat seberapa besar kerusakan DNA akibat dosis radiasi fraksinasi 20 Gy pada pasien dengan variasi kanker. Hasil penelitian menunjukkan bahwa rerata frekuensi sel yang rusak pada pasien 80,54 ± 12,52% dengan rerata panjang ekor komet 49,98 ± 12,93 µm terdapat perbedaan nyata baik pada frekuensi sel yang rusak maupun nilai rerata panjang ekor komet terhadap kelompok kontrol (p < 0,001). Penelitian ini menyimpulkan bahwa radiasi dosis tinggi dapat menyebabkan kerusakan DNA sel limfosit darah tepi.


2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 10641-10641 ◽  
Author(s):  
F. Franke ◽  
M. Agnoletto ◽  
J. Saffi ◽  
T. Guecheva

10641 Breast cancer is the most common malignancy among women and its rate of mortality is still high. The increase knowledge of breast cancer biology is heaving great impact on determining the clinical prognosis and response to treatment. Impaired DNA repair may elevate the risk of malignant transformation of breast cells due to the accumulation of spontaneous mutations in target genes and increasing susceptibility to exogenous carcinogens. The present study was designed to evaluate the relationship between DNA damage and expression of some critical genes including TP53, c-ERBB2, ER (Estrogen Receptor) and PR (Progesterone Receptor) in breast cancer. Blood samples were obtained from female patients with diagnosed breast cancer before chemotherapy as well as from healthy individuals, and were processed in 24 hours. To evaluate the role of DNA repair in breast cancer we determined the level of DNA damage and the capacity to remove DNA damage induced by hydrogen peroxide in the peripheral blood lymphocytes. For this purpose the alkaline version of the comet assay, which provides a sensitive tool to investigate DNA damage and repair, was applied. The level of basal DNA damage was higher in breast cancer patients compared to the control group. Considerable inter-individual variations of DNA damage and repair in breast cancer patients were observed both before and after the treatment. The correlation between DNA damage in peripheral blood and expression of p53, c-erbB-2, PR and ER was analyzed. This preliminary study indicates that the DNA damage accumulation, observed in peripheral blood lymphocytes of breast cancer patients in early stages, could be attributed to impaired DNA repair. Our results suggest that DNA damage, as evaluated by the comet assay, seems to be useful molecular biomarker for monitoring ongoing exposures to DNA damaging agents. Such a research on the mutagen sensitivity and efficacy of DNA repair could impact on the development of new diagnostic and screening strategies. Work Supported by FAPERGS and GENOTOX (UFRGS). No significant financial relationships to disclose.


2015 ◽  
Vol 66 (2) ◽  
pp. 153-158 ◽  
Author(s):  
Fulya Dilek Gökalp Muranli ◽  
Martin Kanev ◽  
Kezban Ozdemir

Abstract The aim of this study was to evaluate the genetic damage in human peripheral blood lymphocytes following 24 and 48- hour exposure to a commercial diazinon formulation Basudin 60EM® at concentrations between 0.01 and 40 μg mL-1. For this purpose we used the micronucleus (MN), fluorescence in situ hybridization (FISH), and alkaline single cell gel electrophoresis (comet) assay. Diazinon significantly increased the frequency of micronucleated cells compared to control. Forty-eight-hour exposure increased this frequency even at lower concentrations (0.01-10 μg mL-1). The FISH results revealed aneugenic effects at 10 μg mL-1. The comet assay also confirmed DNA damage at concentrations between 10 and 40 μg mL-1. Our findings have confirmed the genotoxic potential of diazinon and its cytotoxic effect on human lymphocytes. The increased DNA damage in our study raises concern about the current assessment of the health risk posed by this pesticide and calls for a high level of caution in agricultural and household use.


Author(s):  
Anita Yadav ◽  
Surbhi Bal ◽  
Neha Verma ◽  
Neeraj K. Aggarwal ◽  
Ranjan Gupta

Arsenic contamination is one of the major health concerns all over the world and associated with various types of cancer and pathological effects. The production of reactive oxygen species (ROS) plays a crucial role in arsenic mediated toxicity. Several studies have shown that population constantly exposed to arsenic have substantial oxidative stress that, in turn, induces DNA damage. In the present work eugenol and anethole were investigated for their protective effect against arsenic mediated oxidative DNA damage in peripheral blood lymphocytes. Comet assay and lipid peroxidation was used as biomarker of genotoxicity and oxidative stress respectively. A dose dependent increase in tail moment and lipid peroxidation was observed when lymphocytes were treated with sodium arsenite. Treatment of arsenic (50?M) along with eugenol (20?M) and anethole (50?M) showed a significant decrease in the tail moment and lipid peroxidation in cultured peripheral blood lymphocytes. The decrease in the tail moment and lipid peroxidation was significantly higher in combined supplementation of eugenol and anethole as compared to individual administration. The results of the present study suggests ameliorative role of eugenol and anethole against arsenic induced genotoxic and oxidative damage in cultured human peripheral blood lymphocytes.


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