Assessment of DNA Damage in Some Acute Lymphoblastic Leukemia Iraqi Patients  Using the Comet Assay

Abdul Hussain, Nuria; Hussain Al Faisal, Abdul; Amili, WiaamA.Al

	Assessment of DNA Damage in Some Acute Lymphoblastic Leukemia Iraqi Patients Using the Comet Assay
Al-Ma'mon College Journal
Article 1, Volume 0, Issue 21, 2018, Pages 249-261
Authors
Nuria Abdul Hussain; Abdul Hussain Al Faisal; WiaamA.Al Amili
Abstract
This study investigates genomic damage in peripheral lymphocytes from patients with Acute Lymphoblastic leukemia. As a measure for genomic damage, the comet assay (single-cell gel electrophoresis) was applied. The study was carried out on fifty Iraqi patients ( 34 Male, 16 Female) , aged 2-70 years with Acute Lymphoblastic Leukemia (ALL) . These samples included 20 pretreatment (aged 7-70 years) , 15 relapsed (aged 9-40 years) and 15 under treatment (aged 2-57 years) , compared with 50 apparently healthy normal individuals collected randomly from population living in Baghdad ( aged 3 - 75 years). Patients were treated with nine drugs, which included vincristine , methotrexate, cytosar-U , L-asparaginase , etoposide, , dexamethasone (decadron) , indoxan and steroids. To quantify the DNA damage, three different comet parameters were evaluated , the tail length , % DNA mean in tail and the tail moment .It was found by the comet assay that the tail length (Mean ± SE) for Acute Lymphoblastic Leukemia(ALL) patients (pre treatment, relapse and under treatment the chemotherapy) and controls were 4.77±0.95 px , 20.36±1.86 px ,7.54 ±2.74px and 2.95 ±0. 44px , respectively . There was a significant increase between Acute Lymphoblastic Leukemia patients and controls for mean tail length (P < 0.01).The average damage (percentage of DNA) in the tail region of the comet (Mean ±SE)of Acute Lymphoblastic Leukemia )pre treatment , relapse and post treatment of the chemotherapy) and controls were 0.062 ± 0.036% , 0.121+0.0122% , 0.159±0.067% and 0.017±0.0055%, respectively. There was a significant increase between ALL patients and controls for % DNA mean in tail (P < 0.01). DNA damage were statistically significantly higher (P< 0.01) in case patients (pre treatment, relapse and post treatment of chemotherapy) for tail moment (Mean ± SE)were 0.00040 ± 9.4 E-05, 0.48 ± 0.086 and 0.117 ±0.054, respectively than in control subjects 0.000142 ± 4.6E-05. In addition, the results of comet assay are affected by for gender when compared with the groups studied . Despite their limitations, present results confirm the usefulness of the comet assay as a sensitive biomarker of exposure that enables rapid and simple detection of primary DNA damage in peripheral blood leukocytes of ALL patients. The comet assay provides a powerful technique for the routine detection of critical DNA lesions produced after the administration of antineoplastic drugs in clinical settings. These results , emphasize the need to further optimize the current therapy for reducing the degree of genomic damage.
Keywords
Comet Assay; Human Lymphocytes; Chemotherapy; Acute Lymphocyte Leukemia

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