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Updated: Jul 12, 2025

Evaluating In Vitro DNA Damage Using Comet Assay
Published on: October 11, 2017
Comparing P53 expression and genome-wide transcriptome profiling to Comet assay in lymphocytes from melanoma patients
Mojgan Najafzadeh1, Parisa Naeem2, Nader Ghaderi3
1School of Life Sciences, University of Bradford, Richmond Road, Bradford, BD7 1DP, West Yorkshire, UK. m.najafzadeh1@bradford.ac.uk.
Abstract:
This study compared the expression of TP53 in lymphocytes from malignant melanoma (MM) patients with positive sentinel nodes to healthy controls (HCs) following exposure to various doses of UVA radiation. The Lymphocyte Genome Sensitivity (LGS) assay indicated significant differences in DNA damage in lymphocytes between MM patients and HCs. qPCR data demonstrated an overall 3.4-fold increase in TP53 expression in lymphocytes from MM patients compared to healthy controls, following treatment with 0.5 mW/cm2 UVA radiation. Western blotting confirmed that p53 expression was increased in MM lymphocytes following UVA exposure compared to healthy individuals. Genome transcriptome profiling data displayed differences in gene expression between UVA-treated lymphocytes from MM patients and HCs. Peripheral lymphocytes from MM patients are more susceptible to the genotoxic effects of UVA compared to healthy individuals. Our previous studies showed that UVA exposure of various intensities caused significant differences in the levels of DNA damage between lymphocytes from cancer patients compared to HCs through the LGS assay. The present study's results provide further credibility to the LGS assay as a screening test for cancer detection. Peripheral lymphocytes could be a promising blood biopsy biomarker for staging of carcinomas and prevention of carcinoma progression at early stages.

