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Link to original content: http://pubmed.ncbi.nlm.nih.gov/23093842/
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. 2012:2012:385245.
doi: 10.1155/2012/385245. Epub 2012 Oct 3.

Evaluation of DNA single and double strand breaks in women with cervical neoplasia based on alkaline and neutral comet assay techniques

Elva I Cortés-Gutiérrez  1 , Fernando Hernández-Garza, Jorge O García-Pérez, Martha I Dávila-Rodríguez, Miguel E Aguado-Barrera, Ricardo M Cerda-Flores
Affiliations

Evaluation of DNA single and double strand breaks in women with cervical neoplasia based on alkaline and neutral comet assay techniques

Elva I Cortés-Gutiérrez et al. J Biomed Biotechnol. 2012.

Abstract

A hospital-based unmatched case-control study was performed in order to determine the relation of DNA single (ssb) and double (dsb) strand breaks in women with and without cervical neoplasia. Cervical epithelial cells of 30 women: 10 with low grade squamous intraepithelial lesions (LG-SIL), 10 with high-grade SIL (HG-SIL), and 10 without cervical lesions were evaluated using alkaline and neutral comet assays. A significant increase in global DNA damage (ssb + dsb) and dsb was observed in patients with HG-SIL (48.90 ± 12.87 and 23.50 ± 13.91), patients with LG-SIL (33.60 ± 14.96 and 11.20 ± 5.71), and controls (21.70 ± 11.87 and 5.30 ± 5.38; resp.). Pearson correlation coefficient reveled a strong relation between the levels ssb and dsb (r(2) = 0.99, P = 0.03, and r(2) = 0.94, P = 0.16, resp.) and progression of neoplasia. The increase of dsb damage in patients with HG-SIL was confirmed by DNA breakage detection-FISH (DBD-FISH) on neutral comets. Our results argue in favor of a real genomic instability in women with cervical neoplasia, which was strengthened by our finding of a higher proportion of DNA dsb.

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Figures

Figure 1
Figure 1
Classification of the comets. Normal nucleus (class 0), halo around the nucleus (class 1), and gradual increase in the length of the comet tail, evolving in parallel with a decrease in the nuclear DNA content (class 2 and class 3). Alkaline comet assay reflecting global DNA damage (ssb + dsb) (upper panel), and neutral comet assay reflecting dsb mainly (lower panel).
Figure 2
Figure 2
Epithelial cells of the cervix processed using DBD-FISH on neutral comet to determine dsb and ssb simultaneously using whole-genome-labeled (red) and DAPI counterstained (blue) cells. Cell of patients with HG-SIL (c) and exhibit an increase in the migration length (dsb) and in the fluorescence intensity (ssb) compared with cells from control women ((a) and (b)). Apoptotic cell was concluded to form comets with large fan-like tails and small heads (i.e., so-called hedgehogs) (d).
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