The comet assay: what can it really tell us?

Collins, Andrew; Dobson, V L; Dušinská, Maria; Kennedy, Gayle; Štětina, Rudolf

doi:10.1016/s0027-5107(97)00013-4

Cited by 601 publications

(321 citation statements)

References 16 publications

Supporting

Mentioning

304

Contrasting

Unclassified

Order By: Relevance

“…Recognition and incision are the rate-limiting steps in NER while rejoining of DNA strand breaks is a rapid process with a halftime of a few minutes in most cell types [41]. DNA breaks are only transiently present in the NER process [42]. In our experiments, after BPDE treatment, the peripheral blood lymphocytes were incubated for an additional 24 hours before the comet assay was performed.…”

Section: Discussionmentioning

confidence: 98%

Modulation of DNA damage/DNA repair capacity by XPC polymorphisms

et al. 2008

View full text Add to dashboard Cite

XPC, a key protein in the nucleotide excision repair (NER) pathway, recognizes damaged DNA and initiates NER. Genetic variations in the XPC gene might be associated with altered DNA repair capacities (DRC). In this study, we genotyped three XPC polymorphisms, Ala499Val (C→T), PAT (-/+) and Lys939Gln (A→C), and measured the DNA damage/DRC by alkaline comet assay challenged by BPDE and gamma-radiation in 476 healthy subjects. We also evaluated the associations between DNA damage/DRC and genotypes of XPC polymorphisms. Compared with the XPC Lys939Gln homozygous wild type (AA) subjects, subjects with the variant alleles (AC and CC) had significantly higher DNA damages induced by BPDE (Median and 95% confidence interval [CI]: 3.16 (3.01-3.44) vs. 2.88 (2.51-3.05), P=0.01), and gamma-radiation (4.18(3.94-4.44) vs. 3.71 (3.49-4.04), P=0.01). However, subjects with the variant alleles (CT and TT) of Ala499Val exhibited a 8.6 % and 13.1% decrease in DNA damages induced by BPDE (P=0.05) and gamma-radiation (P=0.001), respectively. Significant correlations were found between genotypes and induced DNA damages in XPC Lys939Gln (For BPDE: R=0.12, P=0.01; for gamma-radiation: R=0.094, P=0.046) and Ala499Val (For BPDE: R=-0.11, P=0.03; for gamma-radiation: R=-0.16, P=0.0009). The haplotypes "T-A" (in the order of Ala499Val-PAT-Lys939Gln) was associated with the lowest DNA damages. Our results suggested that the DRC of host cells might be modulated by specific XPC polymorphisms.

show abstract

Section: Discussionmentioning

confidence: 98%

Modulation of DNA damage/DNA repair capacity by XPC polymorphisms

et al. 2008

View full text Add to dashboard Cite

show abstract

“…However, many forms of structural damage can be converted to strand breaks during cell preparation and electrophoresis (Collins et al, 1997;Olive, 1999) . In this study, we report on the increased DNA strand breaks in Hela cells after exposure to goniothalamin.…”

Section: Discussionmentioning

confidence: 99%

“…Different scores indicate different degrees of DNA damage was described (Collins, 1997). Collins et al have described that at score 2, the proportion of DNA at the tail is between 25 and 75% and at score 3, more than 75% of DNA is at the tail.…”

Section: Discussionmentioning

confidence: 99%

“…The Comet assay was performed under alkaline conditions using the method adapted from Singh et al (1980) as described previously by Gedik et al (1992) and Collins et al (1997). In brief, Hela cells with goniothalamin treatment were suspended in 1% low melting point agarose in PBS, pH 7.4, at 37°C, and 100 μl were pipetted onto a frosted glass microscope slide pre-coated with an 100 μl layer of 1% normal melting point agarose.…”

Section: Apoptosis In Cancer Cells Treated With Goniothalamin Extractmentioning

confidence: 99%

See 1 more Smart Citation

Induction of Caspase-9, Biochemical Assessment and Morphological Changes Caused by Apoptosis in Cancer Cells Treated with Goniothalamin Extracted from Goniothalamus macrophyllus

Alabsi¹,

Ali²,

Ali³

et al. 2013

Asian Pacific Journal of Cancer Prevention

View full text Add to dashboard Cite

“…The comet assay is a widely used method for studying DNA damage (Collins et al, 1997). The main advantages of this procedure as compared to other methods, such as alkaline unwinding, alkaline elution, or HPLC detection of oxidized DNA bases, are the high sensitivity (less than 0.5 breaks/10 9 Da of DNA can be detected) and the single-cell resolution.…”

Section: Discussionmentioning

confidence: 99%

Loss of tyrosinase activity confers increased skin tumor susceptibility in mice

et al. 2004

View full text Add to dashboard Cite

The tyrosinase (Tyr) gene encodes the enzyme tyrosinase that catalyses the conversion of L-tyrosine into DOPA (3,4-dihydroxyphenylalanine)-quinone. The albino mutation abrogates functional activity of tyrosinase resulting in deficiency of melanin pigment production in skin and retina. Tyr maps to a region in the central position of Chromosome 7 that contains a skin tumor-modifier locus. We rescued the albino mutation in transgenic mice to assess a possible role of Tyr gene in two-stage skin carcinogenesis. Transgenic expression of the functional Tyr Cys allele in albino mice (Tyr Ser ) caused a reduction in skin papilloma multiplicity, in four independent experiments and at three dose levels of DMBA (9,10-dimethyl-1,2-benzanthracene). In vitro mechanistic studies demonstrated that transfection of the Tyr Cys allele in a human squamous cell carcinoma cell line (NCI-H520) increases tyrosinase enzyme activity and confers resistance to hydrogen peroxide-induced oxidative DNA damage. These results provide direct evidence that the Tyr gene can act as a skin cancer-modifier gene, whose mechanism of action may involve modulation of oxidative DNA damage.

show abstract

The comet assay: what can it really tell us?

Cited by 601 publications

References 16 publications

Modulation of DNA damage/DNA repair capacity by XPC polymorphisms

Modulation of DNA damage/DNA repair capacity by XPC polymorphisms

Induction of Caspase-9, Biochemical Assessment and Morphological Changes Caused by Apoptosis in Cancer Cells Treated with Goniothalamin Extracted from Goniothalamus macrophyllus

Loss of tyrosinase activity confers increased skin tumor susceptibility in mice

Contact Info

Product

Resources

About