The comet assay: A sensitive method for detecting DNA damage in individual cells

Liao, Wenjuan; McNutt, Michael A.; Zhu, Wei‐Guo

doi:10.1016/j.ymeth.2009.02.016

Cited by 282 publications

(179 citation statements)

References 38 publications

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“…The genome-protective eff ect of xanthohumol against oxidative DNA damage induced by H 2 O 2 was evaluated using the yeast comet assay. The yeast comet assay is a well-established, simple, versatile, sensitive and inexpensive technique used to assess DNA oxidative damage quantitatively and qualitatively in eukaryotic cell populations (30)(31)(32)(33). Yeast spheroplasts were incubated with diff erent xanthohumol concentrations and 10 mM H 2 O 2 in S buff er (containing 1 M sorbitol) in order to maintain osmotic protection.…”

Section: Xanthohumol Protects Yeast Cells From Dna Damage Induced Bymentioning

confidence: 99%

Dose-Dependent Protective and Inductive Effects of Xanthohumol on Oxidative DNA Damage in Saccharomyces cerevisiae

Carvalho

Oliveira

Johansson

et al. 2016

Food Technol. Biotechnol.

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SummaryThe eff ect of xanthohumol, a prenylfl avonoid isolated from the hop plant (Humulus lupulus L.), on Saccharomyces cerevisiae DNA oxidative damage and viability was evaluated. Yeast cultures under oxidative stress, induced by H 2 O 2 , displayed stronger growth in the presence of 5 mg/L of xanthohumol than cultures with only H 2 O 2 . Likewise, DNA damage assessed by the comet assay was signifi cantly lower in cells co-incubated with xanthohumol and H 2 O 2 . Accordingly, fl uorescence of dichlorofl uorescein in cells treated with H 2 O 2 and xanthohumol was considerably lower than in cells exclusively treated with H 2 O 2 , indicative of a reactive oxygen species scavenging mechanism and consequent formation of oxidation products, as detected by mass spectrometry. However, at concentrations above 5 mg/L, xanthohumol elicited an opposite eff ect, leading to a slower growth rate and significant increase in DNA damage. A yeast yap1 deletion mutant strain sensitive to oxidative stress grew more slowly in the presence of at least 5 mg/L of xanthohumol than cultures of the wild type, suggesting that xanthohumol toxicity is mediated by oxidative stress. This evidence provides further insight into the impact of xanthohumol on yeast cells, supporting dose-dependent antioxidant/antigenotoxic and prooxidant/genotoxic eff ects.

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Section: Xanthohumol Protects Yeast Cells From Dna Damage Induced Bymentioning

confidence: 99%

Dose-Dependent Protective and Inductive Effects of Xanthohumol on Oxidative DNA Damage in Saccharomyces cerevisiae

Carvalho

Oliveira

Johansson

et al. 2016

Food Technol. Biotechnol.

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“…We also measured the formation of DNA strand breaks in L3MBTL1-depleted cells using the comet assay, which visualizes damaged DNA on a single-cell level (15). Comet tails were detected in depleted U2OS cells (Fig.…”

Section: Depletion Of L3mbtl1 Inhibits Cell Proliferation and Causesmentioning

confidence: 99%

L3MBTL1 polycomb protein, a candidate tumor suppressor in del(20q12) myeloid disorders, is essential for genome stability

Gurvich¹,

Perna²,

Farina

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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The l3mbtl1 gene is located on the long arm of chromosome 20 (q12), within a region commonly deleted in several myeloid malignancies. L3MBTL1 is a human homolog of the Drosophila polycomb L(3)MBT tumor suppressor protein and thus a candidate tumor suppressor in del(20q12) myeloid disorders. We used the loss-of-function approach to explore the possible tumor suppressive mechanism of L3MBTL1 and found that depletion of L3MBTL1 from human cells causes replicative stress, DNA breaks, activation of the DNA damage response, and genomic instability. L3MBTL1 interacts with Cdc45, MCM2-7 and PCNA, components of the DNA replication machinery, and is required for normal replication fork progression, suggesting that L3MBTL1 causes DNA damage, at least in part, by perturbing DNA replication. An activated DNA damage response and genomic instability are common features in tumorigenesis and a consequence of overexpression of many oncogenes. We propose that the loss of L3MBTL1 contributes to the development of 20q − hematopoietic malignancies by inducing replicative stress, DNA damage, and genomic instability. T he l3mbtl1 gene is located on the long arm of chromosome 20q, within a region on 20q12 commonly deleted in several myeloid malignancies, including myeloproliferative neoplasms, myelodysplastic syndromes, and acute myeloid leukemia (1). It has been proposed that the 20q12 locus contains one or more tumor suppressors, which when lost contribute to the development of these disorders. L3MBTL1 is a human homolog of the Drosophila polycomb group (PcG) protein L(3)MBT. Homozygous mutations of the Drosophila l3mbt gene cause malignant transformation of the adult optic neuroblasts and ganglion mother cells in the larval brain (2). Somatic, focal deletions of other human L3MBTL family members, the l3mbtl2 and l3mbtl3 genes, have recently been found in human medulloblastoma (3). These findings suggest that L3MBTL1 is a candidate tumor suppressor gene in myeloid malignancies associated with 20q12 deletions.The L3MBTL1 protein contains three MBT repeats, which assume a three-bladed propeller-like architecture, as well as a Zn finger and an SPM dimerization domain (4, 5). We previously demonstrated that L3MBTL1 functions as an HDAC-independent transcriptional repressor (6) that binds preferentially to mono-and dimethylated lysines of histones via the second of its three MBT repeats (7,8). The three MBT domains of L3MBTL1 are sufficient to compact nucleosomal arrays. This compaction requires that the nucleosome contain a mono-or dimethylated lysine 26 on histone H1b or lysine 20 on histone H4 (H4K20) (7). L3MBTL1 binds to chromatin most prominently during the S phase of the cell cycle, concomitant with the appearance of the monomethylated H4K20 (H4K20me1) mark (8), suggesting that the biological function of L3MBTL1 may be related to DNA replication.The accurate duplication of DNA during replication is essential for maintaining genomic stability, as uncorrected errors made during this process can lead to DNA breaks, which generate mutat...

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“…9 Kadar MDA plasma diukur dengan metode thiobarbituric acid reactive substance (TBARS) menggunakan spektrofotometri. 11 Karena MDA bersifat tidak stabil, digunakan larutan standar tetraetoksipropan (TEP). Jika MDA dreaksikan dengan TBA akan membentuk larutan berwarna merah muda dan menyerap cahaya pada panjang gelombang 530 nm.…”

Section: Gambar 2 Pengukuran Panjang Nasoanalunclassified

Potensi Suplementasi Resveratrol dalam Mengatasi Efek Negatif Hormon Tiroid pada Terapi Obesitas: Uji Preklinik

Ruslami¹,

Rosalita²,

Yolanda³

et al. 2016

mkb

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AbstrakTerapi obesitas menggunakan hormon tiroid menurunkan berat badan dengan nyata, namun menyebabkan berbagai efek negatif seperti stres oksidatif, kerusakan DNA, dan muscle wasting. Penelitian ini bertujuan mengkesplorasi potensi resveratrol (RSV), polifenol alami yang mempunyai efek antioksidan, anti-inflamasi, dan antiobesitas. Uji preklinik dengan rancangan acak sederhana dilakukan di Lab Hewan Fakultas Kedokteran Universitas Padjadjaran menggunakan 40 ekor tikus Wistar yang diinduksi obesitas. Hewan model obesitas dialokasikan secara acak ke dalam dua kelompok selama periode Agustus-November 2014. Kelompok perlakuan mendapat hormon tiroid 3 (HT3) dan RSV, serta kelompok kontrol mendapat HT3 saja selama 14 hari. Hewan coba yang mendapat HT3 dan RSV memiliki bobot badan lebih besar (290,5 g vs 251 g, p=0,016), massa otot lebih berat (2,57 g vs 2,25 g, p=0,036), indeks adiposit lebih kecil (0,89% vs 1,02%, p=0,026), dan area under the curve (AUC) gukosa lebih kecil (1.849,2 mg.mnt/dL vs 2.591,7 mg.mnt/dL, p<0,001) dibanding yang mendapat HT3 saja. Tidak terdapat perbedaan kadar MDA plasma (0,49 nmol/mL vs 0,44 nmol/mL, p=0,430) dan kerusakan DNA (19,7 pixel vs 22,0 pixel, p=0,657) pada kedua kelompok yang mungkin berhubungan dengan dosis dan lama pemberian resveratrol. Suplementasi resveratrol berpotensi mengatasi efek negatif hormon tiroid pada hewan model obesitas. Perlu penelitian lanjutan untuk mengetahui dosis optimal resveratrol sebagai antioksidan pada pengobatan obesitas menggunakan hormon tiroid. Treatment of obesity using thyroid hormone could give significant result in decreasing body weight, but it also causes various negative effects such as oxidative stress, DNA damage, and muscle wasting. This sudy aimed to explore the potential of resveratrol, a natural polyphenol, that has anti-oxidant, anti-inflammatory, and anti-obesity effects, in managing negative effects of thyroid hormone used as anti-obesity. A preclinical trial was conducted at the animal laboratory, Faculty of Medicine, Universitas Padjadjaran. Forty male Wistar rats underwent obesity induction before randomly allocated into intervention and control groups in August-November 2014. The intervention group received a 14-days of triiodothyronine (T3) and resveratrol (RSV), while the control group received T3 only. Obese rats treated with T3 and RSV were heavier (body weight of 290.5 g vs 251.0 g, p<0.016), had a heavier muscle mass (2.57 g vs 2.25 g, p=0.036), smaller adipocyte index, (0.89% vs 1.02%, p=0.026) and smaller glucose area under the curve (AUC) (1,848.2 mg.mnt/dL vs 2,591.7 mg.mnt/dL, p<0.001) than those who were treated with T3 only. There was no significance difference in MDA plasma levels (0.49 nmol/mL vs 0.44 nmol/mL, p=0.0430) and DNA damage (19.7 pixel vs 22.0 pixel, p=0.657) between groups, probably due to insufficient dosage of resveratrol. Our results indicate that resveratrol supplementation could counter negative effects of thyroid hormone used as anti-obesity in obese-rats model treated with T3. More research is...

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The comet assay: A sensitive method for detecting DNA damage in individual cells

Cited by 282 publications

References 38 publications

Dose-Dependent Protective and Inductive Effects of Xanthohumol on Oxidative DNA Damage in Saccharomyces cerevisiae

Dose-Dependent Protective and Inductive Effects of Xanthohumol on Oxidative DNA Damage in Saccharomyces cerevisiae

L3MBTL1 polycomb protein, a candidate tumor suppressor in del(20q12) myeloid disorders, is essential for genome stability

Potensi Suplementasi Resveratrol dalam Mengatasi Efek Negatif Hormon Tiroid pada Terapi Obesitas: Uji Preklinik

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