Single cell gel electrophoresis assay: a new technique for human biomonitoring studies

Kassie, Fekadu; Parzefall, Wolfram; Knasmüller, Siegfried

doi:10.1016/s1383-5742(00)00041-7

Cited by 329 publications

(158 citation statements)

References 59 publications

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“…In general, strand breaks result from the attack of a variety of endogenous and exogenous factors to the sugar residues of DNA (21). SSBs represent the initial DNA damage and are often used as a biomarker of exposure (22)(23)(24). DSBs are considered to be more biologically relevant since they can lead to chromosome INTERNATIONAL JOURNAL OF ONCOLOGY 35: 1297-1304, 2009 Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells translocation and cancer if repaired improperly in addition to genetic instability and cell death if left unrepaired (25).…”

Section: Introductionmentioning

confidence: 99%

Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells

Moktar

Ravoori²,

Vadhanam³

et al. 2009

Int J Oncol

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Abstract. Human papillomavirus (HPV) is the causative factor in the development and progression of cervical cancers in >97% of the cases, although insufficient. Epidemiological studies suggest an elevated risk of cervical cancer for cigarette smokers; therefore, we examined cigarette smoke-induced DNA damage and repair in HPV16-transformed human ectocervical cells (ECT1/E6 E7). Cells were treated with cigarette smoke condensate (CSC) for 72 h to assess the formation of single-and double-strand DNA breaks, measured by alkaline and neutral single cell gel electrophoresis assays, respectively. The mean tail length of cells with single-strand breaks was increased by 1.8-, 2.7-and 3.7-fold (p<0.001) after treatment with 4, 8 and 12 μg/ml CSC, respectively. The tail length with double-strand breaks was also increased dose-dependently. These results were further supported by measurement of the mean tail moment: the increase in both single-and double-strand breaks were much more pronounced with increasing concentration of CSC, by up to 23.5-fold (p<0.0001 for both assays). To examine the DNA repair, cells were treated with CSC for 72 h, followed by CSC withdrawal and re-incubation of the cells with fresh medium for 24, 48, or 72 h. Both single-and double-strand DNA breaks were removed during the initial 24 h but no further removal of the damage was observed. Up to 80% of residual single-and double-strand DNA breaks (p<0.05) were found to persist at all CSC concentrations examined. Ellagic acid, a known antioxidant and free-radical scavenger, was found to significantly inhibit DNA breaks induced by CSC. Thus, free radicals may be a plausible source of CSCinduced DNA damage. These data show that CSC-mediated DNA strand breaks are highly persistent, and suggest that persistence of cigarette smoke-associated DNA damage in the presence of HPV infection may lead to increased mutations in cervical cells and ultimately higher cancer risk.

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Section: Introductionmentioning

confidence: 99%

Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells

Moktar

Ravoori²,

Vadhanam³

et al. 2009

Int J Oncol

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“…The PBMCs consisting of lymphocytes are commonly chosen for conducting cytotoxicity and genotoxicity testing of plant extracts because they are vulnerable to free radicals, which results in damaging effects (Heaton et al 2002). Furthermore, the cells are considered a suitable indicator of the actual state of the body (Kassie et al 2000).…”

Section: Cytotoxic Effects Of the Plant Extractsmentioning

confidence: 99%

Chemical Constituents and Toxicity Screening of Three Aromatic Plant Species from Peninsular Malaysia

Jumaat¹,

Tajuddin²,

Sudmoon³

et al. 2017

BioResources

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Medicinal and aromatic plants (MAPs) are widely valued for their aromas, tastes, and treatments for various human illnesses. The chemical constituents and toxicity content of three aromatic plant species, Syzygium polyanthum Wight (Walp.), Monocarpia marginalis (Scheff.) J. Sinclair, and Chromolaena odorata (L.) R.M. King & H. Rob, were determined, via gas chromatography (GC) with mass spectrometry (MS), and flame ionization detector (FID). Altogether, 116 compounds were identified in the essential oils and hexane and methanol extracts. Toxicity evaluations were carried out on human peripheral blood mononuclear cells (PBMCs). Three plant samples were found toxic to human PBMCs. The essential oils of M. marginalis and C. odorata, and the hexane extract of C. odorata yielded IC50 and LD50 values of 76 mg/mL and 6,913 mg/kg, 14 mg/mL and 3,684 mg/kg, and 2.45 mg/mL and 1,927 mg/kg, respectively. Based on the LD50 values, M. marginalis and C. odorata can be classified as slightly and moderately hazardous, respectively. A detailed toxicity evaluation via comet assay showed that M. marginalis and C. odorata induced significant DNA damage (p < 0.05). As for S. polyanthum, the species did not give any cytotoxic or genotoxic evidences.

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“…Alkaline Comet assay is a widely-used, rapid, and sensitive technique for the measurement of DNA damage both in vitro and in vivo (30)(31)(32). In the present study, we applied this technique for the evaluation of DNA damaging potential of the extract.…”

Section: Alkaline Comet Assaymentioning

confidence: 99%

Investigation of Cytotoxic and Genotoxic Potential of Cinnamomum cassia Bark Water Extract

Karadağlı¹,

Agrap²,

Erciyas

2014

MUSBED

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ÖZETCinnamomum cassia kabuğu su ekstresinin sitotoksik ve genotoksik potansiyelinin araştırılması Amaç: Son zamanlarda, tarçın ile ilgili çalışmalar zengin polifenol içeriğine bağlı antioksidan aktivitesi üzerinde yoğunlaşmıştır. Polifenollerin aynı zamanda pro-oksidan olarak hareket ettikleri ve DNA'da oksidatif zincir kırıklarına neden oldukları bildirilmiştir. Bu çalışmada Cinnamomum cassia su ekstresinin insan periferal kan lenfositlerindeki sitotoksik ve genotoksik potansiyelini araştırdık. Yöntemler: Cinnamomum cassia su ekstresi, toz haline getirilmiş tarçın kabuğunun ultra saf suda 72 saat maserasyonu ile hazırlandı. Ekstrenin lenfositlerdeki sitotoksik etkisi WST-1 yöntemi ile araştırıldı. Ekstrenin DNA'da hasar oluşturucu potansiyelini değerlendirmek için alkali Komet yöntemi uygulandı. DNA hasarı kuyruk DNA yüzdesi ve kuyruk momenti şeklinde ifade edildi. Bulgular: Lenfositlerin canlılığı ekstre ile 24 saat muamele sonrası konsantrasyona bağımlı olarak azaldı. Ekstre ≥400 µg/ml konsantrasyonlarda negatif kontrole göre anlamlı düzeyde DNA hasarına neden oldu. Sonuç: Çalışma sonuçlarımız Cinnamomum cassia kabuğu su ekstresinin in vitro olarak sitotoksik ve genotoksik etki potansiyelini göstermektedir. Ekstrenin özellikle yüksek dozlarda ya da uzun süreli kullanımında güvenliliği açısından bu bulgu önemli görünmektedir ve bu nedenle in vivo çalışmalarla aydınlatılmalıdır. Anahtar sözcükler: Cinnamomum cassia, alkali Komet yöntemi, sitotoksisite, pro-oksidan aktivite ABS TRACT Investigation of cytotoxic and genotoxic potential of cinnamomum cassia bark water extractObjective: Recently many investigations on cinnamon have focused on its powerful antioxidant activity due to its rich polyphenol content. Polyphenols have also been reported to act as pro-oxidants, causing oxidative strand breaks in DNA. In the present study, we investigated the cytotoxic and genotoxic potential of Cinnamomum cassia water extract in human peripheral blood lymphocytes. Methods: Cinnamomum cassia water extract was prepared from grounded bark of cinnamon by maceration with ultrapure water for 72 h. Cytotoxicity of the extract on lymphocytes was determined by WST-1 assay. Alkaline Comet assay was conducted to evaluate the DNA damaging potential of the extract. DNA damage was expressed as DNA percentage in the tail and tail moment. Results: The viability of lymphocytes was decreased by treatment with the extract for 24 h, as a concentration-dependent manner. At the concentrations ≥400 µg/ml, the extract induced significant DNA damage compared to negative control. Conclusion: Our results show the cytotoxic and genotoxic potential of Cinnamomum cassia bark water extract in vitro. This finding seems a significant safety concern, particularly in high doses or long term use of the extract, and therefore needs to be clarified by in vivo studies.

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Single cell gel electrophoresis assay: a new technique for human biomonitoring studies

Cited by 329 publications

References 59 publications

Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells

Cigarette smoke-induced DNA damage and repair detected by the comet assay in HPV-transformed cervical cells

Chemical Constituents and Toxicity Screening of Three Aromatic Plant Species from Peninsular Malaysia

Investigation of Cytotoxic and Genotoxic Potential of Cinnamomum cassia Bark Water Extract

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