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Showing 2 results for Comet Assay
Bahare Nikoozar , Negin Kazemi , Abbas Kiani-Esfahani , Mohammad Hossein Nasr-Esfahani , Marziyeh Tavalaee ,
Volume 25, Issue 3 (10-2023)
Abstract
One of the main spermatogenesis events is the replacement of histones with small proteins called protamines, which leads to chromatin's condensation in the sperm nucleus and protects it against possible damage. Today, tests such as aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining are used based on different characteristics to evaluate sperm chromatin compaction. For the assessment of DNA fragmentation in sperm, several tests such as 8-hydroxy-2-deoxyguanosine (8-OHdG), TUNEL, Comet, sperm chromatin structure assay (SCSA), sperm chromatic dispersion (SCD), and acridine orange have been introduced that directly and indirectly assay DNA damage. The articles in PubMed and Google Scholar, as well as related books, from 2007 to 2022, were collected and reviewed based on keywords 8-OHdG, TUNEL, Comet, SCD, and acridine orange. So far, many studies have been conducted at the treatment level and on sperm chromatin tests, but the number of cases published so far is limited. Various sperm samples have been used in different studies, with different threshold limits in the tests. The sixth edition of the World Health Organization (WHO) book notes that each laboratory has its threshold limit. Therefore, in this review study, common methods of evaluating chromatin packaging and DNA damage are introduced, and the advantages and disadvantages of each test are discussed based on the latest achievements related to infertility.
Elahe Gharehkhani , Sajedeh Zibayi, Mahboube Rahmati Kukandeh , Ramin Ataee , Mohammad Shokrzadeh ,
Volume 27, Issue 4 (12-2025)
Abstract
Background and Objective: 5-fluorouracil (5-FU) is a widely used chemotherapeutic agent for colorectal cancer treatment; however, its genotoxicity can lead to deoxyribonucleic acid (DNA) damage in healthy cells. Lycopene and Coenzyme Q10 are natural antioxidants capable of exerting protective effects against oxidative damage. This study was conducted to determine the protective effect of lycopene combined with Q10 against 5-FU-induced genotoxicity in colorectal cancer (SW480) and bone marrow mesenchymal stem cells (MSCs) cell lines using the Comet assay.
Methods: This descriptive-analytical study was conducted on SW480 and MSCs cell lines obtained from the Iranian Genetic Resources Cell Bank at the Cell Culture Laboratory of the Faculty of Pharmacy in 2023. The SW480 and MSCs cell lines were cultured at a density of 104 and exposed to a single dose of 5-FU (1 µM) along with various concentrations of lycopene and Q10 (0, 10, 20, and 30 µM). For each cell line, seven groups were defined: A control group (without treatment); a 5-FU group at optimum concentration (1 µM); groups receiving Q10 at 10, 20, and 30 µM plus lycopene at 10, 20, and 30 µM, respectively, comined with receiving 5-FU at optimum concentration (1 µM); a group receiving Q10 alone (30 µM); and a group receiving lycopene alone (30 µM). Cytotoxicity was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, and DNA damage was assessed via the Comet assay.
Results: 5-FU caused a significant decrease in cell viability and a significant increase in DNA damage (P<0.05). Lycopene and Q10 alone did not exhibit significant cytotoxicity. The combination of lycopene and Q10 with 5-FU culminated in increased cell viability and decreased DNA damage compared to the group treated with 5-FU alone.
Conclusion: Lycopene and Q10 demonstrated significant protective effects against 5-FU-induced genotoxicity in both SW480 and MSCs cell lines.
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