Journals / Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi / 2026 / Cilt: 16 Sayı: 3

The Role of the Comet Assay in Investigating the Genotoxic and Anti-genotoxic Effects of Plant Secondary Metabolites: A Current Review

Bitki Sekonder Metabolitlerinin Genotoksik ve Anti-genotoksik Etkilerinin Araştırılmasında Komet Testinin Rolü: Güncel Bir Derleme

Merve Yıldız(GAZİ ÜNİVERSİTESİ, FEN FAKÜLTESİ)0009-0007-5023-0532
Deniz Yüzbaşıoğlu(GAZİ ÜNİVERSİTESİ, FEN FAKÜLTESİ)0000-0003-2756-7712
Pages
966–982
Publication Date
2026-09-01
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Due to its sensitivity and reliability, the comet assay is expected to continue to be widely used in future genotoxicity and antigenotoxicity studies.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Plant secondary metabolites (SMs) are organic compounds produced by plants that often exhibit significant pharmacological and biological activities.
📊
Key Findings & Data:Based on the studies reviewed, it has been observed that terpenes mostly have genotoxic effects on various cancer cell lines, whereas phenolic compounds exhibit significant antigenotoxic effects in the tissues studied.
💡
Conclusion & Impact:Due to its sensitivity and reliability, the comet assay is expected to continue to be widely used in future genotoxicity and antigenotoxicity studies.
🔬Design:Review & Systematic Survey
📍Scope:Comprehensive Sample
⚙️Variables:Bitki Sekonder Metabolitleri · Genotoksisite- Anti-genotoksisite · Komet/Tek Hücre Jel Elektroforez Testi

Abstract

Plant secondary metabolites (SMs) are organic compounds produced by plants that often exhibit significant pharmacological and biological activities. They are a primary source for drug discovery and have wide-ranging applications in agriculture, food, textiles, and the dye industry. Despite their beneficial effects, the potential toxicity of specific secondary metabolites can limit their use. Consequently, evaluating the genotoxic and/or antigenotoxic potential of SMs, especially those intended for direct human exposure, is crucial. Various experimental approaches, including in vitro and in vivo methods, have been used for this purpose. This review begins by discussing the definition, classification, and mechanisms of action of plant secondary metabolites. It then focuses on studies that have employed the Comet assay to investigate their genotoxic/antigenotoxic properties. The literature reviewed is limited to studies that utilized the Comet assay, which is widely regarded as a reliable method for detecting DNA strand breaks and assessing genotoxicity/antigenotoxicity. Based on the studies reviewed, it has been observed that terpenes mostly have genotoxic effects on various cancer cell lines, whereas phenolic compounds exhibit significant antigenotoxic effects in the tissues studied. Secondary metabolites containing nitrogen and sulfur, on the other hand, stand out for their antigenotoxic and anticancer properties. In general, the genotoxic, antigenotoxic, anticancer, and antioxidative effects of plant secondary metabolites vary depending on the application concentration, the type of cell or tissue targeted, and the duration of exposure. Further research is needed to elucidate the mechanisms of action of SMs. Due to its sensitivity and reliability, the comet assay is expected to continue to be widely used in future genotoxicity and antigenotoxicity studies.

Özet

Bitki sekonder metabolitleri (SM'ler), genellikle önemli farmakolojik ve biyolojik aktiviteler sergileyen bitkiler tarafından üretilen organik bileşiklerdir. SM’ler, ilaç keşfi için önemli bir kaynaktır ve ayrıca tarım, gıda, tekstil ve boya endüstrisinde geniş kapsamlı uygulamalara sahiptir. Yararlı etkilerine rağmen, bazı sekonder metabolitlerin potansiyel toksisitesi kullanımlarını sınırlayabilmektedir. Sonuç olarak, SM'lerin, özellikle doğrudan insan maruziyeti için tasarlananların, genotoksik ve/veya anti-genotoksik potansiyelinin değerlendirilmesi çok önemlidir. Bu amaçla hem in vitro hem de in vivo yöntemler dahil olmak üzere çeşitli deneysel yaklaşımlar kullanılmaktadır. Bu derleme, bitki sekonder metabolitlerinin tanımı, sınıflandırılması ve etki mekanizmalarını tartışarak başlamaktadır. Daha sonra, genotoksik ve anti-genotoksik özelliklerini araştırmak için Komet testini kullanan çalışmalara odaklanmıştır. İncelenen kaynaklar, DNA ipliğindeki kırıkları tespit etmek ve genotoksisiteyi/anti-genotoksisiteyi değerlendirmek için yaygın olarak uygulanan ve güvenilir bir yöntem olarak kabul edilen Komet testini kullanan çalışmalarla sınırlandırılmıştır. İncelenen çalışmalar doğrultusunda, terpenlerin çoğunlukla çeşitli kanser hücre hatları üzerinde genotoksik etkilere sahip olduğu, buna karşın fenolik bileşiklerin çalışılan dokularda belirgin biçimde anti-genotoksik etkiler gösterdiği görülmektedir. Azot ve sülfür içeren SM’ler ise özellikle anti-genotoksik ve anti-kanser özellikleriyle öne çıkmaktadır. Genel olarak bitki sekonder metabolitlerinin genotoksik, anti-genotoksik, anti-kanser ve anti-oksidatif etkileri; uygulama konsantrasyonuna, hedef alınan hücre veya doku tipine ve maruz kalma süresine bağlı olarak değişkenlik göstermektedir. Bütün bu verilere rağmen, SM’lerin etki mekanizmalarının aydınlatılabilmesi için daha ileri düzeyde araştırmalara ihtiyaç duyulmaktadır. Hassas ve güvenilir bir test olması nedeniyle Komet testinin gelecekteki genotoksisite ve anti-genotoksisite çalışmalarında yaygın olarak kullanılmaya devam etmesi beklenmektedir.

🔖

Cite This Article

Yıldız, M., Yüzbaşıoğlu, D., Ünal, F. (2026). The Role of the Comet Assay in Investigating the Genotoxic and Anti-genotoxic Effects of Plant Secondary Metabolites: A Current Review. Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi, Cilt: 16 Sayı: 3, 966–982. https://doi.org/10.21597/jist.1802516

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (63)

  1. Abdel-Rahman, H. G., Abdelrazek, H. M., Zeidan, D. W., Mohamed, R. M. & Abdelazim, A. M. (2018). Lycopene: hepatoprotective and antioxidant effects toward bisphenol A‐induced toxicity in female Wistar rats. Oxidative Medicine and Cellular Longevity, 2018(1), 5167524.
  2. Akman, Y. ve Düzenli, Sema. (2010). Bitki Fizyolojisi. 186-206 sy.
  3. Anandakumar, P., Kamaraj, S. & Vanitha, M. K. (2021). D‐limonene: A multifunctional compound with potent therapeutic effects. Journal of Food Biochemistry, 45(1), e13566. Arora, D., Bhati, H. & Bansal, K. (2025). Recent advancements in genistein nanocarrier systems for effective cancer management. Medical Oncology, 42(4), 1-23.
  4. Aydın, Ç. ve Mammadov, R. (2017). Insektisit Aktivite Gösteren Bitkisel Sekonder Metabolitlerve Etki Mekanizmasi. Marmara Pharmaceutical Journal, 21(1), 30-37.
  5. Bakkali, F., Averbeck, S., Averbeck, D. & Idaomar, M. (2008). Biological effects of essential oils–a review. Food And Chemical Toxicology, 46(2), 446-475.
  6. Balbaa, M., El-Zeftawy, M. & Abdulmalek, S. A. (2021). Therapeutic Screening of Herbal Remedies for the Management of Diabetes. Molecules, 26(22), Article 6836.
  7. Bi, Y.-l., Min, M., Shen, W. & Liu, Y. (2018). Genistein induced anticancer effects on pancreatic cancer cell lines involve mitochondrial apoptosis, G0/G1cell cycle arrest and regulation of STAT3 signalling pathway. Phytomedicine, 39, 10-16.
  8. Bruić, M., Pirković, A., Vilotić, A., Jovanović-Krivokuća, M. & Spremo-Potparević, B. (2023). Cytoprotective and genoprotective effects of taxifolin against oxidative damage in HTR-8/SVneo human trophoblast cells. Mutagenesis, 38(1), 64-70.
  9. Cardoso, P. C. d. S., Rocha, C. A. M. d., Mota, T. C., Bahia, M. d. O., Correa, R. M. d. S., Gomes, L. M., … & Burbano, R. (2019). In vitro assessment of cytotoxic, genotoxic and mutagenic effects of antimalarial drugs artemisinin and artemether in human lymphocytes. Drug and Chemical Toxicology, 42(6), 608-614.
  10. Cascajosa-Lira, A., Puerto, M., Prieto, A. I., Pichardo, S., Díez-Quijada Jiménez, L., Baños, A.,… & Jos, Á. (2021). Genotoxicity evaluation of Propyl-Propane-Thiosulfinate (PTS) from Allium genus essential oils by a combination of micronucleus and comet assays in rats. Foods, 10(5), 989.
  11. Collins, A., Møller, P., Gajski, G., Vodenková, S., Abdulwahed, A., Anderson, D.,… & Azqueta, A. (2023). Measuring DNA modifications with the comet assay: a compendium of protocols. Nature Protocols, 18(3), 929-989.
  12. Deng, Z., Hassan, S., Rafiq, M., Li, H., He, Y., Cai, Y.,… & Yan, T. (2020). Pharmacological activity of eriodictyol: The major natural polyphenolic flavanone. Evidence‐based Complementary and Alternative Medicine, 2020(1), 6681352.
  13. Dereli Can, G., Taner, G. & İnci Aydemir, Ç. (2023). Evaluation of the cytotoxic and genotoxic/antigenotoxic effects of resveratrol in human limbal explant cultures. International Ophthalmology, 43(6), 1977-1985.
  14. Dutta, S., Wadekar, R. R. & Roy, T. (2021). Radioprotective natural products as alternative complements in oncological radiotherapy. Boletın Latınoamerıcano Y Del Carıbe De Plantas Medıcınales Y Aromátıcas, 20, 101-122.
  15. Erikel, E., Yuzbasioglu, D. & Unal, F. (2020). Genotoxic and antigenotoxic potential of amygdalin on isolated human lymphocytes by the comet assay. Journal of Food Biochemistry, 44(10), e13436.
  16. Fahmy, M. A., Hassan, E. E., Farghaly, A. A. & Hassan, Z. M. (2023). Genotoxicity, DNA damage and sperm defects induced by vinblastine. Molecular Biology Reports, 50(2), 1059-1068.
  17. Fasiku, V., Kyagaba, D., Hlalele, A., Adegoke, A., Erukainure, O. L. & Sekhoacha, M. (2025). A perspective review on factors that influence mutagenicity in medicinal plants and their health implications. Toxicological Sciences.
  18. Gad, H. A., Ramadan, G. R. M., El-Bakry, A. M., El-Sabrout, A. M., Abdelgaleil, S. A. M. Monoterpenes: Promising natural products for public health insect control- A review, International Journal of Tropical Insect Science, 42(2), 1059-1075, 2022.
  19. Glavinić, U., Rajković, M., Ristanić, M., Stevanović, J., Vejnović, B., Djelić, N. & Stanimirović, Z. (2023). Genotoxic Potential of Thymol on Honeybee DNA in the Comet Assay. Insects, 14(5), 451.
  20. Goswami, P., Paul, S., Banerjee, R., Kundu, R. & Mukherjee, A. (2018). Betulinic acid induces DNA damage and apoptosis in SiHa cells. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 828, 1-9.
  21. Hadi, N. S. A., Bankoglu, E. E., Schott, L., Leopoldsberger, E., Ramge, V., Kelber, O.,…& Stopper, H. (2021). Genotoxicity of selected pyrrolizidine alkaloids in human hepatoma cell lines HepG2 and Huh6. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 861, 503305.
  22. Hernandes, L. C., Machado, A. R. T., Tuttis, K., Ribeiro, D. L., Aissa, A. F., Dévoz, P. P. & Antunes, L. M. G. (2020). Caffeic acid and chlorogenic acid cytotoxicity, genotoxicity and impact on global DNA methylation in human leukemic cell lines. Genetics and Molecular Biology, 43(3), e20190347.
  23. Hunyadi, A. (2019). The mechanism (s) of action of antioxidants: From scavenging reactive oxygen/nitrogen species to redox signaling and the generation of bioactive secondary metabolites. Medicinal Research Reviews, 39(6), 2505-2533.
  24. Jamwal, K., Bhattacharya, S. & Puri, S. (2018). Plant growth regulator mediated consequences of secondary metabolites in medicinal plants. Journal of Applied Research on Medicinal and Aromatic Plants, 9, 26-38.
  25. Jin, B., Liu, J., Gao, D., Xu, Y., He, L., Zang, Y.,…& Lin, D. (2020). Detailed studies on the anticancer action of rosmarinic acid in human Hep-G2 liver carcinoma cells: evaluating its effects on cellular apoptosis, caspase activation and suppression of cell migration and invasion. Journal of BUON, 25(3), 1383-1389.
  26. Kaushik, B., Sharma, J., Kumar, P. & Shourie, A. (2021). Phytochemical properties and pharmacological role of plants: secondary metabolites. Biosciences Biotechnology Research Asia, 18(1), 23.
  27. Keskin, B., Orta-Yilmaz, B. & Aydin, Y. (2025). The role of luteolin in modulation of acrylamide-induced genotoxicity and apoptosis in embryonic fibroblast cells. Mutation Research-Genetic Toxicology and Environmental Mutagenesis, 902, 503853.
  28. Koutova, D., Maafi, N., Muthna, D., Kralovec, K., Kroustkova, J., Pidany, F.,… & Rezacova, M. (2023). Antiproliferative activity and apoptosis-inducing mechanism of Amaryllidaceae alkaloid montanine on A549 and MOLT-4 human cancer cells. Biomedicine & Pharmacotherapy, 166, 115295.
  29. Kurt, D., Acar, A., Çavuşoğlu, D., Yalçin, E. & Çavuşoğlu, K. (2021). Genotoxic effects and molecular docking of 1, 4-dioxane: Combined protective effects of trans-resveratrol. Environmental Science and Pollution Research, 28(39), 54922-54935.
  30. Li, X., He, X., Chen, S., Le, Y., Bryant, M. S., Guo, L.,…& Mei, N. (2021). The genotoxicity potential of luteolin is enhanced by CYP1A1 and CYP1A2 in human lymphoblastoid TK6 cells. Toxicology Letters, 344, 58-68.
  31. Liu, X. & Song, L. (2022). Quercetin protects human liver cells from o, p’-DDT-induced toxicity by suppressing Nrf2 and NADPH oxidase-regulated ROS production. Food and Chemical Toxicology, 161, 112849.
  32. Machado, K. L., Marinello, P. C., Silva, T. N. X., Silva, C. F. N., Luiz, R. C., Cecchini, R. & Cecchini, A. L. (2021). Oxidative stress in caffeine action on the proliferation and death of human breast cancer cells MCF-7 and MDA-MB-231. Nutrition and Cancer, 73(8), 1378-1388.
  33. Mahajan, E., Singh, S., Kaur, S., Sohal, S. K. & Sohal, S. K. (2022). The genotoxic, cytotoxic and growth regulatory effects of plant secondary metabolite β-caryophyllene on polyphagous pest Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). Toxicon, 219, Article 106930. https://doi.org/10.1016/j.toxicon.2022.09.016
  34. Majidinia, M., Bishayee, A. & Yousefi, B. (2019). Polyphenols: Major regulators of key components of DNA damage response in cancer. DNA repair, 82, 102679.
  35. Marchese, A., Orhan, I. E., Daglia, M., Barbieri, R., Di Lorenzo, A., Nabavi, S. F.,… & Nabavi, S. M. (2016). Antibacterial and antifungal activities of thymol: A brief review of the literature. Food Chemistry, 210, 402-414.
  36. Nasim, N., Sandeep, I. S. & Mohanty, S. (2022). Plant-derived natural products for drug discovery: current approaches and prospects. Nucleus-India, 65(3), 399-411.
  37. Olive, P. L. & Banáth, J. P. (2006). The comet assay: a method to measure DNA damage in individual cells. Nature Protocols, 1(1), 23-29.
  38. Ostling, O. & Johanson, K. J. (1984). Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochemical and Biophysical Research Communications, 123(1), 291-298.
  39. Parisi, K., Shafee, T. M., Quimbar, P., van der Weerden, N. L., Bleackley, M. R. & Anderson, M. A. (2019). The evolution, function and mechanisms of action for plant defensins. Seminars In Cell & Developmental Biology
  40. Patil, D., Thakur, S., Cota, D. & Paranjape, R. (2023). Evaluation of in-vitro Effect of Polyphenols to prevent DNA Damage in Colon Cell Line after Bacterial Infection. https://doi.org/10.22214/ijraset.2023.56182
  41. Planelló, R., Aquilino, M., Beaugeard, L., Llorente, L., Herrero, Ó., Siaussat, D. & Lécureuil, C. (2024). Unveiling Molecular Effects of the Secondary Metabolite 2-Dodecanone in the Model Hymenopteran Nasonia vitripennis. Toxics, 12(2), 159.
  42. Qi, F., Yan, Q., Zheng, Z., Liu, J., Chen, Y. & Zhang, G. (2018). Geraniol and geranyl acetate induce potent anticancer effects in colon cancer Colo-205 cells by inducing apoptosis, DNA damage and cell cycle arrest. Journal of BUON, 23(2), 346-352.
  43. Raguraman, S., Almoallim, H. S., Alahmadi, T. A., Hussein-Al-Ali, S. H., Bharathi, M. & Ramu, A. K. (2024). Reserpine Induces Apoptosis in Drug-Resistant Cancer through Modulating STAT3 and NF-κB Signaling. Indian Journal of Pharmaceutical Education and Research, 58(3s), s900-s909.
  44. Salah, A., Bouaziz, C., Amara, I., Abid-Essefi, S. & Bacha, H. (2019). Eugenol protects against citrinin-induced cytotoxicity and oxidative damages in cultured human colorectal HCT116 cells. Environmental Science and Pollution Research, 26, 31374-31383.
  45. Seca, A. M. & Pinto, D. C. (2018). Plant secondary metabolites as anticancer agents: successes in clinical trials and therapeutic application. International Journal of Molecular Sciences, 19(1), 263.
  46. Singh, N. P., McCoy, M. T., Tice, R. R. & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175(1), 184-191.
  47. Singh, S., Diksha, Mahajan, E. & Sohal, S. K. (2022). Appraisal of growth inhibitory, biochemical and genotoxic effects of Allyl Isothiocyanate on different developmental stages of Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae). Scientific Reports, 12(1), 10363.
  48. Souza, A. C. S., Silva, L. K., Queiroz, T. B., Marques, E. S., Hiruma-Lima, C. A., Gaivao, I. O. M. & Maistro, E. L. (2020). Citral presents cytotoxic and genotoxic effects in human cultured cells. Drug and Chemical Toxicology, 43(4), 435-440.
  49. Sukhikh, S., Babich, O., Prosekov, A., Kalashnikova, O., Noskova, S., Bakhtiyarova, A.,…& Ivanova, S. (2023). Antidiabetic Properties of Plant Secondary Metabolites. Metabolites, 13(4), 513. https://doi.org/10.3390/metabo13040513
  50. Tasneem, S. & Yasmeen, R. (2018). Evaluation of genotoxicity by comet assay (single-cell gel electrophoresis) in tissues of the fish Cyprinus carpio during sub-lethal exposure to Karanjin. The Journal of Basic and Applied Zoology, 79, 1-13.
  51. The Organisation for Economic Co-operation and Development (OECD), (2016). Test No. 489: In Vivo Mammalian Alkaline Comet Assay. https://www.oecd.org/en/publications/2016/07/test-no-489-in-vivo-mammalian-alkaline-comet-assay_g1g6fb2c.html
  52. Tian, Q., Wang, L., Sun, X., Zeng, F., Pan, Q. & Xue, M. (2019). Scopoletin exerts anticancer effects on human cervical cancer cell lines by triggering apoptosis, cell cycle arrest, inhibition of cell invasion and PI3K/AKT signalling pathway. J BUON, 24(3), 997-1002. Twaij, B. M. & Hasan, M. N. (2022). Bioactive secondary metabolites from plant sources: types, synthesis, and their therapeutic uses. International Journal of Plant Biology, 13(1), 4-14.
  53. Ünal, F., Saygılı Y. ve Akbaş, E. (2022). Kuyruklu Yıldız/Komet Testi. Ed. Prof. Dr. Fatma Ünal, Prof. Dr. Deniz Yüzbaşıoğlu, Genetik Toksikoloji. Nobel Yayıncılık.
  54. Üremiş, N., Üremiş, M. M., Çiğremiş, Y., Tosun, E., Baysar, A. & Türköz, Y. (2022). Cucurbitacin I exhibits anticancer efficacy through induction of apoptosis and modulation of JAK/STAT3, MAPK/ERK, and AKT/mTOR signaling pathways in HepG2 cell line. Journal of Food Biochemistry, 46(10), e14333.
  55. Verma, N., Bal, S., Gupta, R., Aggarwal, N. & Yadav, A. (2020). Antioxidative effects of piperine against cadmium-induced oxidative stress in cultured human peripheral blood lymphocytes. Journal of Dietary Supplements, 17(1), 41-52.
  56. Verma, N., Yadav, A., Bal, S., Gupta, R. & Aggarwal, N. (2019). In vitro studies on ameliorative effects of limonene on cadmium-induced genotoxicity in cultured human peripheral blood lymphocytes. Applied Biochemistry and Biotechnology, 187, 1384-1397.
  57. Verschaeve, L. (2015). Genotoxicity and antigenotoxicity studies of traditional medicinal plants: how informative and accurate are the results? Natural Product Communications, 10(8), 1489-1493.
  58. Vrhovac Madunić, I., Madunić, J., Antunović, M., Paradžik, M., Garaj-Vrhovac, V., Breljak, D.,…& Gajski, G. (2018). Apigenin, a dietary flavonoid, induces apoptosis, DNA damage, and oxidative stress in human breast cancer MCF-7 and MDA MB-231 cells. Naunyn-Schmiedeberg's Archives of Pharmacology, 391, 537-550.
  59. Wallace, R. J. (2004). Antimicrobial properties of plant secondary metabolites. Proceedings of The Nutrition Society, 63(4), 621-629.
  60. Yüzbaşıoğlu, D., Zengin, N. ve Ünal, F. (2014). Gıda koruyucuları ve genotoksisite testleri. Gıda, 39(3), 179-186.
  61. Zajičková, T., Horváthová, E., Kyzek, S., Šályová, E., Túryová, E., Ševčovičová, A. & Gálová, E. (2022). Comparison of cytotoxic, genotoxic, and DNA-protective effects of skyrin on cancerous vs. non-cancerous human cells. International Journal of Molecular Sciences, 23(10), 5339.
  62. Zhang, Y., Zhang, R. & Ni, H. (2020). Eriodictyol exerts potent anticancer activity against A549 human lung cancer cell line by inducing mitochondrial-mediated apoptosis, G2/M cell cycle arrest and inhibition of m-TOR/PI3K/Akt signalling pathway. Archives of Medical Science, 16(2), 446-452.
  63. Zhao, L., Zhang, J., Fan, Y. & Li, Y. (2019). Antiproliferative activity of carnosic acid is mediated via inhibition of cell migration and invasion, and suppression of phosphatidylinositol 3-kinases (PI3K)/AKT/mammalian target of rapamycin (mTOR) signaling pathway. Medical Science Monitor: International Medical Journal of Experimental and Clinical Research, 25, 7864.