Analysis of the protective effect of EGCG against chromosomal damages induced by Nickel treatment of HFF cell line using micronucleus assay on binucleated cells

Document Type : Research Paper


1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

2 Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.



Nickel is a heavy metal, which due to its wide use in industries, found all around our habitat. Nickel-induced chromosomal abnormalities are related to its ability in producing reactive oxygen species (ROS). Use of herbal ingredients in preventing chromosomal abnormalities induced by such elements has been become widely common. Green tea with its polyphenols especially Epigallocatechin gallate (EGCG) which could be used as a common drink is a suitable choice for protecting against heavy metals including Nickel. In this study, effect of three doses of 10, 20 and 30 mM of EGCG on chromosomal abnormalities induced by 0.2, 0.5 and 0.7 mM of Nickel in HFF cell line was investigated using micronucleus assay on Binucleated cells. In different doses, Nickel was able to increase the frequency of Micronucleated Binucleates (MnBi) significantly, which represents chromosomal abnormalities, in comparison to control. While, in all doses mentioned earlier, EGCG was able to dose dependently decrease the frequency of this cells. The results showed that EGCG is able to reduce clastogenic effect of Nickel in vitro. This ability increases in higher doses. Considering the antioxidant ability of EGCG, it could be concluded that Nickel induces most of its chromosomal abnormalities by increasing ROS.


Ali, H., Khan, E., Sajad, M.A., 2013. Phytoremediation of heavy metals--concepts and applications. Chemosphere 91(7), 869-881.
Buxton, S., Voges, Y., Donath, C., Oller, A., 2020. Gene (HPRT) and chromosomal (MN) mutations of nickel metal powder in V79 Chinese hamster cells. Mutation Research 819-820, 111688.
Cameron, K.S., Buchner, V., Tchounwou, P.B., 2011. Exploring the molecular mechanisms of nickel-induced genotoxicity and carcinogenicity: a literature review. Reviews on Environmental Health 26(2), 81-92.
Costa, S., Utan, A., Cervellati, R., Speroni, E., Guerra, M.C., 2007. Catechins: natural free-radical scavengers against ochratoxinA-induced cell damage in a pig kidney cell line (LLC-PK1). Food and Chemical Toxicology 45(10), 1910-1917.
Farhan, M., 2022. Green Tea Catechins: Nature's Way of Preventing and Treating Cancer. International Journal of Molecular Sciences 23(18), 10713.
Fenech, M., 2000. The in vitro micronucleus technique. Mutation Research 455(1-2), 81-95.
Garcia-Rodriguez Mdel, C., Montano-Rodriguez, A.R., Altamirano-Lozano, M.A., 2016. Modulation of hexavalent chromium-induced genotoxic damage in peripheral blood of mice by epigallocatechin-3-gallate (EGCG) and its relationship to the apoptotic activity. Journal of Toxicology and Environmental Health, Part A 79(1), 28-38.
Garcia-Rodriguez Mdel, C., Nicolas-Mendez, T., Montano-Rodriguez, A.R., Altamirano-Lozano, M.A., 2014. Antigenotoxic effects of (-)-epigallocatechin-3-gallate (EGCG), quercetin, and rutin on chromium trioxide-induced micronuclei in the polychromatic erythrocytes of mouse peripheral blood. Journal of Toxicology and Environmental Health, Part A 77(6), 324-336.
Genchi, G., Carocci, A., Lauria, G., Sinicropi, M.S., Catalano, A., 2020. Nickel: Human Health and Environmental Toxicology. International Journal of Environmental Research and Public Health 17(3), 679.
Ghelishli, N., Ghasemi, A., Hosseinimehr, S.J., 2019. The Influence of Piperine on the Radioprotective Effect of Curcumin in Irradiated Human Lymphocytes. Turkish Journal of Pharmaceutical Sciences 16, 366-370.
Ghorbani M, Haddad F, Shahrokhabadi K., 2022. Nickel Increases Chromosomal Abnormalities by Interfering With the Initiation of DNA Repair Pathways. Iranian Journal of Toxicology, 16(3):229-236.
Henkler, F., Brinkmann, J., Luch, A., 2010. The role of oxidative stress in carcinogenesis induced by metals and xenobiotics. Cancers (Basel) 2(2), 376-396.
IARC, 2012. Nickel and nickel compounds. IARC Monograph on the Evaluation of Carcinogenic Risk to Human. 100C, 49.
Kazemi, R., Hosseinimehr, S.J., 2021. Radioprotective Effect of Pioglitazone Against Genotoxicity Induced by Ionizing Radiation in Healthy Human Lymphocytes. Cardiovascular & Hematological Agents in Medicinal Chemistry 19(1), 72-75.
Kopustinskiene, D.M., Jakstas, V., Savickas, A., Bernatoniene, J., 2020. Flavonoids as Anticancer Agents. Nutrients 12(2), 457.
Nas, B., Colak, D.A., 2020. Genotoxic analysis of nickel-iron oxide in Drosophila. Toxicology and Industerial Health 36(11), 835-843.
Ni, J., Guo, X., Wang, H., Zhou, T., Wang, X., 2018. Differences in the Effects of EGCG on Chromosomal Stability and Cell Growth between Normal and Colon Cancer Cells. Molecules 23(4), 788.
Rady, I., Mohamed, H., Rady, M., Siddiqui, I.A., Mukhtar, M., 2018. Cancer preventive and therapeutic effects of EGCG, the major polyphenol in green tea. Egyptian Journal of Basic and Applied Sciences 5(1), 1-23.
Tanaka, K.I., Nakaguchi, S., Shiota, S., Nakada, Y., Oyama, K., Sakakibara, O., Shimoda, M., Sugimoto, A., Ichitani, M., Takihara, T., Kinugasa, H., Kawahara, M., 2022. Preventive Effect of Epigallocatechin Gallate, the Main Component of Green Tea, on Acute Lung Injury Caused by Air Pollutants. Biomolecules 12(9), 1196.
Xie, L.W., Cai, S., Zhao, T.S., Li,M., Tian, Y., 2020. Green tea derivative (-)-epigallocatechin-3-gallate (EGCG) confers protection against ionizing radiation-induced intestinal epithelial cell death both in vitro and in vivo. Free Radical Biology & Medicine 161, 175-186.
Xie, X., Wan, J., Zheng, X., Pan, W., Yuan, J., Hu, B., Feng, M., Liu, Z., Cai, S., 2022. Synergistic effects of epigallocatechin gallate and l-theanine in nerve repair and regeneration by anti-amyloid damage, promoting metabolism, and nourishing nerve cells. Frontiers in Nutrition 9, 951415.
Yi, J., Chen, C., Liu, X., Kang, Q., Hao, L., Huang, J., Lu, J., 2020. Radioprotection of EGCG based on immunoregulatory effect and antioxidant activity against (60)Cogamma radiation-induced injury in mice. Food and Chemical Toxicology 135, 111051.
Zhang, S., Duangjan, C., Tencomnao, T., Wu, L., Wink, M., Lin, J., 2022. Oolonghomobisflavans exert neuroprotective activities in cultured neuronal cells and anti-aging effects in Caenorhabditis elegans. Frontiers in Aging Neuroscience 14, 967316.
Zhao, J., Shi, X., Castranova, V., Ding, M., 2009. Occupational toxicology of nickel and nickel compounds. Journal of Environmental Pathology, Toxicology and Oncology 28(3), 177-208.
Zhong, Y., Ma, C.M., Shahidi, F., 2012. Antioxidant and antiviral activities of lipophilic epigallocatechin gallate (EGCG) derivatives. Journal of Functional Foods 4(1), 87-93.
Zhu, W., Zhao, Y., Zhang, S., Li, X., Xing, L., Zhao, H., Yu, J., 2021. Evaluation of Epigallocatechin-3-Gallate as a Radioprotective Agent During Radiotherapy of Lung Cancer Patients: A 5-Year Survival Analysis of a Phase 2 Study. Frontiers in Oncology 11, 686950.