Genotoxicity of a synthetic plant growth regulator, Forchlorfenuron (CPPU), on human lymphocytes using chromosome aberration assay
DOI:
https://doi.org/10.36253/caryologia-2006Keywords:
Forchlorfenuron, CPPU, synthetic plant growth regulator, chromosome aberration, mitotic index, human lymphocytesAbstract
Forchlorfenuron (FCF, also known as CPPU), which belongs to the group of phenylurea cytokinins, is one of the most widely used synthetic plant growth regulators (PGRs) worldwide. Although FCF plays a crucial role in cellular growth and differentiation by promoting cell division in plants, it disrupts higher-order septin assembly in other eukaryotic organisms, including humans. Despite its widespread use, no study has been found investigating the genotoxic effects of this synthetic PGR on humans. Hence, this investigation was designed to examine the potential cyto-genotoxicity of a commercial formulation of FCF on human peripheral blood lymphocytes (PBLs) using chromosome aberrations (CAs) and mitotic index (MI) endpoints. The whole blood cultures were treated with 0.25, 0.50, 1.00, and 2.00 µg/ml concentrations of a commercial form of FCF. According to the results, FCF significantly enhanced the percentage of cells containing structural CAs at the concentrations of 1.00 and 2.00 µg/ml for both treatment times (24 and 48 h), in comparison to the negative control (P<0.05). Besides, in cultures exposed to FCF concentrations of 0.50, 1.00, and 2.00 µg/ml, the total CA/cell ratio was significantly higher (P<0.05). In addition, FCF was found to have cytotoxic activity on human PBLs at all treatments (except for the lowest concentration at 24 h) by significantly reducing the MI compared to the negative control (P<0.05). The findings of this investigation indicate the first time that a commercial formulation of FCF (0.50-2.00 µg/ml) may have genotoxic and cytotoxic potential on human lymphocytes.
Downloads
References
Angelis D, Karasmanis EP, Bai X, Spiliotis ET. 2014. In silico docking of forchlorfenuron (FCF) to septins suggests that FCF interferes with GTP binding. PLoS One. vol. 9(5):e96390. https://doi.org/10.1371/journal.pone.0096390
Ayabakt? S, Kocaman, AY. 2020. Cytogenotoxic effects of venlafaxine hydrochloride on cultured human peripheral blood lymphocytes. Drug Chem Toxicol. 43(2):192–199. https://doi.org/10.1080/01480545.2018.1486410
Barral Y, Parra M, Bidlingmaier, S, Snyder M. 1999. Nim1-related kinases coordinate cell cycle progression with the organization of the peripheral cytoskeleton in yeast. Genes Dev. 13(2):176–187. https://doi.org/10.1101/gad.13.2.176
Blum W, Henzi T, Pecze L, Diep KL, Bochet CG, Schwaller B. 2019. The phytohormone forchlorfenuron decreases viability and proliferation of malignant mesothelioma cells in vitro and in vivo. Oncotarget. 10(65):6944–6956. https://doi.org/10.18632/oncotarget.27341
Bonassi S, Norppa H, Ceppi M, Strömberg U, Vermeulen R, Znaor A, Cebulska-Wasilewska A, Fabianova E, Fucic A, Gundy S, Hansteen IL, Knudsen LE, Lazutka J, Rossner P, Sram RJ, Boffetta P. 2008. Chromosomal aberration frequency in lymphocytes predicts the risk of cancer: Results from a pooled cohort study of 22 358 subjects in 11 countries. Carcinogenesis. 29(6):1178–1183. https://doi.org/10.1093/carcin/bgn075
Briggs D. 2003. Environmental pollution and the global burden of disease, Br Med Bull. 68:1–24. https://doi.org/10.1093/bmb/ldg019
Bu Q, Wang X, Xie H, Zhong K, Wu Y, Zhang J, Wang Z, Gao H, Huang Y. 2019. 180 Day Repeated-Dose Toxicity Study on Forchlorfenuron in Sprague–Dawley Rats and Its Effects on the Production of Steroid Hormones. J Agric Food Chem. 67(36):10207–10213. https://doi.org/10.1021/acs.jafc.9b03855
EFSA (European Food Safety Authority). 2017. Conclusion on the peer review of the pesticide risk assessment of the active substance forchlorfenuron. EFSA Journal. 15:1–18. https://doi.org/10.2903/j.efsa.2017.4874
Evans HJ. 1984. Human peripheral blood lymphocytes for the analysis of chromosome aberrations in mutagen tests. In: Kilbey BJ, Legato M, Nichols W, Ramel C. Eds. Handbook of Mutagenicity Test Procedures. Amsterdam: Elsevier Science. p. 405–427.
Enserink JM, Smolka MB, Zhou H, Kolodner RD. 2006. Checkpoint proteins control morphogenetic events during DNA replication stress in Saccharomyces cerevisiae. J Cell Biol. 175(5):729–741. https://doi.org/10.1083/jcb.200605080
Gangadhar PL, Sandeep VH, Rajesh VB. 2020. Profile of plant growth regulator poisoning: A two-year study. Journal of the Indian Society of Toxicology. 16(2):19–22. DOI: 10.31736/2020v16i2/19-22
Gong G, Kam H, Tse YC, Giesy JP, Seto SW, Lee SM. 2021. Forchlorfenuron (CPPU) causes disorganization of the cytoskeleton and dysfunction of human umbilical vein endothelial cells, and abnormal vascular development in zebrafish embryos. Environ Pollut. 271: 115791. https://doi.org/10.1016/j.envpol.2020.115791
Heasley LR, Garcia G 3rd, McMurray MA. 2014. Off-target effects of the septin drug forchlorfenuron on nonplant eukaryotes. Eukaryot Cell. 13(11):1411–1420. https://doi.org/10.1128/EC.00191-14
Heasley LR, McMurray MA. 2016. Small molecule perturbations of septins. Methods Cell Biol. 136:311–319. https://doi.org/10.1016/bs.mcb.2016.03.013
Henzi T, Lannes N, Filgueira L. 2021. Septins in Infections: Focus on Viruses. Pathogens. 10(3): 278. https://doi.org/10.3390/pathogens10030278
Hu Q, Nelson WJ, Spiliotis ET. 2008. Forchlorfenuron alters mammalian septin assembly, organization, and dynamics. J Biol Chem. 283(43):29563–29571. https://doi.org/10.1074/jbc.M804962200
Ivanov AI, Le HT, Naydenov NG, Rieder F. 2021. Novel Functions of the Septin Cytoskeleton: Shaping Up Tissue Inflammation and Fibrosis. Am J Pathol. 191(1):40–51. https://doi.org/10.1016/j.ajpath.2020.09.007
Iwase M, Okada S, Oguchi T, Toh-e A. 2004. Forchlorfenuron, a phenylurea cytokinin, disturbs septin organization in Saccharomyces cerevisiae. Genes Genet Syst. 79(4): 199–206. https://doi.org/10.1266/ggs.79.199
Keaton MA, Lew DJ. 2006. Eavesdropping on the cytoskeleton: progress and controversy in the yeast morphogenesis checkpoint. Curr Opin Microbiol. 9(6):540–546. https://doi.org/10.1016/j.mib.2006.10.004
Kim JG, Takami Y, Mizugami T, Beppu K, Fukuda T, Kataoka I. 2006. CPPU applicatio on size and quality of hardy kiwifruit. Scientia Horticulturae. 110(2):219–222. https://doi.org/10.1016/j.scienta.2006.06.017
Kinoshita M. 2006. Diversity of septin scaffolds. Curr Opin Cell Biol. 18(1):54–60. https://doi.org/10.1016/j.ceb.2005.12.005
Kocaman AY, Topakta? M. 2007. In vitro evaluation of the genotoxicity of acetamiprid in human peripheral blood lymphocytes. Environ Mol Mutagen. 48(6):483–490. https://doi.org/10.1002/em.20309
Kocaman AY, Bucak S. 2016. Genotoxic and cytotoxic effects of flumetralin in human peripheral blood lymphocytes in vitro. Toxicol Ind Health. 32(12):1927–1934. https://doi.org/10.1177/0748233715595142
Kocaman AY, Güven B. 2016. In vitro genotoxicity assessment of the synthetic plant growth regulator, 1-naphthaleneacetamide. Cytotechnology. 68(4):947–956. https://doi.org/10.1007/s10616-015-9847-z
Kocaman AY, K?l?ç, E. 2017. Evaluation of the genotoxicity of commercial formulations of ethephon and ethephon+cyclanilide on Allium cepa L. root meristematic cells. Caryologia. 70(3):229–237. https://doi.org/10.1080/00087114.2017.1329960
Kopecný D, Briozzo P, Popelková H, Sebela M, Koncitíková R, Spíchal L, Nisler J, Madzak C, Frébort I, Laloue M, Houba-Hérin N. 2010. Phenyl- and benzylurea cytokinins as competitive inhibitors of cytokinin oxidase/dehydrogenase: a structural study. Biochimie. 92(8):1052–1062. http://dx.doi.org/10.1016/j.biochi.2010.05.006.
Lew DJ. 2003. The morphogenesis checkpoint: how yeast cells watch their figures. Curr Opin Cell Biol. 15(6):648–653. https://doi.org/10.1016/j.ceb.2003.09.001
Lin LM, Huang DW, Guo JJ, Pan HY, Gong ZY. 2012. Study on toxicity test of forchlorfenuron in mice. Sci Technol Food Ind. 33:360–364. 2012??14?360-362,?3?
Lu Y, Song S, Wang R, Liu Z, Meng J, Sweetman AJ, Jenkins A, Ferrier RC, Li H, Luo W, Wang T. 2015. Impacts of soil and water pollution on food safety and health risks in China. Environ Int. 77:5–15. https://doi.org/10.1016/j.envint.2014.12.010
Marcus EA, Tokhtaeva E, Turdikulova S, Capri J, Whitelegge JP, Scott DR, Sachs G, Berditchevski F, Vagin O. 2016. Septin oligomerization regulates persistent expression of ErbB2/HER2 in gastric cancer cells. Biochem J. 473(12):1703–1718. https://doi.org/10.1042/BCJ20160203
Meng X, Zhai Y, Yuan W, Lv Y, Lv Q, Bai H, Niu Z, Xu W, Ma Q. 2020. Ambient ionization coupled with a miniature mass spectrometer for rapid identification of unauthorized adulterants in food. J Food Compos Anal. 85:103333. https://doi.org/10.1016/j.jfca.2019.103333
Mosesso P, Cinelli S, Natarajan AT, Palitti F. 2013. In Vitro Cytogenetic Assays: Chromosomal Aberrations and Micronucleus Tests. In: Dhawan A, Bajpayee M. (eds). Genotoxicity Assessment. Methods in Molecular Biology, Humana Press, Totowa, NJ. 1044, p. 123-146. https://doi.org/10.1007/978-1-62703-529-3_6
Murgia E, Ballardin M, Bonassi S, Rossi AM, Barale R. 2008. Validation of micronuclei frequency in peripheral blood lymphocytes as early cancer risk biomarker in a nested case-control study. Mutat Res. 639(1-2):27–34. https://doi.org/10.1016/j.mrfmmm.2007.10.010
Ozel CA, Unal F, Avuloglu-Yilmaz E, Erikel E, Mirici S, Yuzbasioglu D. 2022. Determination of genotoxic damages of picloram and dicamba with comet assay in Allium cepa rooted in tissue culture and distilled water. Mol Biol Rep. 49(12):11273-11280. https://doi.org/10.1007/s11033-022-07712-7
Özkul M, Özel ÇA, Yüzba??o?lu D, Ünal F. 2016. Does 2,4-dichlorophenoxyacetic acid (2,4-D) induce genotoxic effects in tissue cultured Allium roots? Cytotechnology. 68(6):2395–2405. https://doi.org/10.1007/s10616-016-9956-3
Phillips DH, Arlt VM. 2009. Genotoxicity: damage to DNA and its consequences. In: Luch A, Ed. Molecular, Clinical and Environmental Toxicology. Experientia Supplementum, 99, p. 87–110. Birkhäuser Basel. https://doi.org/10.1007/978-3-7643-8336-7_4
Rodrigues ET, Alpendurada MF, Ramos F, Pardal MÂ. 2018. Environmental and human health risk indicators for agricultural pesticides in estuaries. Ecotoxicol Environ Saf.150:224–231. https://doi.org/10.1016/j.ecoenv.2017.12.047
Sidhaye VK, Chau E, Breysse PN, King LS. 2011. Septin-2 mediates airway epithelial barrier function in physiologic and pathologic conditions. Am J Respir Cell Mol Biol. 45(1):120–126. https://doi.org/10.1165/rcmb.2010-0235OC
Smolka MB, Chen SH, Maddox PS, Enserink JM, Albuquerque CP, Wei XX, Desai A, Kolodner RD, Zhou H. 2006. An FHA domain-mediated protein interaction network of Rad53 reveals its role in polarized cell growth. J Cell Biol. 175(5):743–753. https://doi.org/10.1083/jcb.200605081
Sharma S, Quintana A, Findlay GM, Mettlen M, Baust B, Jain M, Nilsson R, Rao A, Hogan PG. 2013. An siRNA screen for NFAT activation identifies septins as coordinators of store-operated Ca2+ entry. Nature. 499(7457):238–242. https://doi.org/10.1038/nature12229
Shi X, Jin F, Huang Y, Du X, Li C, Wang M, Shao H, Jin M, Wang J. 2012. Simultaneous determination of five plant growth regulators in fruits by modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction and liquid chromatography-tandem mass spectrometry. J Agric Food Chem. 60(1):60–65. https://doi.org/10.1021/jf204183d
Shulewitz MJ, Inouye CJ, Thorner J. 1999. Hsl7 localizes to a septin ring and serves as an adapter in a regulatory pathway that relieves tyrosine phosphorylation of Cdc28 protein kinase in Saccharomyces cerevisiae. Mol Cell Biol. 19(10):7123–7137. https://doi.org/10.1128/MCB.19.10.7123
Su L, Wang M, Wang Y, Sharif R, Ren N, Qian C, Xu J, Chen X, Qi X. 2021. Forchlorfenuron Application Induced Parthenocarpic Fruit Formation without Affecting Fruit Quality of Cucumber. Horticulturae. 7(6):128. https://doi.org/10.3390/horticulturae7060128
Sun L, Cao X, Lechuga S, Feygin A, Naydenov NG, Ivanov AI. 2020. A Septin Cytoskeleton-Targeting Small Molecule, Forchlorfenuron, Inhibits Epithelial Migration via Septin-Independent Perturbation of Cellular Signaling. Cells. 9(1):84. https://doi.org/10.3390/cells9010084
Tixier C, Sancelme M, Bonnemoy F, Cuer A, Veschambre H. 2001. Degradation products of a phenylurea herbicide, diuron: synthesis, ecotoxicity, and biotransformation. Environ Toxicol Chem. 20(7):1381–1389. https://doi.org/10.1002/etc.5620200701
Tokhtaeva E, Capri J, Marcus EA, Whitelegge JP, Khuzakhmetova V, Bukharaeva E, Deiss-Yehiely N, Dada LA, Sachs G, Fernandez-Salas E, Vagin O. 2015. Septin dynamics are essential for exocytosis. J Biol Chem. 290(9):5280–5297. https://doi.org/10.1074/jbc.M114.616201
Toumi K, Joly L, Vleminckx C, Schiffers B. 2018. Assessment of Belgian florists exposure to pesticide residues. Comm Appl Biol Sci. Ghent University 83:355–368.
USEPA (U.S. Environmental Protection Agency). 2004. Pesticide Fact Sheet for new chemical: Forchlorfenuron; issued: September 2004. United States Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances (7501C). https://www3.epa.gov/pesticides/chem_search/reg_actions/registration/fs_PC-128819_01-Sep-04.pdf
Vardi-Oknin D, Golan M, Mabjeesh NJ. 2013. Forchlorfenuron disrupts SEPT9_i1 filaments and inhibits HIF-1. PLoS One. 8(8):e73179. https://doi.org/10.1371/journal.pone.0073179
Wang KS, Lu CY, Chang SH. 2011. Evaluation of acute toxicity and teratogenic effects of plant growth regulators by Daphnia magna embryo assay. J Hazard Mater. 190(1-3):520–528. https://doi.org/10.1016/j.jhazmat.2011.03.068
Wang Q, Yang Z. 2016.Industrial water pollution, water environment treatment, and health risks in China. Environ Pollut. 218:358-365. https://doi.org/10.1016/j.envpol.2016.07.011
Wang M, Yan H, Han Y, Qiao F. 2023. Miniaturized extraction and determination of swelling agents in fruits and vegetables based on deep eutectic solvent-molecularly imprinted hydrophilic resin. Food Chem. 398:133920. https://doi.org/10.1016/j.foodchem.2022.133920
Weirich CS, Erzberger JP, Barral Y. 2008. The septin family of GTPases: architecture and dynamics. Nat Rev Mol Cell Biol. 9(6):478–489. https://doi.org/10.1038/nrm2407
Zhang N, Liu L, Fan N, Zhang Q, Wang W, Zheng M, Ma L, Li Y, Shi L. 2016. The requirement of SEPT2 and SEPT7 for migration and invasion in human breast cancer via MEK/ERK activation. Oncotarget. 7(38):61587–61600. https://doi.org/10.18632/oncotarget.11402
Zhang W, Jiang F, Ou J. 2011. Global pesticide consumption and pollution: with China as a focus. Proc Int Acad Ecol Environ Sci. 1(2):125–144.
Zhang Z, Guo K, Bai Y, Dong J, Gao Z, Yuan Y, Wang Y, Liu L, Yue T. 2015. Identification, synthesis, and safety assessment of forchlorfenuron (1-(2-chloro-4-pyridyl)-3-phenylurea) and its metabolites in kiwifruits. J Agric Food Chem. 63(11):3059–3066. https://doi.org/10.1021/acs.jafc.5b01100
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 AY?E YAVUZ KOCAMAN
This work is licensed under a Creative Commons Attribution 4.0 International License.
- Copyright on any open access article in a journal published byCaryologia is retained by the author(s).
- Authors grant Caryologia a license to publish the article and identify itself as the original publisher.
- Authors also grant any third party the right to use the article freely as long as its integrity is maintained and its original authors, citation details and publisher are identified.
- The Creative Commons Attribution License 4.0 formalizes these and other terms and conditions of publishing articles.
- In accordance with our Open Data policy, the Creative Commons CC0 1.0 Public Domain Dedication waiver applies to all published data in Caryologia open access articles.
