Молекулярные эффекты пестицидов карбарила, хлорпирифоса, манкоцеба, тирама и пендиметалина в условно-нормальных клетках in vitro: генотоксичность, клоногенность и влияние на экспрессию генов, ассоциированных с канцерогенезом

Введение. Злокачественные новообразования остаются одной из основных причин смертности в мире. В развитии данной патологии большую роль играет воздействие неблагоприятных факторов окружающей среды, в том числе пестицидов. Несмотря на широкий спектр используемых в сельском хозяйстве пестицидов, их молекулярные эффекты и канцерогенный потенциал изучены лишь в отношении небольшого числа моделей, включая нормальные клетки человека.
Цель исследования – изучить молекулярные эффекты пестицидов карбарила, хлорпирифоса, манкоцеба, тирама и пендиметалина в условно-нормальных клетках HaCaT и MCF10A.
Материалы и методы. Нетоксичные концентрации пестицидов определяли с помощью МТТ-теста. Генотоксичность анализировали методом ДНК-комет. Пролиферативный потенциал оценивали с помощью клоногенного анализа, изменение экспрессии генов, ассоциированных с канцерогенезом, – с использованием полимеразной цепной реакции в реальном времени.
Результаты. Карбарил вызывал повреждение ДНК в клетках MCF10A, способствовал пролиферации клеток обеих линий в клоногенном тесте, а также приводил к активации генов биотрансформации (AHR, GSTA4) в клетках MCF10A, репрессии (CYP1B1, GSTA4) в клетках HaCaT и снижению экспрессии генов воспаления (IL1a, IL1b, PTGES, IFNGR1). Хлорпирифос не показал генотоксического эффекта и не влиял на клоногенность, но вызывал индукцию генов биотрансформации (CYP1A1, CYP1B1), воспаления (IL1b, PTGES) и генов BCL2 и DNMTs. Манкоцеб и тирам не проявляли генотоксичности в клетках HaCaT и MCF10A, но активировали отдельные гены репарации (ATR/ATM). Тирам стимулировал пролиферацию клеток HaCaT в клоногенном тесте, а манкоцеб активировал экспрессию генов – регуляторов пролиферации (CCND2, CCNE1, Ki-67), но не влиял на рост колоний; оба фунгицида снижали экспрессию генов воспаления (COX2, IL1a, IL1b). Пендиметалин вызывал повреждение ДНК и активацию экспрессии генов репарации (ATR, GADD45a, PCNA) в клетках обеих линий, а также снижал экспрессию GLUT3 в клетках HaCaT и индуцировал экспрессию генов CYP1A1 в клетках HaCaT и CYP1B1 – в клетках MCF10A.
Заключение. В ходе комплексной оценки влияния пестицидов на нормальные человеческие клетки выявлено, что пендиметалин, хлорпирифос и карбарил оказывают наибольшее проканцерогенное действие.

1. World Health Organization. Global cancer burden growing, amidst mounting need for services. Available at: https://www.who.int/ru/news/item/01-02-2024-global-cancer-burden-growing--amidstmounting-need-for-services

2. Cancer Tomorrow – IARC. Estimated number of new cases from 2022 to 2050, Both sexes, age [0–85+]. Available at: https://gco.iarc.fr/tomorrow/en/dataviz/isotype?years=2050

3. Wan N.F., Fu L., Dainese M. et al. Pesticides have negative effects on non-target organisms. Nat Commun 2025;16(1):1360. DOI: 10.1038/s41467-025-56732-x

4. Stockholm Convention. The 12 initial POPs under the Stockholm Convention. Available at: https://chm.pops.int/Convention/ThePOPs/The12initialPOPs/tabid/296

5. IARC. Agents classified by the IARC Monographs, Volumes 1–139. 2025. Available at: https://monographs.iarc.who.int/agentsclassified-by-the-iarc/

6. Food and Agriculture Organisation of the United Nations. Commission on Genetic Resources for Food and Agriculture. 2019. Available at: https://openknowledge.fao.org/server/api/core/bitstreams/50b79369-9249-4486-ac07-9098d07df60a/content

7. IARC. Report of the Advisory Group to Recommend Priorities for the IARC Monographs during 2025–2029. 2024. Available at: https://www.iarc.who.int/news-events/advisory-grouprecommendations-on-priorities-for-the-iarc-monographsduring-2025-2029/

8. Smith M.T., Guyton K.Z., Gibbons C.F. et al. Key Characteristics of carcinogens as a basis for organizing data on mechanisms of carcinogenesis. Environ Health Perspect 2016;124(6):713–21. DOI: 10.1289/ehp.1509912

9. Ataei M., Abdollahi M. A systematic review of mechanistic studies on the relationship between pesticide exposure and cancer induction. Toxicol Appl Pharmacol 2022;456:116280. DOI: 10.1016/j.taap.2022.116280

10. Shekhar C., Khosya R., Thakur K. et al. A systematic review of pesticide exposure, associated risks, and long-term human health impacts. Toxicol Rep 2024;13:101840. DOI: 10.1016/j.toxrep.2024.101840

11. Sule R.O., Condon L., Gomes A.V. A Common Feature of Pesticides: Oxidative Stress-The Role of Oxidative Stress in Pesticide-Induced Toxicity. Oxid Med Cell Longev. 2022;2022:5563759. DOI: 10.1155/2022/5563759

12. Gunasekara A.S., Rubin A.L., Goh K.S. et al. Environmental fate and toxicology of carbaryl. Rev Environ Contam Toxicol. 2008;196:95-121. DOI: 10.1007/978-0-387-78444-1_4

13. De Roos A.J., Schinasi L.H., Miligi L. et al. Occupational insecticide exposure and risk of non-Hodgkin lymphoma: A pooled case-control study from the InterLymph Consortium. Int J Cancer 2021;149(10):1768-86. DOI: 10.1002/ijc.33740

14. Presutti R., Harris S.A., Kachuri L. et al. Pesticide exposures and the risk of multiple myeloma in men: An analysis of the North American Pooled Project. Int J Cancer 2016;139(8):1703–14. DOI: 10.1002/ijc.30218

15. Koutros S., Harris S.A., Spinelli J.J. et al. Non-Hodgkin lymphoma risk and organophosphate and carbamate insecticide use in the north American pooled project. Environ Int 2019;127:199–205. DOI: 10.1016/j.envint.2019.03.018

16. Erickson P.A., Andreotti G., Remigio R.V. et al. Carbaryl use and incident cancer in the Agricultural Health Study: an updated analysis. Int J Hyg Environ Health 2025;268:114615. DOI: 10.1016/j.ijheh.2025.114615

17. Shukla Y., Antony M., Mehrotra N.K. Carcinogenic and cocarcinogenic studies with carbaryl following topical exposure in mice. Cancer Lett 1992;62(2):133–40. DOI: 10.1016/0304-3835(92)90183-v

18. Meeker J.D., Singh N.P., Ryan L. et al. Urinary levels of insecticide metabolites and DNA damage in human sperm. Hum Reprod 2004;19(11):2573–80. DOI: 10.1093/humrep/deh444

19. Nandi N.K., Vyas A., Akhtar M.J., Kumar B. The growing concern of chlorpyrifos exposures on human and environmental health. Pestic Biochem Physiol 2022;185:105138. DOI: 10.1016/j.pestbp.2022.105138

20. Lee W.J., Blair A., Hoppin J.A. et al. Cancer incidence among pesticide applicators exposed to chlorpyrifos in the Agricultural Health Study. J Natl Cancer Inst 2004;96(23):1781–9. DOI: 10.1093/jnci/djh324

21. Tayour C., Ritz B., Langholz B. et al. A case-control study of breast cancer risk and ambient exposure to pesticides. Environ Epidemiol 2019;3(5):e070. DOI: 10.1097/EE9.0000000000000070

22. Ventura C., Nieto M.R., Bourguignon N. et al. Pesticide chlorpyrifos acts as an endocrine disruptor in adult rats causing changes in mammary gland and hormonal balance. J Steroid Biochem Mol Biol 2016;156:1–9. DOI: 10.1016/j.jsbmb.2015.10.010

23. Hazarika J., Ganguly M., Borgohain G. et al. Endocrine disruption: molecular interactions of chlorpyrifos and its degradation products with estrogen receptor. Structural Chemistry 2020;31:2011–21. DOI: 10.1007/s11224-020-01562-4

24. Leon M.E., Schinasi L.H., Lebailly P. et al. Pesticide use and risk of non-Hodgkin lymphoid malignancies in agricultural cohorts from France, Norway and the USA: a pooled analysis from the AGRICOH consortium. Int J Epidemiol 2019;48(5):1519–35. DOI: 10.1093/ije/dyz017

25. Koutros S., Silverman D.T., Alavanja M.C. et al. Occupational exposure to pesticides and bladder cancer risk. Int J Epidemiol 2016;45(3):792–805. DOI: 10.1093/ije/dyv195

26. Dennis L.K., Lynch C.F., Sandler D.P., Alavanja M.C. Pesticide use and cutaneous melanoma in pesticide applicators in the agricultural heath study. Environ Health Perspect 2010;118(6):812–7. DOI: 10.1289/ehp.0901518

27. Piel C., Pouchieu C., Carles C. et al. Agricultural exposures to carbamate herbicides and fungicides and central nervous system tumour incidence in the cohort AGRICAN. Environ Int 2019;130:104876. DOI: 10.1016/j.envint.2019.05.070

28. Mills P.K., Yang R., Riordan D. Lymphohematopoietic cancers in the United Farm Workers of America (UFW), 1988–2001. Cancer Causes Control 2005;16(7):823–30. DOI: 10.1007/s10552-005-2703-2

29. Vighi M., Matthies M., Solomon K.R. Critical assessment of pendimethalin in terms of persistence, bioaccumulation, toxicity, and potential for long-range transport. J Toxicol Environ Health B Crit Rev 2017;20(1):1–21. DOI: 10.1080/10937404.2016.1222320

30. Hou L., Lee W.J., Rusiecki J. et al. Pendimethalin exposure and cancer incidence among pesticide applicators. Epidemiology 2006;17(3):302–7. DOI: 10.1097/01.ede.0000201398.82658.50

31. Andreotti G., Freeman L.E., Hou L. et al. Agricultural pesticide use and pancreatic cancer risk in the Agricultural Health Study Cohort. Int J Cancer 2009;124(10):2495–500. DOI: 10.1002/ijc.24185

32. Walsh K.D., Kato T.A. Alkaline comet assay to detect DNA damage. Methods Mol Biol 2023;2519:65–72. DOI: 10.1007/978-1-0716-2433-3_7

33. Guzman C., Bagga M., Kaur A. et al. ColonyArea: an ImageJ plugin to automatically quantify colony formation in clonogenic assays. PLoS One 2014;9(3):e92444. DOI: 10.1371/journal.pone.0092444

34. Livak K.J., Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001;25(4):402–8. DOI: 10.1006/meth.2001.1262

35. IARC. Certain polycyclic aromatic hydrocarbons and heterocyclic compounds. 1973. Available at: https://publications.iarc.who.int/Book-And-Report-Series/Iarc-Monographs-On-TheIdentification-Of-Carcinogenic-Hazards-To-Humans/CertainPolycyclic-Aromatic-Hydrocarbons-And-HeterocyclicCompounds-1973

36. Robles H. Methylcholanthrene, 3-. Available at: https://www.researchgate.net/publication/304034151_Methylcholanthrene_3-

37. Kawabe M., Urano K., Suguro M. et al. Tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in an ultra-short-term skin carcinogenesis bioassay using rasH2 mice. Vet Pathol 2013;50(5):903–8. DOI: 10.1177/0300985813486811

38. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature 1984;308(5961):693–8. DOI: 10.1038/308693a0

39. Denison M.S., Phelan D., Winter G.M., Ziccardi M.H. Carbaryl, a carbamate insecticide, is a ligand for the hepatic Ah (dioxin) receptor. Toxicol Appl Pharmacol 1998;152(2):406–14. DOI: 10.1006/taap.1998.9999

40. Larigot L., Juricek L., Dairou J., Coumoul X. AhR signaling pathways and regulatory functions. Biochim Open 2018;7:1–9. DOI: 10.1016/j.biopen.2018.05.001

41. Ledirac N., Delescluse C., de Sousa G. et al. Carbaryl induces CYP1A1 gene expression in HepG2 and HaCaT cells but is not a ligand of the human hepatic Ah receptor. Toxicol Appl Pharmacol 1997;144(1):177–82. DOI: 10.1006/taap.1997.8120

42. Honkakoski P., Negishi M. Regulation of cytochrome P450 (CYP) genes by nuclear receptors. Biochem J 2000;347(Pt 2):321–37. DOI: 10.1042/0264-6021:3470321

43. Saquib Q., Siddiqui M.A., Ansari S.M. et al. Cytotoxicity and genotoxicity of methomyl, carbaryl, metalaxyl, and pendimethalin in human umbilical vein endothelial cells. J Appl Toxicol 2021;41(5):832–46. DOI: 10.1002/jat.4139

44. Fernandes R., Hosoya K., Pereira P. Reactive oxygen species downregulate glucose transport system in retinal endothelial cells. Am J Physiol Cell Physiol 2011;300(4):C927–36. DOI: 10.1152/ajpcell.00140.2010

45. Chen Y., Joo J., Chu J.M. et al. Downregulation of the glucose transporter GLUT 1 in the cerebral microvasculature contributes to postoperative neurocognitive disorders in aged mice. J Neuroinflammation 2023;20(1):237. DOI: 10.1186/s12974-023-02905-8

46. Jorsaraei S.G., Maliji G., Azadmehr A. et al. Immunotoxicity effects of carbaryl in vivo and in vitro. Environ Toxicol Pharmacol 2014;38(3):838–44. DOI: 10.1016/j.etap.2014.09.004

47. Seher Karsli S.Y., Esra F. İncedere düzdağ, Türkan yurdun. Assessment of genotoxic effects of organophosphate and carbamate pesticides by comet assay. İstanbul J Pharm 2022;52(2):136–42. DOI: 10.26650/IstanbulJPharm.2022.1057224

48. Thakur S., Dhiman M., Mantha A.K. APE1 modulates cellular responses to organophosphate pesticide-induced oxidative damage in non-small cell lung carcinoma A549 cells. Mol Cell Biochem 2018;441(1–2):201–16. DOI: 10.1007/s11010-017-3186-7

49. Balakrishnan P., Thirunavukarasu K., Tamizhmani P. Toxicological impact of chronic chlorpyrifos exposure: DNA damage and epigenetic alterations induces neoplastic transformation of liver cells. Biochem Biophys Res Commun 2025;746:151287. DOI: 10.1016/j.bbrc.2025.151287

50. Moyano P., Garcia J., Garcia J.M. et al. Chlorpyrifos-induced cell proliferation in human breast cancer cell lines differentially mediated by estrogen and aryl hydrocarbon receptors and KIAA1363 enzyme after 24 h and 14 days exposure. Chemosphere 2020;251:126426. DOI: 10.1016/j.chemosphere.2020.126426

51. Hevir N., Trost N., Debeljak N., Rizner T.L. Expression of estrogen and progesterone receptors and estrogen metabolizing enzymes in different breast cancer cell lines. Chem Biol Interact 2011; 191(1–3):206–16. DOI: 10.1016/j.cbi.2010.12.013

52. Lasagna M., Ventura C., Hielpos M.S. et al. Endocrine disruptor chlorpyrifos promotes migration, invasion, and stemness phenotype in 3D cultures of breast cancer cells and induces a wide range of pathways involved in cancer progression. Environ Res 2022;204(Pt A):111989. DOI: 10.1016/j.envres.2021.111989

53. Moyano P., Garcia J.M., Garcia J. et al. Chlorpyrifos induces cell proliferation in MCF-7 and MDA-MB-231 cells, through cholinergic and Wnt/beta-catenin signaling disruption, AChE-R upregulation and oxidative stress generation after single and repeated treatment. Food Chem Toxicol 2021;152:112241. DOI: 10.1016/j.fct.2021.112241

54. Ventura C., Nunez M., Miret N. et al. Differential mechanisms of action are involved in chlorpyrifos effects in estrogen-dependent or -independent breast cancer cells exposed to low or high concentrations of the pesticide. Toxicol Lett 2012;213(2):184–93. DOI: 10.1016/j.toxlet.2012.06.017

55. Croom E.L., Wallace A.D., Hodgson E. Human variation in CYPspecific chlorpyrifos metabolism. Toxicology 2010;276(3):184–91. DOI: 10.1016/j.tox.2010.08.005

56. Montanari C., Franco-Campos F., Taroncher M. et al. Chlorpyrifos induces cytotoxicity via oxidative stress and mitochondrial dysfunction in HepG2 cells. Food Chem Toxicol 2024;192:114933. DOI: 10.1016/j.fct.2024.114933

57. Radhey S. Verma A.M., Nalini Srivastava. In vivo chlorpyrifos induced oxidative stress: Attenuation by antioxidant vitamins. Pesticide Biochemistry and Physiology 2007;88(2):191–6. DOI: 10.1016/j.pestbp.2006.11.002

58. Lasagna M., Hielpos M.S., Ventura C. et al. Chlorpyrifos subthreshold exposure induces epithelial-mesenchymal transition in breast cancer cells. Ecotoxicol Environ Saf 2020;205:111312. DOI: 10.1016/j.ecoenv.2020.111312

59. Yahia D., El-Amir Y.O., Rushdi M. Mancozeb fungicide-induced genotoxic effects, metabolic alterations, and histological changes in the colon and liver of Sprague Dawley rats. Toxicol Ind Health 2019;35(4):265–76. DOI: 10.1177/0748233719834150

60. Pienkowska M., Zielenska M. Genotoxic effects of thiram evaluated by sister-chromatid exchanges in human lymphocytes. Mutat Res 1990;245(2):119–23. DOI: 10.1016/0165-7992(90)90010-h

61. Maksimova V., Bukina A., Khayrieva G. et al. Thiram effects on HeLa TI cells. Proceedings 2024;102(1):35. DOI: 10.3390/proceedings2024102035

62. Lori G., Tassinari R., Narciso L. et al. Toxicological Comparison of mancozeb and zoxamide fungicides at environmentally relevant concentrations by an in vitro approach. Int J Environ Res Public Health 2021;18(16):8591. DOI: 10.3390/ijerph18168591

63. Bhaskar R., Mishra A., Mohanty B. Effects of mancozeb and imidacloprid pesticides on activities of steroid biosynthetic enzymes cytochromes P450. J Kalash Sci 2014;2:1–6.

64. Dalvi P.S., Wilder-Ofie T., Mares B. et al. Toxicologic implications of the metabolism of thiram, dimethyldithiocarbamate and carbon disulfide mediated by hepatic cytochrome P450 isozymes in rats. Pesticide Biochemi Physiol 2002;74(2):85–90.

65. Dalvi P.S., Wilder-Ofie T., Mares B. et al. Effect of cytochrome P450 inducers on the metabolism and toxicity of thiram in rats. Vet Hum Toxicol 2002;44(6):331–3.

66. Kumar K., Sabarwal A., Singh R.P. Mancozeb selectively induces mitochondrial-mediated apoptosis in human gastric carcinoma cells through ROS generation. Mitochondrion 2019;48:1–10. DOI: 10.1016/j.mito.2018.06.003

67. Kurpios-Piec D., Grosicka-Maciag E., Wozniak K. et al. Thiram activates NF-kappaB and enhances ICAM-1 expression in human microvascular endothelial HMEC-1 cells. Pestic Biochem Physiol 2015;118:82–9. DOI: 10.1016/j.pestbp.2014.12.003

68. Sarigol Kilic Z., Aydin S., Undeger Bucurgat U., Basaran N. In vitro genotoxicity assessment of dinitroaniline herbicides pendimethalin and trifluralin. Food Chem Toxicol 2018;113:90–8. DOI: 10.1016/j.fct.2018.01.034

69. Ansari S.M., Saquib Q., Attia S.M. et al. Pendimethalin induces oxidative stress, DNA damage, and mitochondrial dysfunction to trigger apoptosis in human lymphocytes and rat bone-marrow cells. Histochem Cell Biol 2018;149(2):127–41. DOI: 10.1007/s00418-017-1622-0

70. Ahmad M.I., Zafeer M.F., Javed M., Ahmad M. Pendimethalininduced oxidative stress, DNA damage and activation of antiinflammatory and apoptotic markers in male rats. Sci Rep 2018;8(1):17139. DOI: 10.1038/s41598-018-35484-3

71. Lee H.S., Amarakoon D., Tamia G. et al. Pendimethalin induces apoptotic cell death through activating ER stress-mediated mitochondrial dysfunction in human umbilical vein endothelial cells. Food Chem Toxicol 2022;168:113370. DOI: 10.1016/j.fct.2022.113370

72. Ham J., Lim W., Song G. Pendimethalin induces apoptosis in testicular cells via hampering ER-mitochondrial function and autophagy. Environ Pollut 2021;278:116835. DOI: 10.1016/j.envpol.2021.116835

73. Sarigol-Kilic Z., Undeger-Bucurgat U. The Apoptotic and Antiapoptotic Effects of Pendimethalin and Trifluralin on A549 Cells In Vitro. Turk J Pharm Sci 2018;15(3):364–9. DOI: 10.4274/tjps.94695