University of Lodz, Faculty of Biology and Environmental Protection, Department of Biophysics of Environmental Pollution, Pomorska Str. 141/143, 90-236 Lodz, Poland; Medical University of Lodz, Department of Internal Diseases and Clinical Pharmacology, Laboratory of Tissue Immunopharmacology, Kniaziewicza Str. 1/5, 91-347 Lodz, Poland.
Nofer Institute of Occupational Medicine, Department of Molecular Genetics and Epigenetics, Teresy Str. 8, 91-348 Lodz, Poland.
Toxicol In Vitro. 2020 Aug;66:104878. doi: 10.1016/j.tiv.2020.104878. Epub 2020 May 1.
Aminomethylphosphonic acid (AMPA) is a primary metabolite of glyphosate and amino-polyphosphonate. We have determined the effect of AMPA on selected epigenetic parameters and major cell cycle drivers in human peripheral blood mononuclear cells (PBMCs). The cells were incubated with AMPA at 0.5, 10 and 250 μM for 24 h. The performed analysis included: global DNA methylation by colorimetric measurement of 5-methylcytosine in DNA, methylation in the promoter regions of selected tumor suppressor genes (P16, P21, TP53) and proto-oncogenes (BCL2, CCND1) as well as the expression profile of the indicated genes by Real-Time PCR assays. The obtained results have revealed significant reduction of global DNA methylation level in PBMCs exposed to AMPA. Investigated xenobiotic changed methylation pattern of the P21 and TP53 suppressor gene promoters, but in case of other analyzed genes: P16, BCL2 and CCND1 no statistically significant changes have been noted. Gene profiling have shown that AMPA only changed the expression of CCND1. Summing up, our results have revealed a small potential disturbance in methylation processes and the absence of changes in expression of tested tumor suppressor genes (P16, P21, TP53) and protooncogenes (BCL2) in human PBMCs exposed to AMPA.
氨甲基膦酸（AMPA）是草甘膦和氨基多膦酸的主要代谢物。我们已经确定了 AMPA 对人外周血单核细胞（PBMCs）中选定的表观遗传参数和主要细胞周期驱动因素的影响。将细胞在 0.5、10 和 250μM 的 AMPA 中孵育 24 小时。进行的分析包括：通过测量 DNA 中 5-甲基胞嘧啶的比色法测定全基因组 DNA 甲基化，选定的肿瘤抑制基因（P16、P21、TP53）和原癌基因（BCL2、CCND1）启动子中的甲基化，以及通过实时 PCR 检测指示基因的表达谱。获得的结果表明，暴露于 AMPA 的 PBMCs 中全基因组 DNA 甲基化水平显着降低。研究的外源性化学物质改变了 P21 和 TP53 抑制剂基因启动子的甲基化模式，但在其他分析的基因（P16、BCL2 和 CCND1）中未观察到统计学上显着的变化。基因谱显示，AMPA 仅改变了 CCND1 的表达。总之，我们的结果表明，暴露于 AMPA 的人 PBMCs 中，甲基化过程中存在微小的潜在干扰，并且测试的肿瘤抑制基因（P16、P21、TP53）和原癌基因（BCL2）的表达没有变化。
