Malinowska Kinga, Tarhonska Kateryna, Foksiński Marek, Sicińska Paulina, Jabłońska Ewa, Reszka Edyta, Zarakowska Ewelina, Gackowski Daniel, Górecka Karolina, Balcerczyk Aneta, Bukowska Bożena
Department of Biophysics of Environmental Pollution, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska Str. 141/143, 90-236 Lodz, Poland.
Department of Translational Research, Nofer Institute of Occupational Medicine, Teresy Str. 8, 91-348 Lodz, Poland.
Int J Mol Sci. 2024 Nov 28;25(23):12786. doi: 10.3390/ijms252312786.
The aim of the present study was to investigate the concentration- and size-dependent effects of non-functionalized polystyrene nanoparticles (PS-NPs) of varying diameters (29 nm, 44 nm, and 72 nm) on specific epigenetic modifications and gene expression profiles related to carcinogenesis in human peripheral blood mononuclear cells (PBMCs) in vitro. This in vitro human-cell-based model is used to investigate the epigenetic effect of various environmental xenobiotics. PBMCs were exposed to PS-NPs at concentrations ranging from 0.001 to 100 µg/mL for 24 h period. The analysis encompassed epigenetic DNA modifications, including levels of 5-methyl-2'-deoxycytidine (5-mdC) and 5-(hydroxymethyl)-2'-deoxycytidine (5-hmdC), as well as the levels of 2'-deoxyuridine (dU) and 5-(hydroxymethyl)-2'-deoxyuridine (5-hmdU) by mass spectrometry methods, methylation in the promoter regions of selected tumor suppressor genes (P53), (P16), and (P21) and proto-oncogenes (, , ), along with the expression profile of the indicated genes by real-time PCR assays. The results obtained revealed no significant changes in global DNA methylation/demethylation levels in PBMCs after short-term exposure to non-functionalized PS-NPs. Furthermore, there were no changes observed in the level of dU, a product of cytosine deamination. However, the level of 5-hmdU, a product of both 5-hmdC deamination and thymine oxidation, was increased at the highest concentrations of larger PS-NPs (72 nm). None of the PS-NPs caused a change in the methylation pattern of the promoter regions of the , , , , and genes. However, gene profiling indicated that PS-NPs with a diameter of 29 nm and 44 nm altered the expression of the gene. The smallest PS-NPs with a diameter of 29 nm increased the expression of the gene at a concentration of 10 µg/mL, while PS-NPs with a diameter of 44 nm did so at a concentration of 100 µg/mL. An increase in the expression of the gene was also observed when PBMCs were exposed to PS-NPs with 29 nm in diameter at the highest concentration. The observed effect depended on both the concentration and the size of the PS-NPs.
本研究的目的是调查不同直径（29纳米、44纳米和72纳米）的非功能化聚苯乙烯纳米颗粒（PS-NPs）在体外对人外周血单核细胞（PBMCs）中与致癌作用相关的特定表观遗传修饰和基因表达谱的浓度和尺寸依赖性影响。这种基于体外人类细胞的模型用于研究各种环境异生物素的表观遗传效应。将PBMCs暴露于浓度范围为0.001至100微克/毫升的PS-NPs中24小时。分析包括表观遗传DNA修饰，包括5-甲基-2'-脱氧胞苷（5-mdC）和5-（羟甲基）-2'-脱氧胞苷（5-hmdC）的水平，以及通过质谱法检测的2'-脱氧尿苷（dU）和5-（羟甲基）-2'-脱氧尿苷（5-hmdU）的水平，选定的肿瘤抑制基因（P53）、（P16）和（P21）以及原癌基因（、、）启动子区域的甲基化情况，以及通过实时PCR测定法检测上述基因的表达谱。所获得的结果显示，短期暴露于非功能化PS-NPs后，PBMCs中的整体DNA甲基化/去甲基化水平没有显著变化。此外，未观察到胞嘧啶脱氨产物dU水平的变化。然而，在较大尺寸PS-NPs（72纳米）的最高浓度下，5-羟甲基胞嘧啶脱氨和胸腺嘧啶氧化的产物5-hmdU水平升高。没有一种PS-NPs导致、、、、和基因启动子区域的甲基化模式发生变化。然而，基因分析表明，直径为29纳米和44纳米的PS-NPs改变了基因的表达。直径为29纳米的最小PS-NPs在浓度为10微克/毫升时增加了基因的表达，而直径为44纳米的PS-NPs在浓度为100微克/毫升时增加了该基因的表达。当PBMCs暴露于最高浓度的直径为29纳米的PS-NPs时，也观察到基因表达增加。观察到的效应取决于PS-NPs的浓度和尺寸。
