Italian Workers' Compensation Authority (INAIL), Department of Production Plants and Anthropic Settlements (DIPIA) Via Alessandria, 220/E, 00198 Rome, Italy.
University of Rome "La Sapienza"-BCE, Viale del Policlinico 155, 00161, Rome, Italy.
Sci Total Environ. 2014 Feb 15;472:672-80. doi: 10.1016/j.scitotenv.2013.11.092. Epub 2013 Dec 7.
Exposure to inorganic Arsenic (iAs) through drinking water is a major public health problem affecting most countries. iAs has been classified by the International Agency for Research on Cancer as Group 1: "Carcinogenic to humans". Although numerous studies have shown the related adverse effects of iAs, sensitive appropriate biomarkers for studies of environmental epidemiology are still required. The present work aims at investigate the role of microRNAs (miRNAs), powerful negative regulators of gene expression, playing a key role in many physiological and pathological cellular processes, in iAs exposure. To this end, we analyzed miRNA changes in expression profile triggered by iAs exposure in Jurkat cell line. We used microarray technology to profile the expression of miRNAs following 2 μmol/L sodium arsenite treatment at different time points. Moreover, we performed phenotypic analysis of iAs treated cells. Real Time Polymerase Chain Reaction (RT-PCR) was used to validate miRNA microarray data and to assay expression modulation of selected relevant mRNAs. Finally, bioinformatics techniques were applied to reconstruct iAs-relevant molecular pathways and miRNA regulatory networks from the expression data. We report miRNAs modulated after iAs treatment in Jurkat cells. In particular, we highlight 36 miRNAs exhibiting consistent dysregulation and particularly a panel of 8 miRNAs which we also validated by RT-PCR analysis. Computational analysis of lists of putative target genes for these 8 miRNAs points to an involvement in arsenic-response pathways, for a subset of them, that were analyzed by RT-PCR. Furthermore, iAs exposure reveals induction of cell cycle progression and the failure of apoptosis, supporting the idea of iAs carcinogenic activity. Our study provides a list of miRNAs whose expression levels are affected by iAs treatment, corroborating the importance of proceeding with the hunt for specific subset of miRNAs, which can serve as potential biomarkers of iAs effects with useful diagnostic value.
通过饮用水接触无机砷（iAs）是影响大多数国家的主要公共卫生问题。国际癌症研究机构已将 iAs 归类为第 1 组：“对人类致癌”。尽管许多研究表明 iAs 具有相关的不良影响，但仍需要敏感且适当的生物标志物来进行环境流行病学研究。本工作旨在研究微小 RNA（miRNAs）在 iAs 暴露中的作用，miRNAs 是基因表达的强大负调控因子，在许多生理和病理细胞过程中发挥关键作用。为此，我们分析了 Jurkat 细胞系中 iAs 暴露引发的 miRNA 表达谱变化。我们使用微阵列技术在不同时间点分析了 2 μmol/L 亚砷酸钠处理后 miRNA 的表达谱。此外，我们对 iAs 处理的细胞进行了表型分析。实时聚合酶链反应（RT-PCR）用于验证 miRNA 微阵列数据，并测定选定相关 mRNA 的表达调节。最后，应用生物信息学技术从表达数据中重建与 iAs 相关的分子途径和 miRNA 调控网络。我们报告了 Jurkat 细胞中 iAs 处理后调节的 miRNA。特别是，我们强调了 36 个 miRNA 表现出一致的失调，特别是我们也通过 RT-PCR 分析验证的 8 个 miRNA 面板。这些 8 个 miRNA 的假定靶基因列表的计算分析表明，它们参与了砷反应途径，其中一部分通过 RT-PCR 进行了分析。此外，iAs 暴露揭示了细胞周期进程的诱导和细胞凋亡的失败，支持了 iAs 致癌活性的观点。我们的研究提供了一组 miRNA 的表达水平受 iAs 处理影响的列表，证实了继续寻找特定 miRNA 子集的重要性，这些子集可以作为 iAs 效应的潜在生物标志物，具有有用的诊断价值。
