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An in vitro method to survey DNA methylation in peripheral blood mononuclear cells (PBMCs) treated by airborne particulate matter (PM 10)

机译:一种体外方法研究经空气传播颗粒物质处理的外周血单核细胞(PBMC)中的DNA甲基化(PM 10

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The International Agency for Research on Cancer (IARC) has defined outdoor air pollution and PM as the human carcinogen (Group 1), which mainly cause an increased risk of lung cancer. Scientists have considered epigenetic modifications as a possible mechanism to deal with adverse effects of air pollution. This study aimed to compare the effect of airborne PM10(PM with aerodynamic diameter ≤10?μm) on in vitro global methylation in human peripheral blood mononuclear cells (PBMCs). PM10was sampled in metropolitan Tehran, the capital of Iran. PBMCs were extracted from whole blood of healthy males and treated with PM10suspension at concentrations of 50–300?μg/mL for 4?h. Untreated cells were used as the negative control. Genomic DNA was extracted from each sample using the DNA blood mini kit according to the manufacturer’s instruction. Moreover, 5-methylsytosine (%5-mC) and 5-hydroxymethylcytosine (%5-hmC) were measured by the enzyme-linked immunosorbent assay (ELISA) method. %5-mC and %5-hmC in each sample was compared with negative control and reported as difference %5-mC and %5-hmC.
机译:国际癌症研究机构(IARC)将室外空气污染和PM定义为人类致癌物(第1组),这主要导致肺癌风险增加。科学家已经将表观遗传修饰视为应对空气污染不利影响的一种可能机制。本研究旨在比较机载PM10(空气动力学直径≤10μm的PM)对人外周血单个核细胞(PBMC)体外总体甲基化的影响。 PM10是在伊朗首都德黑兰进行取样的。从健康男性全血中提取PBMC,并以50–300?μg/ mL的PM10悬浮液处理4?h。未处理的细胞用作阴性对照。根据制造商的说明,使用DNA血液微型试剂盒从每个样品中提取基因组DNA。另外,通过酶联免疫吸附测定(ELISA)法测定了5-甲基肌嘧啶(%5-mC)和5-羟甲基胞嘧啶(%5-hmC)。将每个样品中的%5-mC和%5-hmC与阴性对照进行比较,并报告为差异%5-mC和%5-hmC。

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