Department of Pathology & Microbiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PE, C1A 4P3, Canada.
Ecotoxicology. 2011 Nov;20(8):1765-72. doi: 10.1007/s10646-011-0709-y. Epub 2011 Jun 18.
The aim of this study is to investigate the effects of the pesticides/polycyclic aromatic hydrocarbon mixture on aryl hydrocarbon receptor (AhR), p53 and ubiquitin mRNA level in haemocytes of Mya arenaria exposed to a mixture of chlorothalonil, mancozeb and benzo[a]pyrene (BaP) for 48 and 72 h. AhR, p53 and ubiquitin gene expression levels were quantified using quantitative Real-time PCR. For robust and accurate quantification of transcripts, suitable housekeeping genes were selected from four sets of ribosomal and elongation factors transcripts previously sequenced from Mya arenaria using geNorm open source software. Quantitative Real-time PCR data exhibited a significantly high expression of AhR after 72 h of exposure (P ≤ 0.05). p53 gene expression seems to be up-regulated by the mixture after 48 h, however not significantly; but the level of p53 mRNA is down-regulated by the xenobiotics between 48 and 72 h after exposure. This study postulates that AhR mRNA levels could be used as an indicator of the exposure of clams' haemocytes to a mixture of xenobiotics such as chlorothalonil, mancozeb and BaP. However, further studies have to be pursued in order to unravel the molecular mechanisms involved in the p53 signaling pathway.
本研究旨在探讨农药/多环芳烃混合物对暴露于百菌清、代森锰锌和苯并[a]芘混合物中的贻贝血红细胞中芳香烃受体 (AhR)、p53 和泛素 mRNA 水平的影响,暴露时间为 48 和 72 小时。采用实时定量 PCR 法对 AhR、p53 和泛素基因表达水平进行定量。为了对转录物进行稳健和准确的定量,使用 geNorm 开源软件从先前从贻贝中测序的四组核糖体和延伸因子转录物中选择了合适的管家基因。实时定量 PCR 数据显示,暴露 72 小时后 AhR 的表达显著升高 (P ≤ 0.05)。混合物在 48 小时后似乎上调了 p53 基因的表达,但不显著;但在暴露后 48 至 72 小时之间,p53 mRNA 的水平被外源化学物质下调。本研究假设 AhR mRNA 水平可作为贻贝血红细胞暴露于百菌清、代森锰锌和苯并[a]芘等外源混合物的指标。然而,为了阐明 p53 信号通路涉及的分子机制,还需要进一步研究。