Added value of stress related gene inductions in HepG2 cells as effect measurement in monitoring of air pollution

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• 作者：Ingrid Nobels^a ; ^{ingrid.nobels@ua.ac.be} ; Caroline Vanparys^a ; Rosette Van den Heuvel^b ; Jordy Vercauteren^c ; Ronny Blust^a
• 关键词：Air pollution ; Reporter assay ; Stress related endpoints
• 刊名：Atmospheric Environment
• 出版年：2012
• 期刊代码：11_13522310
• 类别：env
• 出版时间：August, 2012
• 卷：55
• 期：Complete
• 页码：154-163
• 文件大小：602 K

摘要

In this study we studied the effects of particulate matter samples (PM) through gene expression analysis in a routine air quality monitoring campaign by the Flemish Environment Agency (VMM, Belgium). We selected a human hepatoma (HepG2) multiple endpoint reporter assay for targeted stress related endpoint screening. Organic extracts of air samples (total suspended particles, TSP) were collected during one year in an industrial, urban and background location in Flanders, Belgium. Simultaneously, meteorological conditions (temperature, wind speed and precipitation) and particulate matter size聽鈮ぢ?0聽渭M (PM₁₀), organic (OC), elemental (EC) and total (TC) carbon were monitored and air samples were collected for chemical analysis (11 PAHs). Correlations between the induction of the different stress genes and the chemical pollutants were analysed.

Exposure of HepG2 cells to daily air equivalents (20聽m³) of organic TSP extracts revealed the dominant induction of the xenobiotic response element (Xre) and phase I (Cyp1A1) and phase II (GstYa) biotransformation enzymes. Additional effects were the induction of c-Fos, a proto-oncogen and Gadd45, a marker for cell cycle disturbance and responsive to genotoxic compounds. Inductions of other relevant pathways, such as sequestration of heavy metals, retinoids response, protein misfolding and increased cAMP levels were measured occasionally.

A significant correlation was found between the genes Cyp1A1 (a typical marker for presence of PAHs and dioxin like compounds), c-Fos, Gadd45, (responsive to DNA damaging compounds) and the amount of PM₁₀ and elemental carbon (EC) whereas no correlation was found between these genes and total PAHs content. This may suggest that the observed induction of Cyp1A1 and DNA damage related genes was provoked (partially) by other particle bound compounds (e.g. pesticides, PCBs, brominated flame retardants, dioxins, 鈥?, than PAHs. The contribution of particle bound compounds, other than PAHs might be important to take into account in risk evaluation of air pollution.