Transcriptomic Analysis of Human Primary Bronchial Epithelial Cells after Chloropicrin Treatment

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• 作者：Maija Pesonen ; Markus Storvik ; Tarja Kokkola ; Jaana Rys盲 ; Kirsi V盲h盲kangas ; Markku Pasanen
• 刊名：Chemical Research in Toxicology
• 出版年：2015
• 出版时间：October 19, 2015
• 年：2015
• 卷：28
• 期：10
• 页码：1926-1935
• 全文大小：402K
• ISSN：1520-5010

文摘

Chloropicrin is a vaporizing toxic irritant that poses a risk to human health if inhaled, but the mechanism of its toxicity in the respiratory tract is poorly understood. Here, we exposed human primary bronchial epithelial cells (HBEpC) to two concentrations of chloropicrin (10鈥?0 渭M) for 6 or 48 h and used genomic microarray, flow cytometry, and TEM-analysis to monitor cellular responses to the exposures. The overall number of differentially expressed transcripts with a fold-change > 卤 2 compared to controls increased with longer exposure times. The initial response was activation of genes with a higher number of up- (512 by 10 渭M and 408 by 40 渭M chloropicrin) rather than down-regulated transcripts (40 by 10 渭M and 215 by 40 渭M chloropicrin) at 6 h seen with both exposure concentrations. The number of down-regulated transcripts, however, increased with the exposure time. The differentially regulated transcripts were further examined for enriched Gene Ontology Terms (GO) and KEGG-pathways. According to this analysis, the 鈥渞ibosome鈥?and 鈥渙xidative phosphorylation鈥?were the KEGG-pathways predominantly affected by the exposure. The predominantly affected (GO) biological processes were 鈥減rotein metabolic process鈥?including 鈥渢ranslation,鈥?鈥渃ellular protein complex assembly,鈥?and 鈥渞esponse to unfolded protein.鈥?Furthermore, the top pathways, 鈥淣RF2-activated oxidative stress鈥?and 鈥淎h-receptor signaling,鈥?were enriched in our data sets by IPA-analysis. Real time qPCR assay of six selected genes agreed with the microarray analysis. In addition, chloropicrin exposure increased the numbers of late S and/or G2/M-phase cells as analyzed by flow cytometry and induced autophagy as revealed by electron microscopy. The targets identified are critical for vital cellular functions reflecting acute toxic responses and are potential causes for the reduced viability of epithelial cells after chloropicrin exposure.