Kim Tae-Hoon, Shin Seung-Woo, Park Jong-Sook, Park Choon-Sik
Genome Research Center for Allergy and Respiratory Disease, Soonchunhyang University Bucheon Hospital, 1174, Jung-Dong, Wonmi-Ku, Bucheon, Kyeonggi-Do, 420-020, South Korea.
Environ Toxicol. 2015 Mar;30(3):293-300. doi: 10.1002/tox.21906. Epub 2013 Sep 11.
Environmental particles are believed to provoke airway inflammation in susceptible individuals by stimulating epithelial cells to release mediators that exacerbate lung diseases. Here, we sought to identify genes expressed throughout the genome by epithelial cells stimulated with TiO2 particles. A human bronchial epithelial cell line, BEAS-2B, was stimulated with or without 40 µg TiO2 for 2 h. RNA was purified from cells and subjected to microarray analysis. Genes exhibiting more than a twofold change in RNA expression were selected. Candidate genes were then analyzed using bioinformatics tools, including pathway, ontology, and network analyses. ITGAV mRNA expression levels were measured in BEAS-2B cells using real-time polymerase chain reaction. Among 37,803 genes, 92 genes displayed more than a twofold change in mRNA levels according to the microarray analysis; 87 genes were upregulated while five genes were downregulated. The 92 genes were classified based on functional annotation using a protein information resource database search for biological processes and a pathway search using the KEGG pathway database. These genes are related to macromolecule biosynthesis, metabolic processes and, in particular, RNA metabolism. When genes with more than a threefold change were analyzed, KIF11, ITGAV, SEMA3C, IBTK, and DEK were selected as candidate genes induced by TiO2 -stimulated BEAS-2B cells. To validate these results, BEAS-2B cells stimulated with 40 µg TiO2 expressed threefold higher ITGAV mRNA levels compared to those without TiO2 particle stimulation. We conclude that KIF11, ITGAV, SEMA3C, IBTK, and DEK are candidate genes expressed by epithelial cells when stimulated with TiO2 particles.
环境颗粒物被认为通过刺激上皮细胞释放加剧肺部疾病的介质，从而在易感个体中引发气道炎症。在此，我们试图鉴定经二氧化钛颗粒刺激的上皮细胞在整个基因组中表达的基因。用人支气管上皮细胞系BEAS-2B，在有或无40μg二氧化钛的情况下刺激2小时。从细胞中纯化RNA并进行微阵列分析。选择RNA表达变化超过两倍的基因。然后使用生物信息学工具分析候选基因，包括通路、本体和网络分析。使用实时聚合酶链反应测量BEAS-2B细胞中ITGAV mRNA的表达水平。根据微阵列分析，在37,803个基因中，有92个基因的mRNA水平变化超过两倍；87个基因上调，5个基因下调。使用蛋白质信息资源数据库搜索生物学过程并使用KEGG通路数据库进行通路搜索，基于功能注释对这92个基因进行分类。这些基因与大分子生物合成、代谢过程特别是RNA代谢有关。当分析变化超过三倍的基因时，选择KIF11、ITGAV、SEMA3C、IBTK和DEK作为二氧化钛刺激的BEAS-2B细胞诱导的候选基因。为了验证这些结果，与未用二氧化钛颗粒刺激的细胞相比，用40μg二氧化钛刺激的BEAS-2B细胞表达的ITGAV mRNA水平高两倍。我们得出结论，KIF11、ITGAV、SEMA3C、IBTK和DEK是上皮细胞在受到二氧化钛颗粒刺激时表达的候选基因。
