Valproic Acid-Induced Deregulation In Vitro of Genes Associated In Vivo with Neural Tube Defects
Valproic Acid-Induced Deregulation In Vitro of Genes Associated In Vivo with Neural Tube Defects
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DOI：
10.1093/toxsci/kfp002
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发表时间：
2009-03-01
影响因子：
3.8
通讯作者：
Stigson, Michael
中科院分区：
文献类型：
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作者：
Jergil, Mans;Kultima, Kim;Stigson, Michael
探索了体外系统寻找发育毒性的分子靶点和标记物的效用，使用微阵列检测多能小鼠胚胎癌细胞系P19中易受致畸剂丙戊酸（VPA）失调影响的基因。使用CodeLink UniSet I Mouse 20 K Expression Bioarrays对在不存在或存在1、2.5或10 mM VPA的情况下培养1.5、6或24 h的P19细胞中提取的总RNA进行重复的微阵列分析。中度F检验显示2972个（p < 10（-3））阵列探针（过滤基因列表的19.4%）的VPA反应显著，其中421个在所有时间点均显著。在表达下调的VPA靶基因的核心子集中，（68个基因）或上调（125个基因）（p < 10（-7））VPA暴露1.5和6小时后，下调基因中的生物学过程基因本体论术语转录调控显著富集，上调基因中的凋亡显著富集，两个下调的基因（Folr 1和Gtf 2 i）具有包括露脑畸形的敲除表型，露脑畸形是VPA在小鼠中诱导的主要畸形。P19细胞中VPA诱导的基因表达反应表明，从遗传小鼠模型中已知的与神经管缺陷相关的200个基因中，约有30%可能是潜在的VPA靶基因，这表明多个基因的联合失调可能是VPA致畸性的机制。还鉴定了与VPA的其他已知效应（组蛋白脱乙酰酶抑制、G（1）期细胞周期阻滞、诱导细胞凋亡）相关的基因表达反应。这项研究表明，体外致畸化合物的毒性基因组学反应可能与已知的体外和体内效应相关，短期（
The utility of an in vitro system to search for molecular targets and markers of developmental toxicity was explored, using microarrays to detect genes susceptible to deregulation by the teratogen valproic acid (VPA) in the pluripotent mouse embryonal carcinoma cell line P19. Total RNA extracted from P19 cells cultured in the absence or presence of 1, 2.5, or 10mM VPA for 1.5, 6, or 24 h was subjected to replicated microarray analysis, using CodeLink UniSet I Mouse 20K Expression Bioarrays. A moderated F-test revealed a significant VPA response for 2972 (p < 10(-3)) array probes (19.4% of the filtered gene list), 421 of which were significant across all time points. In a core subset of VPA target genes whose expression was downregulated (68 genes) or upregulated (125 genes) with high probability (p < 10(-7)) after both 1.5 and 6 h of VPA exposure, there was a significant enrichment of the biological process Gene Ontology term transcriptional regulation among downregulated genes, and apoptosis among upregulated, and two of the downregulated genes (Folr1 and Gtf2i) have a knockout phenotype comprising exencephaly, the major malformation induced by VPA in mice. The VPA-induced gene expression response in P19 cells indicated that similar to 30% of the similar to 200 genes known from genetic mouse models to be associated with neural tube defects may be potential VPA targets, suggestive of a combined deregulation of multiple genes as a possible mechanism of VPA teratogenicity. Gene expression responses related to other known effects of VPA (histone deacetylase inhibition, G(1)-phase cell cycle arrest, induction of apoptosis) were also identified. This study indicates that toxicogenomic responses to a teratogenic compound in vitro may correlate with known in vitro and in vivo effects, and that short-time (