Kim Hwan-Young, Kim Hye-Ran, Kang Min-Gu, Trang Nguyen Thi Dai, Baek Hee-Jo, Moon Jae-Dong, Shin Jong-Hee, Suh Soon-Pal, Ryang Dong-Wook, Kook Hoon, Shin Myung-Geun
Department of Laboratory Medicine, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, 160 Ilsimri, Hwasun-eup, Hwasun-gun, Jeollanam-do 519-809, Republic of Korea ; Brain Korea 21 Plus Project, Chonnam National University Medical School, Gwangju, Republic of Korea.
Brain Korea 21 Plus Project, Chonnam National University Medical School, Gwangju, Republic of Korea ; Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Biomed Res Int. 2014;2014:605135. doi: 10.1155/2014/605135. Epub 2014 Jul 10.
This study investigated the profiling of polycyclic aromatic hydrocarbon- (PAH-) induced genotoxicity in cell lines and zebrafish. Each type of cells displayed different proportionality of apoptosis. Mitochondrial DNA (mtDNA) copy number was dramatically elevated after 5-day treatment of fluoranthene and pyrene. The notable deregulated proteins for PAHs exposure were displayed as follows: lamin-A/C isoform 3 and annexin A1 for benzopyrene; lamin-A/C isoform 3 and DNA topoisomerase 2-alpha for pentacene; poly[ADP-ribose] polymerase 1 (PARP-1) for fluoranthene; and talin-1 and DNA topoisomerase 2-alpha for pyrene. Among them, lamin-A/C isoform 3 and PARP-1 were further confirmed using mRNA and protein expression study. Obvious morphological abnormalities including curved backbone and cardiomegaly in zebrafish were observed in the 54 hpf with more than 400 nM of benzopyrene. In conclusion, the change of mitochondrial genome (increased mtDNA copy number) was closely associated with PAH exposure in cell lines and mesenchymal stem cells. Lamin-A/C isoform 3, talin-1, and annexin A1 were identified as universal biomarkers for PAHs exposure. Zebrafish, specifically at embryo stage, showed suitable in vivo model for monitoring PAHs exposure to hematopoietic tissue and other organs.
本研究调查了多环芳烃(PAH)在细胞系和斑马鱼中诱导的基因毒性特征。每种类型的细胞显示出不同比例的凋亡。荧蒽和芘处理5天后,线粒体DNA(mtDNA)拷贝数显著升高。PAHs暴露后显著失调的蛋白质如下:苯并[a]芘的核纤层蛋白A/C异构体3和膜联蛋白A1;并五苯的核纤层蛋白A/C异构体3和DNA拓扑异构酶2-α;荧蒽的聚[ADP-核糖]聚合酶1(PARP-1);以及芘的踝蛋白-1和DNA拓扑异构酶2-α。其中,通过mRNA和蛋白质表达研究进一步证实了核纤层蛋白A/C异构体3和PARP-1。在54小时胚胎期,用超过400 nM的苯并[a]芘处理斑马鱼,观察到明显的形态异常,包括脊柱弯曲和心脏肥大。总之,线粒体基因组的变化(mtDNA拷贝数增加)与细胞系和间充质干细胞中的PAH暴露密切相关。核纤层蛋白A/C异构体3、踝蛋白-1和膜联蛋白A1被确定为PAHs暴露的通用生物标志物。斑马鱼,特别是在胚胎阶段,显示出适用于监测PAHs对造血组织和其他器官暴露的体内模型。