Abstract
The association of dioxin exposure with increased morbidity or mortality of chronic cardiovascular diseases (CVDs) has been established by many epidemiological studies. However, the precise global gene expression alterations caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the cardiovascular system need to be further elucidated. In this study, we profiled the gene expression of human umbilical vein endothelial cells (HUVECs) exposed to different concentrations of TCDD by high-throughput sequencing. Expression of 1,838 genes was changed significantly after TCDD stimulation. The FunDO analysis suggested that some CVDs were highly associated with TCDD treatment, including atherosclerosis, thromboangiitis obliterans, pulmonary arterial hypertension (PAH), and hypertension. KEGG pathway analysis showed that many genes in the signaling pathways of vascular smooth muscle contraction and apoptosis were altered distinctly. In addition, we revealed evidence regarding the gene network changes of chronic CVDs including atherosclerosis, thrombosis, myocardial infarction (MI), hypertension, and PAH in TCDD-exposed HUVECs. We found that gene expression of β1-adrenoceptors (ADRB1), β2-adrenoceptors (ADRB2), endothelin-converting enzyme 1 (ECE1), and endothelin-1 gene (EDN1) that are involved in the blood pressure regulation pathway decreased apparently under TCDD treatment. Moreover, the transcripts of interleukin 1 beta (IL-1β) and tumor necrosis factor α (TNFα), which are related to atherosclerosis, were up-regulated by TCDD stimulation. In addition, the transcripts of Homo sapiens collagen, type IV, alpha 1 (COL4A1), and isoforms that trigger the MI pathway were up-regulated after TCDD exposure. Finally, we found enhanced platelet-derived growth factor (PDGF) and signal transducer and activator of transcription 5 (Stat5) expression with TCDD treatment in endothelial cells, which are involved in PAH induced by vascular injury.
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Acknowledgments
This work was supported by grants from the Ministry of Science and Technology of China (2012CB945100, 2012BAK01B00), and the National Natural Science Foundation of China (81030004, 31371154), NSFC-CIHR joint grant (NSFC81161120538 and CIHR-CCI117951), The Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-EW-R-09). Y.Y. was supported by the One Hundred Talents Program of the Chinese Academy of Sciences (2010OHTP10). He is a Fellow at the Jiangsu Collaborative Innovation Center for Cardiovascular Disease Translational Medicine.
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12012_2014_9279_MOESM1_ESM.xls
Supplementary file 1: Table S1. Primers used in qRT-PCR to verify the key genes expression of the AHR pathway. (XLS 17 kb)
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Supplementary file 2: Fig. S1. The cell viabilities of HUVECs treated with different concentrations of TCDD. The HUVECs were grown in medium supplemented with the indicated concentration of TCDD(0, 5, 10, 20, 40, and 80 nM)for 24 h prior to analyzing the cell viability (n = 6). Each point was represented by mean ± SEM. (TIFF 26317 kb)
12012_2014_9279_MOESM3_ESM.xls
Supplementary file 3: Table S2. Genes differentially expressed in TCDD-treated HUVECs. About 23,119 genes with average FPKM ≥ 0.01 in TCDD-exposed HUVECs at least 2 of the 4 treatments were selected to analyze the gene expression profile; 1838 genes with absolute fold change ≥ 1.5 and P < 0.05 were considered as differentially expressed genes in this study. (XLS 666 kb)
12012_2014_9279_MOESM4_ESM.tif
Supplementary file 4: Fig. S2. Heat-map hierarchical clustering of the differentially expressed genes. Heat-map images for the 1,838 differentially expressed genes were shown. Columns and rows in the heat-maps represented samples and genes, respectively. Sample names were displayed below the heat-maps. Color scale indicated gene expression. Red and blue indicated genes with high and low expression, respectively. (TIFF 27645 kb)
12012_2014_9279_MOESM5_ESM.xls
Supplementary file 5: Table S3. TCDD-responsive genes assigned to the principal categories of endothelial function. The genes expression altered by different doses of TCDD was presented as log 2 values. P < 0.05 was considered as statistical significance. (XLS 30 kb)
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Yu, Y., Qin, J., Chen, D. et al. Chronic Cardiovascular Disease-Associated Gene Network Analysis in Human Umbilical Vein Endothelial Cells Exposed to 2,3,7,8-Tetrachlorodibenzo-p-dioxin. Cardiovasc Toxicol 15, 157–171 (2015). https://doi.org/10.1007/s12012-014-9279-6
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DOI: https://doi.org/10.1007/s12012-014-9279-6