Exploring gene expression signatures of itraconazole for infantile hemangioma endothelial cell in vitro. Exploring gene expression signatures of itraconazole for infantile hemangioma endothelial cell in vitro

In order to understand the mechanisms underlying the effects of itraconazole in infantile hemangioma endothelial cells, we further explored the potential targeting genes of itraconazole in infantile hemangioma endothelial cells in vitro by using a genome-wide gene expression array. In total 172 mRNAs were up-regulated (fold change > 2) and 819 mRNAs were down-regulated (fold change > 2) upon itraconazole treatment in infantile hemangioma endothelial cells. The top signaling pathway closely related to differentially expressed genes upon itraconazole treatment is the cytokine and cytokine receptor pathway. Within this pathway, the mRNA expression of PDGF-D, an important growth factor belonging to the PDGF family, was the most significantly reduced (30 folds) among 991 differentially expressed genes after itraconazole treatment. qRT-PCR and western blotting confirmed the inhibitory effects on PDGF-D levels. Overall design: Infantile hemangioma endothelial cells from 2 infants were treated with 10μM itraconazole or 0.1% DMSO for 48 hours. Total mRNAs were extracted used for mRNA array analysis with Agilent's Human (V2) Gene Expression Microarray (8×60K chip), according to the manufacturer’s instructions.

为阐明伊曲康唑作用于婴幼儿血管瘤内皮细胞的分子机制，本研究通过全基因组基因表达阵列（genome-wide gene expression array），体外探究了伊曲康唑在婴幼儿血管瘤内皮细胞中的潜在靶向基因。经伊曲康唑处理后，婴幼儿血管瘤内皮细胞中共筛选得到172个上调mRNA（倍数变化>2）、819个下调mRNA（倍数变化>2）。与伊曲康唑处理后差异表达基因密切相关的核心信号通路为细胞因子与细胞因子受体通路。在该通路中，属于血小板衍生生长因子家族的重要生长因子PDGF-D的mRNA表达水平，在伊曲康唑处理后的991个差异表达基因中下调幅度最为显著（达30倍）。实时定量聚合酶链式反应（qRT-PCR）与蛋白质印迹（western blotting）实验验证了伊曲康唑对PDGF-D表达的抑制作用。实验整体设计：从2名婴幼儿体内分离得到的血管瘤内皮细胞，分别用10μM伊曲康唑或0.1%二甲基亚砜（DMSO）处理48小时。提取总mRNA后，按照制造商说明书，使用安捷伦（Agilent）人类（V2）基因表达微阵列芯片（8×60K芯片）开展mRNA芯片分析。