Molecular profiling of angiogenesis markers
Molecular profiling of angiogenesis markers
复制标题
DOI：
10.1016/s0002-9440(10)64154-5
复制
发表时间：
2002-07-01
影响因子：
6
通讯作者：
Senger, DR
中科院分区：
文献类型：
--
作者：
Shih, SC;Robinson, GS;Senger, DR
本研究的目的是开发一种灵敏、简单、广泛适用的检测方法，利用实时定量逆转录-聚合酶链反应（RT-PCR）测量特定mRNA的拷贝数，并确定与血管生成密切相关的基因表达谱。我们测量了一组9个潜在的血管生成标记物从小鼠转基因模型的前列腺癌（TRAMP）和小鼠皮肤模型的血管内皮生长因子（VEGF）驱动的血管生成。在两种模型中，VEGF的表达与编码其他血管生成细胞因子（血管生成素-1和血管生成素-2）、内皮细胞受体酪氨酸激酶（Flt-1、KDR、Tie-1）和内皮细胞粘附分子（VE-钙粘蛋白、PECAM-1）的mRNA的表达相关。与对照组相比，在VEGF高度刺激的真皮中，Ang-2 mRNA转录数增加35倍，PECAM-1和VE-钙粘蛋白增加10倍，Tie-1增加8倍，KDR和Flt-1各增加4倍，Ang-1增加2倍。在VEGF表达较少的皮肤中，所有转录物的数量都相应地减少，表明这七种标志物中的每一种与VEGF的关系。因此，本研究确定了一种高效的方法，用于精确定量与VEGF表达和VEGF诱导的新血管形成相关的一组7种特异性mRNA，并提供了证据表明实时定量RT-PCR提供了一种高度敏感的监测血管生成的策略。
The goal of this study was to develop a sensitive, simple, and widely applicable assay to measure copy numbers of specific mRNAs using real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), and identify a profile of gene expression closely associated with angiogenesis. We measured a panel of nine potential angiogenesis markers from a mouse transgenic model of prostate adenocarcinoma (TRAMP) and a mouse skin model of vascular endothelial growth factor (VEGF)-driven angiogenesis. In both models, expression of VEGF correlated with expression of mRNAs encoding other angiogenic cytokines (angiopoietin-1 and angiopoietin-2), endothelial cell receptor tyrosine kinases (Flt-1, KDR, Tie-1), and endothelial cell adhesion molecules (VE-cadherin, PECAM-1). Relative to control, in dermis highly stimulated by VEGF, the Ang-2 mRNA transcript numbers increased 35-fold, PECAM-1 and VE-cadherin increased 10-fold, Tie-1 increased 8-fold, KDR and Flt-1 each increased 4-fold, and Ang-1 increased 2-fold. All transcript numbers were correspondingly reduced in skin with less VEGF expression, indicating a relationship of each of these seven markers with VEGF. Thus, this study identifies a highly efficient method for precise quantification of a panel of seven specific mRNAs that correlate with VEGF expression and VEGF-induced neovascularization, and it provides evidence that real-time quantitative RT-PCR offers a highly sensitive strategy for monitoring angiogenesis.