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首页> 外文期刊>Endothelium: Journal of endothelial cell research >A novel methodology to probe endothelial differential gene expression profile reveals novel genes.
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A novel methodology to probe endothelial differential gene expression profile reveals novel genes.

机译：一种探测内皮细胞差异基因表达谱的新方法揭示了新基因。

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摘要

Endothelial dysfunction is a major feature of vascular diseases. A practical, minimally invasive method to effectively "probe" gene transcription for an individual patient's endothelium has potential to "customize" assessment for an individual at risk of vascular disease as well as pathophysiologic insight in an in vivo human, clinical context. Published literature lacks a methodology to identify endothelial differential gene expression in individuals with vascular disease. We describe a methodology to do so. The aim of this study was to specifically utilize (a) cutaneous microvascular biopsy, (b) laser capture microdissection, (c) cDNA amplification, (d) suppression subtractive hybridization, (e) high-throughput sequencing techniques, (f) real-time polymerase chain reaction (PCR), and (g) in combination of these methods, to profile differential gene expression in the context of cardiovascular and cerebrovascular disease. Endothelial cells were obtained by laser capture microdissection from a patient and a healthy sibling's microvascular biopsy tissues. Endothelial RNA was extracted, reverse transcribed, and amplified to ds cDNA. Suppression subtractive hybridization was used to establish an endothelial differential gene expression library. Real-time PCR confirmed SERP1, caspase 8, IGFBP7, S100A4, F85, and F147 up-regulation between 1.4- and 3.47-fold. The authors have successfully established a methodology to profile endothelial differential gene expression and identified six differentially expressed genes. This minimally invasive novel method has potential wide application in the customized assessment of many patients suffering vascular diseases.
机译：内皮功能障碍是血管疾病的主要特征。一种有效地针对个体患者的内皮有效“探查”基因转录的实用，微创方法具有潜力，可以“定制”对有血管疾病风险的个体的评估，以及在体内人类临床环境中的病理生理学见解。公开的文献缺乏鉴定具有血管疾病的个体中的内皮细胞差异基因表达的方法。我们描述了这样做的方法。这项研究的目的是专门利用（a）皮肤微血管活检，（b）激光捕获显微切割，（c）cDNA扩增，（d）抑制消减杂交，（e）高通量测序技术，（f）真正的时聚合酶链反应（PCR）和（g）这些方法的组合，以在心血管和脑血管疾病的背景下分析差异基因的表达。通过激光捕获显微切割从患者和健康兄弟姐妹的微血管活检组织中获得内皮细胞。提取内皮RNA，逆转录，并扩增为ds cDNA。使用抑制性消减杂交建立内皮差异基因表达文库。实时PCR证实SERP1，胱天蛋白酶8，IGFBP7，S100A4，F85和F147上调在1.4到3.47倍之间。作者已经成功地建立了描述内皮细胞差异基因表达的方法，并鉴定了六个差异表达基因。这种微创的新方法在许多血管疾病患者的定制评估中具有潜在的广泛应用。

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