Grover Amelia C, Tangrea Michael A, Woodson Karen G, Wallis Benjamin S, Hanson Jeffrey C, Chuaqui Rodrigo F, Gillespie John W, Erickson Heidi S, Bonner Robert F, Pohida Thomas J, Emmert-Buck Michael R, Libutti Steven K
Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
J Transl Med. 2006 Mar 2;4:13. doi: 10.1186/1479-5876-4-13.
A functional blood supply is essential for tumor growth and proliferation. However, the mechanism of blood vessel recruitment to the tumor is still poorly understood. Ideally, a thorough molecular assessment of blood vessel cells would be critical in our comprehension of this process. Yet, to date, there is little known about the molecular makeup of the endothelial cells of tumor-associated blood vessels, due in part to the difficulty of isolating a pure population of endothelial cells from the heterogeneous tissue environment.
Here we describe the use of a recently developed technique, Expression Microdissection, to isolate endothelial cells from the tumor microenvironment. The methylation status of the dissected samples was evaluated for GSTP1 and RARbeta2 promoters via the QMS-PCR method.
Comparing GSTP1 and RARbeta2 promoter methylation data, we show that 100% and 88% methylation is detected, respectively, in the tumor areas, both in epithelium and endothelium. Little to no methylation is observed in non-tumor tissue areas.
We applied an accurate microdissection technique to isolate endothelial cells from tissues, enabling DNA analysis such as promoter methylation status. The observations suggest that epigenetic alterations may play a role in determining the phenotype of tumor-associated vasculature.
功能性血液供应对肿瘤生长和增殖至关重要。然而，血管向肿瘤募集的机制仍知之甚少。理想情况下，对血管细胞进行全面的分子评估对于我们理解这一过程至关重要。然而，迄今为止，关于肿瘤相关血管内皮细胞的分子组成知之甚少，部分原因是难以从异质组织环境中分离出纯的内皮细胞群体。
在此我们描述了使用一种最近开发的技术——表达显微切割，从肿瘤微环境中分离内皮细胞。通过QMS-PCR方法评估切割样本中GSTP1和RARbeta2启动子的甲基化状态。
比较GSTP1和RARbeta2启动子甲基化数据，我们发现肿瘤区域的上皮和内皮中分别检测到100%和88%的甲基化。在非肿瘤组织区域几乎未观察到甲基化。
我们应用精确的显微切割技术从组织中分离内皮细胞，从而能够进行DNA分析，如启动子甲基化状态分析。这些观察结果表明，表观遗传改变可能在决定肿瘤相关脉管系统的表型中起作用。
