Tumor-associated endothelial cells display GSTP1 and RARbeta2 promoter methylation in human prostate cancer

Journal of Translational Medicine
Amelia C GroverSteven K Libutti

Abstract

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...Continue Reading

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Citations

Mar 18, 2011·Nature Protocols·Jeffrey C HansonMichael R Emmert-Buck
Jan 1, 2016·Clinical & Translational Immunology·Fiorella M SpinelliLaura Alaniz
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Methods Mentioned

BETA
fluorescence activated cell sorting
dissections
PCR
dissection

Software Mentioned

xMD

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