Laser-capture microdissection in prostate cancer research: establishment and validation of a powerful tool for the assessment of tumour-stroma interactions

BJU International
Chitranjan J ShuklaDylan R W Edwards

Abstract

To describe our experience with the optimization and validation of laser-capture microdissection (LCM) for biomarker analysis in prostate tissues. As LCM allows the separation of benign and malignant epithelial structures and stromal elements, it not only allows identification of the source of the biomarker, but might also accentuate gene or protein expression changes by reducing contamination by other cellular elements. In all, 19 fresh-frozen prostate tissue samples were subjected to LCM, with the cDNA being analysed using quantitative polymerase chain reaction for several genes, to identify the optimum number of cells for capture, as well as gene markers assessing for the purity of the captured cells. The localization was further confirmed by in situ hybridization. Prostate-specific antigen (PSA) and cytokeratin 8, were expressed solely by epithelial cells, whereas hepatocyte growth factor (HGF) and tissue inhibitor of metalloproteinases-3 (TIMP3) were expressed only by stromal cells, and the levels of transcripts of these genes were unaltered between benign and malignant tissues. These data suggest that PSA, cytokeratin 8, HGF and TIMP3 are reliable gene markers of purity of epithelial and stromal compartments for LCM of pr...Continue Reading

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Citations

Jul 24, 2013·Briefings in Functional Genomics·Bryan D HudsonGabriela G Loots
Mar 25, 2010·World Journal of Gastroenterology : WJG·Clara Luz SampieriKenneth León-Córdoba
May 23, 2009·The Prostate·Larisa NonnPeter H Gann
Jun 19, 2013·International Journal of Molecular Sciences·Marina RigauAndreas Doll

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