Use of immuno-LCM to identify the in situ expression profile of cellular constituents of the tumor microenvironment.

摘要: Expression profiling using microarrays has become an essential tool for interrogating tumor biology. However, of whole RNA reflects both and host cells, making it difficult to dissect molecular events within specific cellular compartments in the microenvironment. We developed optimized a simple, rapid technique combining immunohistochemistry laser-capture microdissection (immuno-LCM) purify cell populations from microenvironment followed by isolation amplification microarray analysis. Using this methodology, we were able elucidate situ expression profile pure cells endothelial ovarian tumors with brisk immune infiltrates. This not only increased specificity isolated populations, eliminating genes expressed surrounding but also sensitivity analysis, allowing detection low that detected arrays. Pathway analysis identified distinct activation signaling pathways converging on NF-kappaB, as compared cultured lines, which primarily metabolic. Profiling vascular revealed most known panendothelial endothelial-specific markers, unveiled myeloid-monocytic lineage. propose immuno-LCM coupled transcriptional is convenient dissecting complex biological systems such