The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types. Healthy prostate is comprised of fibromuscular stroma surrounding discrete epithelial-lined secretory lumens and a very small population of immune and neuroendocrine cells.
In contrast, areas of prostate cancer have increased dysplastic luminal epithelium with greatly reduced or absent stromal population. Given the profound difference between stromal and epithelial cell types, it is imperative to separate the cell types for any type of downstream molecular analysis. Despite this knowledge, the bulk of gene expression studies compare benign prostate to cancer without micro-dissection, leading to stromal bias in the benign samples. Laser-capture micro-dissection (LCM) is an effective method to physically separate different cell types from a specimen section. The goal of this protocol is to show that RNA can be successfully isolated from LCM-collected human prostatic epithelium and used for downstream gene expression studies such as RT-qPCR and RNAseq.
Journal of visualized experiments : JoVE. 2015 Nov 26*** epublish ***
Giovanni Lugli, Yachana Kataria, Zachary Richards, Peter Gann, Xiaofeng Zhou, Larisa Nonn
Department of Pathology, University of Illinois at Chicago. , Department of Pathology, University of Illinois at Chicago. , Department of Pathology, University of Illinois at Chicago. , Department of Pathology, University of Illinois at Chicago. , Department of Periodontics, University of Illinois at Chicago. , Department of Pathology, University of Illinois at Chicago;