Joseph Ajay, Gnanapragasam Vincent J
Translational Prostate Cancer Group, Hutchison MRC Research Centre, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2011;755:291-300. doi: 10.1007/978-1-61779-163-5_24.
Prognostic markers can improve prediction of the behaviour of a cancer at the point of diagnosis. A key value of any prognostic marker is at the point of tumour diagnosis. In the context of prostate cancer, this implies profiling in the diagnostic formalin-fixed, paraffin-embedded (FFPE) transrectal ultrasound-guided (TRUS) needle biopsy. TRUS needle biopsies commonly contain both stromal and epithelial cells, and malignant glands are found as isolated foci within this tissue. Using the entire biopsy for genetic analysis inevitably results in a significant contamination of malignant cells with benign tissue. This combination of minimal tumour yields and tissue heterogeneity have so far prohibited prognostic transcript and microarray molecular studies in needle biopsies. Laser-capture microdissection (LCM) allows enriched cell populations to be accurately isolated from heterogeneous tissue, hence facilitating analysis of different components from a single tissue sample. Here, we describe its use in isolating tumour cells in archival FFPE prostate needle biopsies and subsequent application for RNA extraction and quantitative real-time PCR (QPCR).
预后标志物可以在癌症诊断时改善对其行为的预测。任何预后标志物的关键价值都体现在肿瘤诊断之时。就前列腺癌而言，这意味着要对诊断性福尔马林固定、石蜡包埋（FFPE）经直肠超声引导（TRUS）穿刺活检组织进行分析。TRUS穿刺活检组织通常同时包含基质细胞和上皮细胞，恶性腺体在该组织中呈孤立灶状分布。使用整个活检组织进行基因分析不可避免地会导致恶性细胞被良性组织大量污染。肿瘤样本量少和组织异质性这两个因素，迄今为止阻碍了对穿刺活检组织进行预后转录组和微阵列分子研究。激光捕获显微切割（LCM）能够从异质性组织中准确分离出富集的细胞群体，从而便于对单个组织样本的不同成分进行分析。在此，我们描述了其在从存档的FFPE前列腺穿刺活检组织中分离肿瘤细胞以及随后用于RNA提取和定量实时PCR（QPCR）方面的应用。
