Kluiver Joost, Slezak-Prochazka Izabella, van den Berg Anke
Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
Methods Mol Biol. 2013;971:265-76. doi: 10.1007/978-1-62703-269-8_15.
MicroRNAs (miRNAs) play important roles in development, differentiation, homeostasis, and also in diseases such as lymphoma. This chapter describes methods to study the role of miRNAs in lymphoma. First, we describe a multiplex RT reaction followed by qPCR that can be used to determine differential expression of candidate miRNAs. Second, we provide a protocol for stable overexpression of miRNAs using lentiviral-based ectopic expression systems. Third, we describe a straightforward assay to determine whether the candidate miRNA may function as an oncogene or a tumor suppressor gene in lymphomagenesis.
微小RNA（miRNA）在发育、分化、体内平衡以及淋巴瘤等疾病中发挥着重要作用。本章介绍了研究miRNA在淋巴瘤中作用的方法。首先，我们描述了一种多重逆转录反应，随后进行定量聚合酶链反应（qPCR），可用于确定候选miRNA的差异表达。其次，我们提供了一种使用基于慢病毒的异位表达系统稳定过表达miRNA的方案。第三，我们描述了一种直接的检测方法，以确定候选miRNA在淋巴瘤发生过程中是否可能作为癌基因或肿瘤抑制基因发挥作用。
