Han Tae-Su, Oshima Masanobu
Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan.
Methods Mol Biol. 2018;1725:143-153. doi: 10.1007/978-1-4939-7568-6_12.
Cancer tissues are composed of various cell types including cancer cells, cancer-associated fibroblasts (CAFs), tumor-associated macrophages (TAMs) and endothelial cells. These surrounding stromal cells form the tumor microenvironment, partly through the inflammatory response, which plays an important role in the development and malignant progression of cancer. It is therefore important to examine the expression profiles and protein modifications of each cellular component independently to decipher the interaction between the tumor cells and the microenvironment. We herein describe a protocol for laser microdissection, which allows for the individual cellular compartments to be collected separately. This will allow us to perform real-time RT-PCRs and microarray analyses of specific cell types in tumor tissues.
癌组织由多种细胞类型组成,包括癌细胞、癌症相关成纤维细胞(CAFs)、肿瘤相关巨噬细胞(TAMs)和内皮细胞。这些周围的基质细胞部分通过炎症反应形成肿瘤微环境,炎症反应在癌症的发生和恶性进展中起重要作用。因此,独立检测每个细胞成分的表达谱和蛋白质修饰,以解读肿瘤细胞与微环境之间的相互作用非常重要。我们在此描述一种激光显微切割方案,该方案可分别收集各个细胞区室。这将使我们能够对肿瘤组织中的特定细胞类型进行实时逆转录聚合酶链反应(RT-PCR)和微阵列分析。