Radiation Immuno-Oncology Group, Center for Translational Cancer Research Technische Universität München (TranslaTUM), Department of Radiation Oncology, Klinikum rechts der Isar, Technische Universität München (TUM), Munich, Germany.
John van Geest Cancer Research Centre, School of Science and Technology, Nottingham Trent University, Nottingham, UK.
Methods Mol Biol. 2023;2693:307-324. doi: 10.1007/978-1-0716-3342-7_23.
The major stress-inducible 70 kDa heat shock (stress) protein 70 (Hsp70) is frequently overexpressed in highly aggressive tumor cells and thus might serve as a tumor-specific biomarker of aggressive disease and/or therapeutic resistance. We have previously shown that, in contrast to normal cells, tumor cells present Hsp70 on their plasma membrane. In order to elucidate the role of intracellular, membrane-bound and extracellular Hsp70 as a potential tumor biomarker in cancer, herein we describe protocols for the staining of cytosolic Hsp70 in tumor formalin-fixed paraffin-embedded (FFPE) sections from patients with glioblastoma multiforme using immunohistochemistry, for detecting the expression of plasma membrane-bound Hsp70 by a range of cancer-derived cells using multi-parametric flow cytometry using the cmHsp70.1 monoclonal antibody (mAb) and for the measurement of free and vesicular-associated Hsp70 in the circulation of patients with cancer using a unique enzyme-linked immunosorbent assay (ELISA).
主要的应激诱导型 70kDa 热休克（应激）蛋白 70（Hsp70）在高度侵袭性肿瘤细胞中过度表达，因此可能作为侵袭性疾病和/或治疗抵抗的肿瘤特异性生物标志物。我们之前已经表明，与正常细胞相比，肿瘤细胞在其质膜上呈现 Hsp70。为了阐明细胞内、膜结合和细胞外 Hsp70 作为癌症中潜在肿瘤标志物的作用，本文描述了使用免疫组织化学在来自多形性胶质母细胞瘤患者的肿瘤福尔马林固定石蜡包埋（FFPE）切片中染色细胞质 Hsp70 的方案，用于使用 cmHsp70.1 单克隆抗体（mAb）通过多种癌症衍生细胞检测质膜结合 Hsp70 的表达，以及使用独特的酶联免疫吸附试验（ELISA）测量癌症患者循环中的游离和囊泡相关 Hsp70。
