免疫组化检测福尔马林固定石蜡包埋组织中伴侣介导的自噬标志物 LAMP2A 和 HSPA8。

Immunohistochemical Detection of the Chaperone-Mediated Autophagy Markers LAMP2A and HSPA8 in Formalin-Fixed and Paraffin-Embedded Tissues.

机构信息

Institute of Pathology, University of Bern, Bern, Switzerland.

Department of Laboratory Medicine and Pathology, Institute of Pathology, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.

出版信息

Methods Mol Biol. 2023;2566:141-147. doi: 10.1007/978-1-0716-2675-7_11.

DOI:10.1007/978-1-0716-2675-7_11

PMID:36152248

Abstract

Autophagy is crucial for maintaining cellular homeostasis and its deregulation is involved in disease development, including cancer. The key players of chaperone-mediated autophagy (CMA), a particular selective subtype of autophagy, are HSPA8 and LAMP2A. Both proteins can be immunohistochemically detected in formalin-fixed paraffin-embedded (FFPE) tissue. LAMP2A is frequently overexpressed in a variety of cancers where it likely supports cancer cell survival and resistance to anti-cancer therapies in a context-dependent manner. Here we present the immunohistochemical staining protocol of antibodies against LAMP2A and HSPA8, using an automated staining system, suitable for routine diagnostics. Additionally, we also suggest a staining evaluation method.

摘要

自噬对于维持细胞内环境稳定至关重要，其失调与疾病的发展有关，包括癌症。伴侣介导的自噬（CMA）的关键蛋白是 HSPA8 和 LAMP2A，这是自噬的一种特殊选择性亚型。这两种蛋白都可以用福尔马林固定石蜡包埋（FFPE）组织中的免疫组织化学方法检测到。LAMP2A 在多种癌症中经常过表达，它可能以依赖于上下文的方式支持癌细胞的存活和对癌症治疗的耐药性。在这里，我们使用自动化染色系统介绍了针对 LAMP2A 和 HSPA8 的抗体的免疫组织化学染色方案，该方案适用于常规诊断。此外，我们还提出了一种染色评估方法。