Grossfeld G D, Shi S R, Ginsberg D A, Rich K A, Skinner D G, Taylor C R, Cote R J
Department of Pathology, University of Southern California School of Medicine, Los Angeles, USA.
J Histochem Cytochem. 1996 Jul;44(7):761-6. doi: 10.1177/44.7.8675997.
Thrombospondin-1 (TSP) is a 450-KD glycoprotein that was initially discovered in the platelet alpha-granule. It now appears that TSP is intimately involved in the regulation of a variety of cellular functions and cell-to-cell interactions. Recently, it has been demonstrated that TSP functions as a p53-dependent inhibitor of angiogenesis in cultured fibroblasts from Li-Fraumeni patients and therefore may be an important factor involved with tumor invasion and metastasis. It has previously been demonstrated that TSP can be detected in frozen tissue sections by immunohistochemical methods. Our objective in this study was to determine the optimal antigen retrieval (AR) protocol for detection of TSP in formalin-fixed, paraffin-embedded tissue by using tissue sections from patients with invasive transitional cell carcinoma of the bladder. The optimal AR protocol was determined utilizing a variety of heating conditions and antigen retrieval buffers. Our results demonstrate that TSP can be reliably detected in paraffin-embedded tissue by immunohistochemical techniques that utilize AR with high-temperature microwave heating and a low-pH Tris-HCI buffer. The importance of this method is that it allows the reliable detection of TSP in archival tissue. This should facilitate further investigation into TSP's role in the regulation of cellular processes, including its influence on tumor angiogenesis and metastasis.
血小板反应蛋白-1（TSP）是一种450千道尔顿的糖蛋白，最初在血小板α颗粒中被发现。现在看来，TSP密切参与多种细胞功能的调节以及细胞间相互作用。最近，已证明TSP在李-弗劳梅尼综合征患者的培养成纤维细胞中作为一种p53依赖性血管生成抑制剂发挥作用，因此可能是参与肿瘤侵袭和转移的一个重要因素。此前已证明，通过免疫组织化学方法可在冷冻组织切片中检测到TSP。本研究的目的是通过使用膀胱浸润性移行细胞癌患者的组织切片，确定在福尔马林固定、石蜡包埋组织中检测TSP的最佳抗原修复（AR）方案。利用多种加热条件和抗原修复缓冲液确定了最佳AR方案。我们的结果表明，通过使用高温微波加热和低pH Tris-HCl缓冲液进行AR的免疫组织化学技术，可在石蜡包埋组织中可靠地检测到TSP。该方法的重要性在于它能在存档组织中可靠地检测到TSP。这应有助于进一步研究TSP在细胞过程调节中的作用，包括其对肿瘤血管生成和转移的影响。
