Immunohistochemical detection of thrombospondin-1 in formalin-fixed, paraffin-embedded tissue
Immunohistochemical detection of thrombospondin-1 in formalin-fixed, paraffin-embedded tissue
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DOI：
10.1177/44.7.8675997
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发表时间：
1996-07-01
影响因子：
3.2
通讯作者：
Cote, RJ
中科院分区：
文献类型：
--
作者：
Grossfeld, GD;Shi, SR;Cote, RJ
血小板反应蛋白-1（TSP）是一种分子量为450-KD的糖蛋白，最初发现于血小板α颗粒。目前看来，TSP密切参与调节多种细胞功能和细胞间相互作用。最近，已经证明TSP在Li-Fraumeni患者的培养成纤维细胞中作为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重要因素。以前已经证明，TSP可以通过免疫组化方法在冷冻组织切片中检测到，我们在这项研究中的目的是确定最佳的抗原修复（AR）协议检测TSP在福尔马林固定，石蜡包埋组织中使用组织切片浸润性膀胱移行细胞癌患者。利用各种加热条件和抗原修复缓冲液确定最佳AR方案。我们的研究结果表明，TSP可以可靠地检测石蜡包埋组织的免疫组织化学技术，利用AR与高温微波加热和低pH值的Tris-HCl缓冲液。该方法的重要性在于其允许可靠地检测存档组织中的TSP。这将有助于进一步研究TSP在调节细胞过程中的作用，包括其对肿瘤血管生成和转移的影响。
Thrombospondin-l (TSP) is a 450-KD glycoprotein that was initially discovered in the platelet a-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-HCl 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.