Reeves J R, Cooke T G, Fenton-Lee D, McNicol A M, Ozanne B W, Richards R C, Walsh A
Department of Surgery, University of Glasgow, Glasgow Royal Infirmary, United Kingdom.
J Histochem Cytochem. 1994 Mar;42(3):307-14. doi: 10.1177/42.3.8308248.
We developed a sensitive EGF receptor detection method for frozen tissue sections using biotinylated EGF as the primary reagent. The method was directly compared with an immunohistochemical technique based on an anti-EGF receptor monoclonal antibody (MAb EGFR1) in normal human and rat tissues and in human tumors. The method was more sensitive than a previously published biotinylated EGF-based technique. In normal human tissues and in 37 of the 50 tumors, the binding pattern mirrored that of positive staining with EGFR1. Five further tumors showed weak immunoreactivity, but in these no binding of biotinylated EGF was detected. The remaining eight tumors were negative by both techniques. The discordant cases may reflect a lower level of sensitivity of the ligand-binding technique or, alternatively, abnormal receptors may have been expressed in these tissues. EGF receptors could be detected in rat liver with biotinylated EGF but not with the antibody, indicating the usefulness of the ligand-based technique in cross-species studies.
我们开发了一种用于冰冻组织切片的灵敏的表皮生长因子(EGF)受体检测方法,该方法以生物素化的EGF作为主要试剂。在正常人体组织、大鼠组织以及人类肿瘤组织中,将该方法与基于抗EGF受体单克隆抗体(MAb EGFR1)的免疫组织化学技术进行了直接比较。该方法比先前发表的基于生物素化EGF的技术更为灵敏。在正常人体组织以及50个肿瘤中的37个肿瘤中,生物素化EGF的结合模式与EGFR1阳性染色的模式一致。另外5个肿瘤显示出弱免疫反应性,但在这些肿瘤中未检测到生物素化EGF的结合。其余8个肿瘤在两种技术检测下均为阴性。这些不一致的情况可能反映出配体结合技术的灵敏度较低,或者在这些组织中可能表达了异常受体。用生物素化EGF可在大鼠肝脏中检测到EGF受体,但用抗体则检测不到,这表明基于配体的技术在跨物种研究中的有用性。