Boland C R, Roberts J A
Veterans Administration Medical Center, San Francisco, California.
J Histochem Cytochem. 1988 Oct;36(10):1305-7. doi: 10.1177/36.10.3138307.
We have developed a novel method for quantitation of lectin binding sites in mucins derived from colon tissues. Binding of peanut agglutinin and wheat germ agglutinin was measured in extracts from normal and malignant human colon epithelium. Binding of wheat germ agglutinin was used as an estimate of the total mucin present in the tissue extract. Peanut agglutinin was found to bind to mucin from normal colon, but at levels that may be difficult to appreciate by fluorescence microscopy. The yield of mucin extracted from colon cancer was more variable than that from normal colon, and the binding ratio of peanut agglutinin to wheat germ agglutinin was greater in extracts from tumors than in normal tissues. Our findings confirm the histological observation that peanut agglutinin binds more avidly to mucins from colon cancer than to those from normal colon. The finding of peanut agglutinin binding sites in mucins front normal colon was not expected. The quantitative technique may have detected small numbers of binding sites not readily appreciable by fluorescence microscopy. Alternatively, the chromatographic method for measuring lectin binding may be sufficiently sensitive to detect nonspecific binding of the lectin to terminal galactose residues other than the Thomsen-Friedenreich antigen.
我们开发了一种新方法，用于定量分析源自结肠组织的粘蛋白中的凝集素结合位点。在正常和恶性人结肠上皮提取物中测量了花生凝集素和麦胚凝集素的结合情况。麦胚凝集素的结合用于估计组织提取物中存在的总粘蛋白。发现花生凝集素可与正常结肠的粘蛋白结合，但通过荧光显微镜观察可能难以察觉其结合水平。从结肠癌中提取的粘蛋白产量比从正常结肠中提取的更具变异性，并且肿瘤提取物中花生凝集素与麦胚凝集素的结合比率高于正常组织。我们的研究结果证实了组织学观察结果，即花生凝集素与结肠癌粘蛋白的结合比与正常结肠粘蛋白的结合更紧密。在正常结肠粘蛋白中发现花生凝集素结合位点是出乎意料的。这种定量技术可能检测到了荧光显微镜不易察觉的少量结合位点。或者，用于测量凝集素结合的色谱方法可能足够灵敏，能够检测到凝集素与除了Thomsen-Friedenreich抗原之外的末端半乳糖残基的非特异性结合。
