Gould V E, Koukoulis G K, Jansson D S, Nagle R B, Franke W W, Moll R
Department of Pathology, Rush Medical College, Chicago, Illinois 60612.
Am J Pathol. 1990 Nov;137(5):1143-55.
The authors studied by immunohistochemistry the intermediate filament (IF) protein profile of 66 frozen samples of breast tissue, including normal parenchyma, all variants of fibrocystic disease (FCD), fibroadenomas, cystosarcoma phylloides, and ductal and lobular carcinomas. Monoclonal antibodies (MAbs) to cytokeratins included MAb KA 1, which binds to polypeptide 5 in a complex with polypeptide 14 and recognizes preferentially myoepithelial cells; MAb KA4, which binds to polypeptides 14, 15, 16 and 19; individual MAbs to polypeptides 7, 13, and 16, 17, 18, and 19, and the MAb mixture AE1/AE3. The authors also applied three MAbs to vimentin (Vim), and three MAbs to glial filament protein (GFP). Selected samples were studied by double-label immunofluorescence microscopy and by staining sequential sections with some of the said MAbs, an MAb to alpha-smooth muscle actin, and well-characterized polyclonal antibodies for the possible coexpression of diverse types of cytoskeletal proteins. Gel electrophoresis and immunoblot analysis also were performed. All samples reacted for cytokeratins with MAbs AE1/AE3, although the reaction did not involve all cells. Monoclonal antibody KA4 stained preferentially the luminal-secretory cells in the normal breast and in FCD, whereas it stained the vast majority of cells in all carcinomas. Monoclonal antibody KA1 stained preferentially the basal-myoepithelial cells of the normal breast and FCD while staining tumor cell subpopulations in 4 of 31 carcinomas. Vimentin-positive cells were found in 8 of 12 normal breasts and in 12 of 20 FCD; in most cases, Vim-reactive cells appeared to be myoepithelial, but occasional luminal cells were also stained. Variable subpopulations of Vim-positive cells were noted in 9 of 20 ductal and in 1 of 7 lobular carcinomas. Glial filament protein-reactive cells were found in normal breast lobules and ducts and in 15 of 20 cases of FCD; with rare exceptions, GFP-reactivity was restricted to basally located, myoepithelial-appearing cells. Occasional GFP-reactive cells were found in 3 of 31 carcinomas. Evaluation of sequential sections and double-label immunofluorescence microscopy showed the coexpression of certain cytokeratins (possibly including polypeptides 14 and 17) with vimentin and alpha-smooth muscle actin together with GFP in some myoepithelial cells. The presence of GFP in myoepithelial cells was confirmed by gel electrophoresis and immunoblotting. Our results indicate that coexpression of cytokeratin with vimentin and/or GFP is comparatively frequent in normal basal-myoepithelial cells of the breast.(ABSTRACT TRUNCATED AT 400 WORDS)
作者采用免疫组织化学方法研究了66份乳腺组织冷冻样本的中间丝（IF）蛋白谱，这些样本包括正常实质组织、纤维囊性疾病（FCD）的所有变体、纤维腺瘤、叶状囊肉瘤以及导管癌和小叶癌。针对细胞角蛋白的单克隆抗体（MAb）包括与多肽5结合形成复合物并优先识别肌上皮细胞的MAb KA 1；与多肽14、15、16和19结合的MAb KA4；针对多肽7、13以及16、17、18和19的单个MAb，以及MAb混合物AE1/AE3。作者还应用了三种针对波形蛋白（Vim）的MAb和三种针对神经胶质丝蛋白（GFP）的MAb。通过双标记免疫荧光显微镜以及用上述部分MAb、一种针对α-平滑肌肌动蛋白的MAb和特性明确的多克隆抗体对连续切片进行染色，研究选定样本中不同类型细胞骨架蛋白的可能共表达情况。还进行了凝胶电泳和免疫印迹分析。所有样本与MAb AE1/AE3反应显示细胞角蛋白阳性，尽管并非所有细胞都有反应。单克隆抗体KA4优先染色正常乳腺和FCD中的腔分泌细胞，而在所有癌中它染色绝大多数细胞。单克隆抗体KA1优先染色正常乳腺和FCD的基底肌上皮细胞，而在31例癌中的4例中染色肿瘤细胞亚群。在12例正常乳腺中的8例以及20例FCD中的12例中发现波形蛋白阳性细胞；在大多数情况下，Vim反应性细胞似乎是肌上皮细胞，但偶尔也有腔细胞被染色。在20例导管癌中的9例以及7例小叶癌中的1例中注意到Vim阳性细胞的不同亚群。在正常乳腺小叶和导管以及20例FCD中的15例中发现神经胶质丝蛋白反应性细胞；除罕见例外，GFP反应性仅限于位于基底的、呈肌上皮样的细胞。在31例癌中的3例中偶尔发现GFP反应性细胞。对连续切片和双标记免疫荧光显微镜的评估显示，在一些肌上皮细胞中某些细胞角蛋白（可能包括多肽14和17）与波形蛋白、α-平滑肌肌动蛋白以及GFP共表达。通过凝胶电泳和免疫印迹证实了肌上皮细胞中存在GFP。我们的结果表明，在乳腺正常基底肌上皮细胞中细胞角蛋白与波形蛋白和/或GFP的共表达相对常见。（摘要截选至400字）
