de Wit P E, Moretti S, Koenders P G, Weterman M A, van Muijen G N, Gianotti B, Ruiter D J
Department of Pathology, University Hospital Nijmegen, The Netherlands.
J Invest Dermatol. 1992 Aug;99(2):168-73. doi: 10.1111/1523-1747.ep12616793.
Different results have been reported on the expression of epidermal growth factor receptor (EGFR) in human melanocytic lesions, which may be due to different methodologic approaches. Therefore, we compared EGFR expression in six human melanoma cell lines by utilizing the monoclonal antibodies 2E9, 425, and 225, applying four immunocytochemical staining procedures. The results were compared with those obtained by a multiple point ligand binding assay. In addition, Northern blot analysis was performed. A three-step immunoperoxidase method using the monoclonal antibody 2E9 proved most sensitive. Staining intensities, estimated semiquantitatively, correlated well with the quantitative data obtained by the ligand-binding assay. Expression on the mRNA level was also in agreement with these results. Immunohistochemical staining of a large series of human cutaneous melanocytic lesions using the method selected showed differential EGFR expression in various stages of melanocytic tumor progression: 19% of common nevocellular nevi; 61% of dysplastic nevi, 89% of primary cutaneous melanomas, and 91% of melanoma metastases showed staining of the melanocytic cells. Intralesional heterogeneity of EGFR expression was present. Although the mean percentage of positive melanocytic cells in positive lesions did not increase with progression, mean staining intensity was stronger in malignant lesions compared to benign lesions. Ligand binding assays showed that EGFR expression in the highly metastasizing cell lines MV3 and BLM was at least 40 times higher than in the cell lines IF6, 530, M14, and Mel57, which do not or only sporadically metastasize after subcutaneous inoculation in nude mice. Although the differences between the various stages of progression are not absolute, we provide further evidence that EGFR expression increases in human melanocytic tumor progression.
关于人类黑素细胞性病变中表皮生长因子受体（EGFR）的表达，已有不同的报道结果，这可能是由于采用了不同的方法学途径。因此，我们利用单克隆抗体2E9、425和225，应用四种免疫细胞化学染色程序，比较了六种人类黑色素瘤细胞系中EGFR的表达情况。将结果与通过多点配体结合测定法获得的结果进行了比较。此外，还进行了Northern印迹分析。使用单克隆抗体2E9的三步免疫过氧化物酶法被证明是最敏感的。半定量估计的染色强度与通过配体结合测定获得的定量数据相关性良好。mRNA水平的表达也与这些结果一致。使用所选方法对大量人类皮肤黑素细胞性病变进行免疫组织化学染色，结果显示在黑素细胞肿瘤进展的各个阶段EGFR表达存在差异：19%的普通痣细胞痣；61%的发育异常痣，89%的原发性皮肤黑色素瘤，以及91%的黑色素瘤转移灶显示黑素细胞染色。存在病变内EGFR表达的异质性。尽管阳性病变中阳性黑素细胞的平均百分比并未随进展而增加，但与良性病变相比，恶性病变中的平均染色强度更强。配体结合测定表明，高转移细胞系MV3和BLM中的EGFR表达至少比在皮下接种于裸鼠后不转移或仅偶尔转移的细胞系IF6、530、M14和Mel57中的EGFR表达高40倍。尽管进展的各个阶段之间的差异并非绝对，但我们提供了进一步的证据表明，在人类黑素细胞肿瘤进展过程中EGFR表达增加。
