Suter M M, Augustin-Voss H G, Pantano D M, Flanders J A, Varvayanis M
Dept. of Pathology, New York State College of Veterinary Medicine, Cornell University, Ithaca 14853.
J Histochem Cytochem. 1991 Aug;39(8):1103-12. doi: 10.1177/39.8.1856458.
We used lectins as probes to demonstrate the composition of membrane carbohydrates of canine keratinocytes in various functional stages and various degrees of differentiation. Keratinocytes during normal epidermal turnover were compared by lectin immunohistochemistry to cells of hyperplastic epidermis and neoplastic keratinocytes. Three types of epidermal tumors and oral squamous cell carcinomas were examined. In addition, two in vitro tissue culture systems for keratinocytes were studied and compared with in vivo epithelium. In normal skin, PNA reacted only weakly with basal cells, whereas in hyperplastic skin basal cells bound this lectin strongly, demonstrating increasing expression of PNA binding sites with increasing thickness of the stratified squamous epithelium. ConA bound to basal cell tumors only. In oral squamous cell carcinomas, the expression of distinct lectin binding sites correlated with certain histological growth patterns, e.g., UEA-I reacted with highly invasive tumors but not with tumors showing a solid growth pattern. Using cell surface iodination and polyacrylamide gel electrophoresis, distinct differences in cell membrane protein expression were demonstrated between normal and neoplastic keratinocytes. SDS-polyacrylamide gel electrophoresis of cultured normal and neoplastic keratinocytes revealed several cell surface proteins that are specific for either cell type. Neoplastic cells specifically express a 140 KD lectin binding cell surface glycoprotein. The results of this study show that lectin binding patterns of keratinocytes are dependent on the functional state and the degree of differentiation of the cells and demonstrate correlation of some histological growth patterns with distinct lectin binding phenotypes, suggesting association of expression of cell membrane carbohydrate moieties with growth patterns. In addition, close similarities between "lifted cultures" grown at the air-liquid interface and native tissue demonstrate the value of this culture system as a model for differentiated stratified squamous epithelium.
我们使用凝集素作为探针,以证明处于不同功能阶段和不同分化程度的犬角质形成细胞的膜碳水化合物组成。通过凝集素免疫组织化学,将正常表皮更替过程中的角质形成细胞与增生性表皮细胞和肿瘤性角质形成细胞进行比较。检查了三种类型的表皮肿瘤和口腔鳞状细胞癌。此外,研究了两种角质形成细胞的体外组织培养系统,并与体内上皮组织进行比较。在正常皮肤中,PNA仅与基底细胞发生微弱反应,而在增生性皮肤中,基底细胞与这种凝集素强烈结合,表明随着复层鳞状上皮厚度的增加,PNA结合位点的表达增加。ConA仅与基底细胞瘤结合。在口腔鳞状细胞癌中,不同凝集素结合位点的表达与某些组织学生长模式相关,例如,UEA-I与高侵袭性肿瘤反应,但与呈实体生长模式的肿瘤无反应。通过细胞表面碘化和聚丙烯酰胺凝胶电泳,证明了正常和肿瘤性角质形成细胞之间细胞膜蛋白表达存在明显差异。培养的正常和肿瘤性角质形成细胞的SDS-聚丙烯酰胺凝胶电泳显示了几种特定于每种细胞类型的细胞表面蛋白。肿瘤细胞特异性表达一种140KD的凝集素结合细胞表面糖蛋白。本研究结果表明,角质形成细胞的凝集素结合模式取决于细胞的功能状态和分化程度,并证明了一些组织学生长模式与不同凝集素结合表型之间的相关性,提示细胞膜碳水化合物部分的表达与生长模式相关。此外,在气液界面生长的“悬浮培养物”与天然组织之间的密切相似性证明了这种培养系统作为分化复层鳞状上皮模型的价值。