Varani J, Trinh D, Carey T E, Liebert M, Wheelock M J
Department of Pathology, University of Michigan Medical School, Ann Arbor, USA.
Invasion Metastasis. 1995;15(5-6):189-96.
Human neonatal foreskin was maintained in organ culture under serum-free, growth-factor-free conditions or in the presence of a combination of growth factors that are known to stimulate keratinocyte proliferation in monolayer culture. Previously, we have shown that normal histology is maintained when growth-factor-free conditions are used but that the epithelium undergoes a hyperproliferative response and invades the dermis in the presence of the exogenous growth factors. In the present study, the tissue was examined by immunofluorescence for expression of alpha 6 and beta 4 integrin components and for E-cadherin. Under growth factor-free conditions, both alpha 6 and beta 4 were localized to the basal surface of epithelial cells in contact with the basement membrane. In contrast, both epitopes were diffusely distributed throughout the basal epithelium in the presence of growth factors. E-cadherin expression was rapidly lost from the tissue in organ culture. This occurred in both the presence and absence of exogenous growth factors. On the basis of these immunochemical results, we conclude that the same changes in alpha 6 and beta 4 expression that are seen in rapidly proliferating keratinocytes and squamous epithelial cell tumors can be seen in the epidermis of organ-cultured skin when it is maintained in the presence of epithelial growth factors. The observed loss of E-cadherin, in contrast, appears to be a consequence of incubation in organ culture.
人新生儿包皮在无血清、无生长因子的条件下或在已知能刺激角质形成细胞在单层培养中增殖的生长因子组合存在的情况下进行器官培养。此前,我们已经表明,使用无生长因子条件时可维持正常组织学,但在外源生长因子存在的情况下,上皮会发生过度增殖反应并侵入真皮。在本研究中,通过免疫荧光检查组织中α6和β4整合素成分以及E-钙黏蛋白的表达。在无生长因子条件下,α6和β4均定位于与基底膜接触的上皮细胞的基底表面。相反,在生长因子存在的情况下,这两个表位均在整个基底上皮中呈弥漫性分布。在器官培养中,组织中的E-钙黏蛋白表达迅速丧失。无论有无外源生长因子,这种情况都会发生。基于这些免疫化学结果,我们得出结论,在器官培养的皮肤表皮中,当在表皮生长因子存在的情况下培养时,α6和β4表达的相同变化也可见于快速增殖的角质形成细胞和鳞状上皮细胞瘤中。相比之下,观察到的E-钙黏蛋白丧失似乎是器官培养孵育的结果。