Bäckvall Helena, Wassberg Cecilia, Berne Berit, Ponten Fredrik
Department of Genetics and Pathology, University Hospital, S-751 85 Uppsala, Sweden.
Exp Dermatol. 2002 Aug;11(4):349-56. doi: 10.1034/j.1600-0625.2002.110409.x.
Ultraviolet radiation (UVR) plays an important role in the development of non-melanoma skin cancer. Most tumors develop in chronically sun-exposed skin, most often in cosmetically sensitive locations, where in vivo experiments may be difficult to perform. In this study, we describe a skin organ culture model with preserved normal morphology and intact response to UVR. Skin explants from chronically sun-exposed and non-sun-exposed skin were irradiated with artificial UVA+UVB with and without topical sunscreen. UV-induced DNA damage, epidermal p53 response and repair kinetics were analyzed using immunohistochemistry. Four hours after UV-irradiation epidermal keratinocytes showed a strong immunoreactivity for thymine-dimers. Gradual repair during an incubation time resulted in few residual thymine-dimers after 48 h. Repair appeared to be more efficient in chronically sun-exposed skin compared with non-sun-exposed skin. There was also an accumulation of p53 protein in epidermal keratinocytes, peaking at 4-24 h after irradiation. Large interindividual differences with respect to formation and repair of thymine-dimers as well as induction and duration of the p53 response were observed. Skin explants treated with topical sunscreen prior to UV-irradiation showed a clear reduction of thymine-dimers and p53 expression. The epidermal UV-responses and repair kinetics in organ-cultured skin were similar to what was found in vivo. Our data suggest that organ-cultured skin provides a valuable tool for studies of UV-induced epidermal responses in chronically sun-exposed skin.
紫外线辐射(UVR)在非黑色素瘤皮肤癌的发生发展中起着重要作用。大多数肿瘤发生在长期暴露于阳光下的皮肤,最常见于美容敏感部位,在这些部位进行体内实验可能很困难。在本研究中,我们描述了一种皮肤器官培养模型,其保留了正常形态并对UVR具有完整反应。用人工UVA + UVB照射来自长期暴露于阳光下和未暴露于阳光下皮肤的皮肤外植体,并使用或不使用局部防晒霜。使用免疫组织化学分析紫外线诱导的DNA损伤、表皮p53反应和修复动力学。紫外线照射后4小时,表皮角质形成细胞对胸腺嘧啶二聚体显示出强烈的免疫反应性。在孵育期间的逐渐修复导致48小时后残留的胸腺嘧啶二聚体很少。与未暴露于阳光下的皮肤相比,长期暴露于阳光下的皮肤修复似乎更有效。表皮角质形成细胞中也有p53蛋白的积累,在照射后4 - 24小时达到峰值。观察到胸腺嘧啶二聚体的形成和修复以及p53反应的诱导和持续时间存在很大的个体差异。在紫外线照射前用局部防晒霜处理的皮肤外植体显示胸腺嘧啶二聚体和p53表达明显减少。器官培养皮肤中的表皮紫外线反应和修复动力学与体内发现的相似。我们的数据表明,器官培养皮肤为研究长期暴露于阳光下的皮肤中紫外线诱导的表皮反应提供了一个有价值的工具。