Young A R, Potten C S, Nikaido O, Parsons P G, Boenders J, Ramsden J M, Chadwick C A
Department of Photobiology, St John's Institute for Dermatology, St Thomas Hospital, London, UK.
J Invest Dermatol. 1998 Dec;111(6):936-40. doi: 10.1046/j.1523-1747.1998.00435.x.
Epidemiology shows a relationship between solar exposure and all types of skin cancer. Understanding the mechanisms of skin cancer requires knowledge of the photomolecular events that occur within the relevant epidermal cell types in vivo. Studies to date have focused on UVR-induced DNA lesions in keratinocytes, the majority epidermal cell population which gives rise to most skin cancers. Malignant melanoma, arising from melanocytes (5%-10% of epidermal cells), accounts for most skin cancer deaths. We report on new techniques to detect DNA photolesions in human epidermal melanocytes in situ. Previously nonexposed buttock skin of volunteers of skin types I/II was exposed to clinically relevant doses of narrow bandwidth UVB (300 nm) and UVA (320 nm, 340 nm, 360 nm) radiation. Biopsies were taken immediately afterwards and processed for routine histology. Microscope sections were prepared and double-stained with fluorescent-tagged monoclonal antibodies for thymine dimers and melanocytes. UVR dose-response curves for dimer levels within melanocyte nuclei were determined by image analysis and compared with dimer levels in adjacent basal cell keratinocytes. Our data show that UVB and UVA readily induce thymine dimers in melanocytes at levels that are comparable with those found in adjacent keratinocytes. This new technique will enable melanocyte specific studies, such as DNA repair kinetics, to be done in vivo.
流行病学研究表明,日光照射与所有类型的皮肤癌之间存在关联。要了解皮肤癌的发病机制,需要知晓体内相关表皮细胞类型中发生的光分子事件。迄今为止的研究主要聚焦于紫外线辐射(UVR)诱导角质形成细胞中的DNA损伤,角质形成细胞是表皮中的主要细胞群体,大多数皮肤癌都由其引发。恶性黑色素瘤起源于黑素细胞(占表皮细胞的5%-10%),却是导致大多数皮肤癌死亡的原因。我们报告了一种原位检测人表皮黑素细胞中DNA光损伤的新技术。将I/II型皮肤志愿者先前未暴露的臀部皮肤暴露于临床相关剂量的窄带宽UVB(300纳米)和UVA(320纳米、340纳米、360纳米)辐射下。之后立即进行活检并进行常规组织学处理。制备显微镜切片,并用荧光标记的单克隆抗体对胸腺嘧啶二聚体和黑素细胞进行双重染色。通过图像分析确定黑素细胞核内二聚体水平的UVR剂量反应曲线,并与相邻基底细胞角质形成细胞中的二聚体水平进行比较。我们的数据表明,UVB和UVA很容易在黑素细胞中诱导胸腺嘧啶二聚体,其水平与在相邻角质形成细胞中发现的水平相当。这项新技术将能够在体内开展黑素细胞特异性研究,如DNA修复动力学研究。