摘要
目的:初步探讨紫外线辐射所致皮肤细胞骨架蛋白质组整体变化规律。方法:分别提取30 mJ/cm^2中波紫外线(UVB)照射前后角质形成细胞HaCaT株和10 J/cm^2长波紫外线(UVA)照射前后成纤维细胞的总蛋白,采用固相pH梯度双向凝胶电泳技术进行分离,经Imaging Master 2D软件分析双向电泳图谱以发现差异表达蛋白,并对部分差异表达蛋白进行胶内酶切、基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析,测定其肽质量指纹图谱,在网上蛋白质数据库检索鉴定差异蛋白。结果:获得良好的双向电泳图谱,图像分析显示在紫外线辐射前后很多蛋白质发生差异表达,对差异表达的部分蛋白点进行肽质量指纹图谱分析,经Mascot软件检索人非冗余蛋白质数据库后,鉴定出7个涉及皮肤细胞骨架的蛋白质:微管蛋白α2和微管蛋白β5,角蛋白K7、角蛋白K8、角蛋白K13、角蛋白K18和角蛋白K19。对它们的功能和与紫外线辐射的可能关系进行了初步探讨。结论:紫外线辐射可以诱导皮肤细胞骨架蛋白质组发生改变,而这些骨架蛋白的变化不仅与皮肤细胞应对紫外线辐射并清除其损伤密切相关,还可能进一步参与紫外线诱导的细胞凋亡及细胞癌变。
Objective： To reveal the proteomic change of cytoskeleton in skin cell induced by UV radiation. Methods： The cultured HaCaT （immobilized human keratinocyte）cells and the fibroblast were irradiated by 30 mJ/cm^2 UVB and 10 J/cm^2 UVA respectively. The total proteins of these skin cells before and after UV radiation were separated by immobilized pH gradient based two dimensional gel electrophoresis （2-DE）. The differential expression proteins were analyzed by using Imaging Master 2D analysis ware. The differential spots were identified by peptide mass fingerprinting based on Matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS） and database searching were performed on the internet. Results： The 2-D image was stable and showed lots of differential protein spots in skin cells after UV radiation. Seven proteins associated with cell skeleton were identified by MALDI-TOF-MS, they were tubulin α2, tubulin β5, keratin 7, Keratin 8, keratin13, keratin18 and keratin19. Conclusion： UV radiation could induce changes of the cytoskeleton protein expression pattern in keratinocyte and fibroblast. This proteomic changes could help skin cells to removed the impairment of UV radiation, and further may induce cell apoptosis or carcinogenesis.
出处
《临床皮肤科杂志》
CAS
CSCD
北大核心
2009年第3期141-144,共4页
Journal of Clinical Dermatology
