摘要
目的:应用蛋白质组技术比较人表皮细胞与角膜上皮细胞之间的蛋白质表达差异。方法:实验于2005-11/2006-10在中山大学中山眼科中心国家眼科学重点实验室完成。分别培养人表皮细胞与角膜上皮细胞,裂解原代细胞抽提总蛋白,精确定量后进行双向电泳,扫描电泳凝胶获得二维总蛋白图谱。图谱使用软件辅助比较分析,找出差异表达点,从凝胶中抠取差异点,胶内酶解后用基质辅助激光解吸附/电离飞行时间串级质谱法鉴定差异蛋白。结果:人表皮细胞与角膜上皮细胞蛋白质凝胶分析获得600余个清晰的蛋白质斑点,在比较分析出的26个差异点中初步鉴定出4个差异表达蛋白,分别是细胞内氯离子通道1、Psoriasin、乳酸脱氢酶B和丝氨酸蛋白酶抑制因子。结论:人表皮细胞与角膜上皮细胞在蛋白质表达上有较高的相似性以及较好的可比性,为探索表皮细胞横向分化成角膜上皮细胞的分子机制做出了有益的尝试,并提供了具有一定可操作性的实验方法。
AIM：To compare the protein expression difference of human epidermal cells and corneal epithelial cells by proteomics technique. METHODS： The experiment was performed at the Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University from November 2005 to October 2006. Human epidermal cells and corneal epithelial cells were cultured, respectively. Total protein was extracted after spliting primary cells. Whole cellular protein was presented by two dimensional （2D） electrophoresis. 2D total protein atlas was obtained by scanning electrophoresis gel, and then analyzed with atlas software to find differential expression that was deducted from gel. After in-gel protein digestion, the different-expressed spots were detected by matrix assisted laser desorption ionization-time of flight mass spectrometry （MALDI-TOF-MS）. RESULTS：About 600 protein spots from the cells were gained after human epidermal cells and corneal epithelial cells gel analysis. A total of 26 proteins with changed expression level were found, and four different-expressed proteins were detected. There were chloride intracellular channel 1 and Psoriasin and Human E, Lactate Dehydrogenase and serine proteinase inhibitor. CONCLUSION： The protein expressions of human epidermal cells and corneal epithelial cells are considerable similarity and comparability. These proteins may provide operational research targets for molecular mechanisms of potential of epidermal cells trans-differentiation into corneal epithelium-like cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第28期5531-5535,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金委员会专项基金/主任基金(No.30640047)
广东省科学事业/科研条件计划项目(No.2006B60101058)~~
