中波紫外线与化学致癌物诱导下HaCaT细胞的蛋白表达应答  

Protein Expression Alternations for the HaCaT Proteome in Response to the UVB-irradiation and Chemical-carcinogen

作　　者：孙明忠[1] 刘淑清[2] 吕晓[2] 郭一萌[1,2] 郭春梅[1,2] 辛毅[1] 唐建武[3] 

机构地区：[1]大连医科大学基础医学院生物技术系,大连116044 [2]大连医科大学生物化学与分子生物学教研室,大连116044 [3]大连医科大学病理学教研室,大连116044

出　　处：《高等学校化学学报》2009年第10期1965-1971,共7页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30572098);大连医科大学科研启动经费(批准号:2007001;2007003)资助

摘　　要：对角质形成细胞HaCaT分别进行中波紫外线(UVB)照射、2,4-二硝基苯磺酸(DNBS)刺激及UVB+DNBS(UD)共同刺激,利用二维荧光差异胶内凝胶电泳(2D DIGE)、DeCyder定量分析软件和HPLC-nESI-MS/MS分析技术,对HaCaT细胞产生的差异表达蛋白进行了鉴定.有65个蛋白质点发生了明显表达差异(P<0.05),与UVB或DNBS单独处理细胞相比,有41个蛋白点表现为UVB和DNBS的正协同效应,13个蛋白点表现为负协同效应,5个蛋白点与UVB单独处理相近,6个蛋白点与DNBS单独处理相近.HPLC-nESI-MS/MS从65个差异表达蛋白质点中共鉴定出60种单一(Unique)蛋白.采用生物信息学方法对这些鉴定蛋白所涉及的分子功能、参与的生物学过程及信号通路进行了系统分析.实验得到了与紫外辐射和化学诱导损伤的直接相关蛋白,有助于研究不同环境条件下皮肤癌的形成及皮肤疾病的有效防护与治疗.The keratinoeyte cell line, HaCaT, was radiated by UVB-irradiation, induced by the 2,4-dinitrobenzene sulfonic acid （DNBS） and stimulated by the combination of UVB-irradiation and DNBS-stimulation （UD） , respectively. The differentially expressed protein candidates in response to those stimulations were separated by two-dimensional fluorescent in gel electrophoresis （2D DIGE ）, and the expression levels were quantified by DeCyder software and identifications of interested proteins were performed by the high performance liquid chromatography nano eleetrospray tandem mass spectrometry （HPLC-nESI-MS/MS） analysis. Sixty-five protein spots were found differentially expressed with metastatic significance （P 〈 0. 05 ）. Comparing to the HaCaT cell treated either by UVB alone or DNBS alone, forty-one and thirteen protein spots were shown positively cooperatively regulated and negatively cooperatively regulated by the stimulations of UVB plus DNBS, respectively ; and there are five and six protein spots show similar expression levels as the cells being treated with UVB alone and stimulated with DNBS alone. Sixty unique proteins were identified from all the sixtyfive protein spots by HPLC-nESI-MS/MS analysis. Those protein candidates were systemically categorized by bioinformatics analysis based on their molecular functions, involvements in biological processes and pathways. The results from current study provide the protein candidates directly associated with the UVB irradiation and chemical stimualtion, which helps to investigate the formation mechanisms of skin cancers under different environmental conditions and to prevent and treat skin diseases.

关 键 词：HaCaT细胞系 中波紫外线 2 4-二硝基苯磺酸 二维荧光差异胶内凝胶电泳 

分 类 号：O629[理学—有机化学]