低浓度MNNG诱发FL细胞应答反应的蛋白质组学研究  

Proteomic analysis of cellular responses to low concentration of N-methyl-N'-nitro-N- nitrosoguanidine in human amnion FL cells

作　　者：金静华[1] 高志华[1] 杨军[1] 余应年[1] 

机构地区：[1]浙江大学医学院病理生理教研室,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2003年第5期375-379,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：国家重点基础研究发展规划 (973)资助(2 0 0 2 CB5 12 90 1) ;国家自然科学基金重点资助 (39830 2 10 )

摘　　要：目的 :研究低浓度甲基硝基亚硝基胍 (MNNG)对人羊膜 FL细胞蛋白质表达谱的影响 ,以助于阐明环境致癌物引起细胞早期应答反应的分子机制。方法 :FL细胞分别用 MNNG和 DMSO(溶剂对照 )处理后 ,提取细胞总蛋白 ,用双向凝胶电泳分离蛋白质组成分 ,银染染色后用相应的分析软件分析数字化的凝胶图像 ,找出在 MN-NG处理后表达有差异的蛋白斑点 ,用基质辅助的激光解吸 -飞行时间质谱 (MAL DI- TOF)鉴定。结果 :在 MNNG处理后有 6 0多个蛋白斑点出现质和量的变化 ,其中 18个蛋白斑点只能在 MNNG处理的细胞中检测到 ,13个蛋白斑点则在 MNNG处理后消失 ;同时检测到 30个蛋白斑点在表达量上有显著变化 ,其中 16个点表达升高 ,14个点则表达降低。通过数据库的检索 ,初步的鉴定了一批结构和功能各异的蛋白。结论 :在低浓度烷化剂攻击后 ,FL细胞中的蛋白表达谱发生了显著的变化。Objective: To investigat the protein profile after treatment of low concentration of N-methyl-N'- nitro-N-nitrosoguanidine (MNNG) in human FL cells. Methods: After MNNG treatment, whole cellular proteins were separated using two-dimensional gel electrophoresis and visualized by silver staining; the digitized images were analyzed with 2D analysis software. The differentially expressed protein spots were identified by matrix- assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). Results: More than 60 proteins showed significant changes in MNNG-treated cells compared to control cells (DMSO treatment). There were 18 protein spots detected only after MNNG treatment, while 13 protein spots were detected only in the control cells. Moreover, the levels of another 31 proteins were either increased or decreased in MNNG-treated FL cells. And some of the proteins were identified by MALDI-TOF-MS. Conclusion: There are significant alterations of protein profile after MNNG attack.

关 键 词：FL细胞/遗传学 N-甲基-N'-硝基-N-亚硝基胍 蛋白质组学 凝胶 电泳 双向 

分 类 号：R363[医药卫生—病理学]