目的：以NO供体巯基亚硝基-谷胱甘肽(S-nitrosoglutathione GSNO)作为工具药，用已构建的人鼻咽癌细胞（Human nasopharyngeal carcinoma cell, CNE-2)为模型，筛选并分析经GSNO作用后的细胞与对照组细胞间的差异蛋白。方法：以GSH和NaNO2（1:1）方式合成GSNO并在避光酸性条件下保存，利用分光光度计法定量。将培养的GNE-2细胞实验组加GNSO处理，分别对实验组和对照组细胞抽提蛋白质，测量蛋白浓度。利用iTRAQ试剂盒标记蛋白肽段联合LC-MS/MS技术筛选并鉴定差异蛋白。利用GO功能分析、String构图对差异蛋白进行生物信息学分析。结果：本次实验获得两组的总蛋白有758个，根据筛选条件，我们共筛选出差异蛋白107个，其中53种相对于对照组表达增加(<0。5倍)，54种表达下降(>1。7倍)。利用GO分析发现差异蛋白参与人体的14种生物过程，分别代表了10种分子功能类型和9种细胞成分类型。再利用String软件构建了差异蛋白相互作用网络图。结论：NO会使人鼻咽癌细胞中蛋白表达发生变化，寻找差异性明显的蛋白可能为鼻咽癌细胞潜在的药物靶点，可为进一步研究抑制鼻咽癌细胞增殖的机制提供实验基础。89965

Objective: The NO donor nitroso glutathione thiol (GSNO) as a tool in medicine, human nasopharyngeal carcinoma cell line(CNE-2) has been constructed as a model, to study the effect of GSNO on the proliferation of CNE-2 cells, screen out to analyze differential proteins between CNE-2 after GNSO treatment and the control。 Method: GSNO was synthesized by the reaction of GSH and NaNO2 (1:1) under the condition of acid light avoidance and quantified by spectrophotometer。 Add GNSO in experimental group, and extract proteins from the experimental group and the control and measure the concentration of protein。 Combining the iTRAQ label and LC-MS / MS technology to explore and identify differential proteins。 Play GO function analysis and String analysis。 Results: In this experiment, there are 758 proteins identified in total, 107 of which were significantly different between the two groups。 Among which 53 kinds protein expression was increased after GSNO treatment comparing to the control group(<0。5 fold),54 kinds of expression decreased (>1。7fold)。 These differential proteins involve in 14 types of biological processes and represents 10 types of molecular function and 9 types of cellular component。 Construct differential protein interaction network。 Use String software to construct the protein-protein interaction network。 Conclusion: NO will change the protein expression in nasopharyngeal carcinoma cells, finding significant protein may be potential drug targets for nasopharyngeal carcinoma cells, to provide experimental basis for further study of mechanism for inhibiting the proliferation of nasopharyngeal carcinoma cells。

毕业论文关键词：GSNO； 人鼻咽癌细胞株； 蛋白组学

Keyword: Mercapto-nitroso glutathione；Human nasopharyngeal carcinoma cell line；proteomic

目录

目录 3

1引言 4

2材料与方法 5

2。1实验材料及仪器 5

2。1。1细胞株 5

2。1。2实验试剂 5

2。1。3实验仪器 5

2。2方法 6

2。2。1 CNE-2细胞培养 6

2。2。2 GSNO的合成与定量 6

2。2。3 GSNO处理细胞及源Q于W优E尔A论S文R网wwW.yOueRw.com 原文+QQ75201,8766 总蛋白的提取定量 6

2。2。4 iTRAQ试剂的标记及检测