中文摘要：

背景与目的：LCRG1基因是我室利用mRNA差异显示技术克隆的一个喉癌相关抑癌基因。先前的研究显示将LCRG1转染喉癌细胞系Hep-2，发现其具有抑制细胞增殖、软琼脂集落形成及裸鼠成瘤能力的作用。本研究旨在利用比较蛋白质组学的方法筛选LCRG1抑瘤作用相关的蛋白质。方法：抽提稳定高表达LCRG1基因的Hep-2／LCRG1和载体对照组Hep-2／pcDNA3．1（＋）的细胞总蛋白，采用双向凝胶电泳技术分别分离其总蛋白。考马斯亮蓝染色显色，PDQuest图像分析系统分析获取的2-D胶图像，从胶中选取差异表达的蛋白质点．进行基质辅助激光解吸电离飞行时间质谱或电喷雾-四极杆-飞行时间串联质谱分析．最后Mascot软件搜索数据库初步鉴定蛋白质。结果：获得了重复性和分辨率较好的Hep-2／LCRG1和Hep-2／pcDNA3．1（＋）细胞系的双向凝胶电泳图谱．总蛋白质点数分别为1075±43和1027±23，平均匹配率为91％。图像分析差异蛋白质点，质谱鉴定了20个差异表达的蛋白质点，其中MALDI—TOF-MS鉴定了16个，ESI—Q-TOFMS／MS鉴定了4个。对这些初步鉴定的差异蛋白质进行功能分析．发现一些与细胞代谢、转录调控相关的蛋白质。结论：建立了稳定高表达LCRG1基因的Hep-2／LCRG1和载体对照组Hep．2／pcDNA3．1（＋）细胞系的双向凝胶电泳图谱．应用质谱技术识别并鉴定出一些与LCRG1抑瘤作用相关的差异表达蛋白质．为进一步研究LCRG1作用的分子机制提供新的线索。

英文摘要：

BACKGROUND ＆ OBJECTIVE ： Laryngeal carcinoma-related gene 1 （LCRG1）, a novel laryngeal carcinoma-related tumor suppressor gene, was cloned with mRNA differential display method. Previous researches showed LCRG1 might inhibit cell growth, proliferation, colony formation in soft agar, and tumorigenesis of laryngeal carcinoma cell line Hep-2. This study was to screen the proteins associated with the tumor suppressive function of LCRG1 by comparative proteomics method. METHODS： The whole cellular proteins of Hep-2/LCRG1 and Hep-2/ pcDNA3.1 （＋） cells were extracted and seperated by two-dimensional gel electrophoresis （2-DE） technology. After electrophoresis, the gels were stained by Coomassie Brilliant Blue G-250, and analyzed using PDQuest software. The differentially expressed proteins were cut from the gels, analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）, or electrospray ionization-quadrupole time- of-flight MS/MS （ESI-Q-TOF MS/MS）, and identified through searching database with Mascot software. RESULTS_. The well-resolved, reproducible 2-DE patterns of Hep-2/LCRG1 and Hep-2/pcDNA3.1 （＋） cells were established. The total protein spots were 1 075±43 in Hep-2/LCRG1 cells and 1 027＋23 in Hep-2/pcDNA3.1（＋） cells, with an average matching rate of 91%, Using mass spectrometry technology, 20 differential protein spots between the 2 cell lines were identified. Among them, 16 were identified by MALDI-TOF-MS, and 4 were identified by ESI-Q-TOF MS/MS. Some of the identified proteins were characterized as members of cellular transcription and metabolism enzymes. CONCLUSIONS. In this study, 2-DE gels of Hep- 2/LCRG1 cell line with high expression of LCRG1 mRNA and vector control Hep-2/pcDNA3.1 （＋） cell line were established; some differential proteins related to LCRG1 were identified by mass spectrometry. These data will help to illustrate the molecular mechanism of LCRG1.