胃癌细胞周期基因表达谱的变化  

Analysis of gene expression profiles in gastric cancer cell cycle

作　　者：兰斌[1] 刘炳亚[1] 陈雪华[1] 张济 王侃侃 朱正纲[1] 

机构地区：[1]上海第二医科大学附属瑞金医院外科上海消化外科研究所 [2]上海市血液病研究所医学基因组国家重点实验室

出　　处：《中华肿瘤杂志》2006年第8期568-571,共4页Chinese Journal of Oncology

基　　金：国家重点基础研究发展计划(973计划)资助项目(2002CB713700);福建医科大学科学研究发展基金资助项目(FJGXY04027)

摘　　要：目的检测胃癌MKN45细胞周期基因表达谱的变化,从基因组学角度阐明胃癌细胞增殖的机制。方法应用双胸腺嘧啶核苷法和胸腺嘧啶核苷-偌考达唑法,分别阻断胃癌细胞株MKN45于G2／M和G1／S交界点后释放;流式细胞仪监测细胞同步化程度;cDNA基因芯片检测细胞周期中G2／M交界点、M／G1过渡期、G1早期、G1晚期、G1／S交界点、S早期、S晚期、G2早期和G2末期的基因表达谱,专业软件进行聚类分析;借助基因数据库,重点对MKN45细胞周期G1末期及G2期上调基因表达进行分析。定最PCR测定cyclin E、cyclin B、plk1、STK15基因在上述9个时间点的mRNA水平变化。结果分析基因芯片检测,得到9个时间点均可信的2001个基因,其中959个基因出现改变(上调或下调),在G1末期或G2期上调基因379个,S期和M期上调基因40个(与G1末期和G2期上凋基因重合),在G1末期上调基因中,功能主要涉及DNA代谢、转录与翻译、蛋白质转运、泛素化和信号转导等;G2期上调基因主要涉及RNA合成与加工、蛋白质转运、细胞骨架合成、凋亡与抑凋亡、转录调节、泛素化、信号转导、有丝分裂调节以及癌基因的表达等。定量PCR检测4个基因的mRNA水平变化趋势,与基因芯片检测结果基本一致。结论在MKN45细胞周期演进中,DNA复制及染色体分离所需的各种物质准备分别在G,末期及G2期完成,这种准备涉及多种类基因,成为推动MKN45细胞周期进程的主要动力,其中部分基因可能与肿瘤的过度增殖有关。基因芯片检测结果具有较好的可信度,为今后胃癌细胞周期相关基因功能研究提供了帮助。Objective To detect the gene expression profile in gastric cancer cell cycle and explain the mechanism of gastric cancer cell proliferation by a genomic study. Methods Gastric cancer ceils MKN45 were synchronized at G2/M and G1/S point by nocodazole-thymidine and double thymidine methods. The synchronizing degree of cells was monitored by flow cytometry. The gene expression profiles at G2/M point, M/G1 transition, G1 early phase, G1 late phase, G1/S point, S early phase, S late phase, G2 early phase and G2 late phase in MKN45 cell cycling were examined using cDNA microarray chips. Hierarchy analysis was conducted with a professional software package and the up-regulated genes at G1 late and G2 phase were analyzed according to gene database. Furthermore, the mRNA level of cyclin E, cyclin B, plkl and STK15 in above mentioned nine points were measured by quatitative PCR. Results 2001 genes were detected to be available at all 9 points via software processing, out of which 959 appeared up-regulated or down-regulated. 379 genes showed to be up-regulated at late G1 （147） or G2 phases （232） , 40 at S and M phases （also up-regulated at G1 late and G2 phases）. The 147 up-regulated genes at G1 late phase are involved in DNA metabolism, transcription and translation, protein transportation, ubiquitination and signal transduction, etc. The 232 up-regnlated genes in G2 phase are involved in RNA synthesis and processing, intracellular protein transportation, cytoskeleton synthesis, signal transduction, apoptosis and anti-apoptosis, transcription regulation, ubiquitination, mitosis regulation and oncogene expression, etc. The mRNA level of 4 genes detected by quantitative PCR during cell cycle was in agreement with that detected by microarray. Conclusion During MKN45 cell cycling, the preparation for DNA synthesis and chromosome separation are conducted in G1 and G2 , which are implicated in multiple genes, may be the main impetus of driving MKN45 cell cycle. Some of these genes may be related to tumor over-prolife

关 键 词：胃肿瘤 MKN45细胞周期 基因表达谱 

分 类 号：R735.2[医药卫生—肿瘤]