【摘要】 目的:研究胃癌细胞全基因组蛋白H3K27的三甲基化水平。方法:收集我科2007年10月~2008年1月间8例胃癌患者的病灶组织和正常胃黏膜组织,采用染色质免疫沉淀联合芯片技术(ChIP-chip)在全基因组范围内对两种组织细胞的组蛋白H3K27进行高通量的检测。随后采用染色质免疫沉淀-实时定量聚合酶联反应(ChIP-qPCR)验证芯片结果,定量反转录聚合酶联反应(qRT-PCR)检测H3K27显著差异基因的mRNA表达水平。用统计软件进行基因筛选。结果:经两种组织细胞组蛋白H3K27三甲基化水平对比,筛选出234个基因存在H3K27显著差异,肿瘤细胞中有71个基因显示有H3K27三甲基化程度增高,161个基因H3K27甲基化程度降低;ChIP-qPCR验证结果与CpG岛芯片检测结果一致。结论:和正常胃黏膜组织细胞相比,胃癌细胞多个基因组蛋白H3K27三甲基化存在显著改变。ChIP-chip检测技术有利于进一步揭示胃癌发生的分子机制,发现新的基因治疗靶点。更多还原

【Abstract】 Objective:To analyze histone H3 lysine 27 trimethylation of genome-wide by ChIP-chip in gastric cancer cell.Methods:Eight cases including tumor tissue and gastric mucosa tissue in gastric cancer patients were collected from Oct.2007,to Jan.2008.For the first time chromatin immunoprecipitation linked to microarrays(ChIP-chip)was adopted to profile the variations in H3K27me3 in CpG island regions in tumor tissue and gastric mucosa tissue.ChIP-qPCR was used to validate the microrray results.Expression analysis by qRT-PCR was performed to confirm correlations between H3K27me3 and gene expression.Results:Two hundred and thirty-four(71 increased and 161decreased) gene displayed significant H3K27me3 differences were found in tumor cell compared with gastric mucosa tissue.The results of ChIP-qPCR were coincided well with microarray.Conclusion:There are significant differences in H3K27me3 profiling between tumor tissue and gastric mucosa tissue,these novel candidate genes may be become potential biomarkers or future therapeutic targets.The ChIP-chip technology will help further reveal gastric cancer molecular mechanisms and discover new therapeutic targets.更多还原