表达谱芯片对EB病毒诱导甲基化沉默基因的筛选  

METHYLATION SILENCE EBV-INDUCED GENES: A GENOME WIDE SURVEY BY GENE EXPRESSION MICROARRAY

作　　者：王菁[1] 刘霞[1] 韩美艳[2] 吴国才[1] 王云[1] 罗兵[1] 

机构地区：[1]青岛大学医学院微生物学教研室,山东青岛266021 [2]青岛市妇女儿童医疗保健中心遗传科

出　　处：《青岛大学医学院学报》2012年第4期291-294,297,共5页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省科技厅资助项目(ZR2011CM016)

摘　　要：目的筛选EB病毒(EBV)相关胃癌中大量EBV诱导的甲基化沉默基因,为EBV相关肿瘤的研究提供理论依据。方法采用Agilent人类全基因组表达谱(4×44K)芯片,检测去甲基化药物Aza作用前后的EBV阳性和阴性胃癌细胞株,筛选出EBV相关胃癌特有的甲基化沉默基因。应用实时荧光定量PCR和亚硫酸盐测序(BGS)方法对结果进行验证。结果去甲基化药物作用前后的EBV阳性细胞系差异基因与EBV阳性与阴性胃癌细胞系差异基因的共同基因共68条,此为EBV相关胃癌甲基化沉默特异性基因。在差异倍数为1.5和2.0时,分别检测到19和3条基因。实时荧光定量PCR和BGS验证结果显示,筛选基因在EBV阳性胃癌细胞系中处于低表达、高甲基化状态。结论筛选和鉴定EBV阳性细胞株甲基化沉默的抑癌基因,对明确EBV相关肿瘤的发病机制具有重要作用。Objective To screen EBV-induced silence genes in EBV-associated gastric carcinoma and find the mechanism of EBV tumor genesis. Methods By employing Agilent Human Whole Genome Expression Profiling （4 X 44 K） chips, EBV- positive and -negative gastric carcinoma cell lines, before and after treatment of Aza, were detected. The results were validated by real-time fluorescent quantitation PCR and BGS. Results There were 68 common genes in EBV positive cell lines before versus after Aza treatment with EBV＋ versus EBV- cell lines, which were EBV-related methylation silence specific genes of gastric can- cer. At the fold change of 1.5 and 2.0, 19 and 3 genes were detected, respectively. The PCR and BGS revealed that the genes screened in EBV-positive gastric cancer cells were lowly expressed, and highly methylated. Conclusion The selection and identi- fication of silent anti-oncogene in EBV-positive gene rnethylation is of importance in identifying the oathogenesis of tumors.

关 键 词：疱疹病毒4型 人 甲基化 基因 肿瘤抑制 寡核苷酸序列分析 

分 类 号：R735.2[医药卫生—肿瘤]