维吾尔族妇女宫颈上皮内瘤变和宫颈癌与内质网蛋白57基因启动子甲基化的关系  

Association of promoter methylation of ERp57 gene with the pathogenesis of cervical lesions in Uighur women

作　　者：忙尼沙汗·阿不都拉[1] 古丽那尔·阿布都拉江[2] 热沙来提·艾米多[3] 阿布力孜·阿布杜拉[4] 阿仙姑·哈斯木[3] 

机构地区：[1]新疆医科大学第一附属医院肿瘤科,乌鲁木齐830011 [2]新疆医科大学第一附属医院病理科 [3]新疆医科大学病理教研室 [4]生物化学教研室

出　　处：《中华肿瘤杂志》2013年第8期600-603,共4页Chinese Journal of Oncology

基　　金：国家自然科学基金（81060164）

摘　　要：目的 探讨宫颈上皮内瘤变和宫颈癌与内质网蛋白57（ERp57）基因启动子区甲基化水平的关系和意义。方法应用MethPrimers软件设计ERp57基因启动子区CpG岛特异性PCR引物，对宫颈癌SiHa细胞DNA进行亚硫酸氢盐修饰、目的片段扩增、质粒载体克隆和测序，确定目的片段所含CpG序列甲基化情况。收集15例正常宫颈上皮（对照组）、30例宫颈上皮内瘤变（CIN组）和33例宫颈鳞癌（CSCC组）患者的新鲜组织标本，应用基质辅助激光解吸电离飞行时间质谱（MALDI—TOF）技术定量分析宫颈病变组织DNAERp57基因启动子单-CpG位点的甲基化水平。结果ERp57基因相应目的片段各含9个CpG位点，在宫颈癌SiHa细胞基因组DNA中，所有的CpG位点都发生了甲基化，甲基化率为100％。MALDI—TOF检测结果显示，在CSCC组中，CpG-1、CpG_5和CpG．_7位点的甲基化水平分别为0．9021±0．1810、0．85214-0．1696和0．9671±0．1425，对照组分别为0．7721±0．0644、0．7347±0．0924和0．8765±0．0226，差异均有统计学意义（均P〈0．05）。CSCC组、CIN组和对照组宫颈组织ERp57基因的甲基化水平分别为0．9065±0．3709、0．8697±0．3336和0．8040±0．4705，差异无统计学意义（P=0．052）。结论维吾尔族妇女宫颈病变组织中，ERp57基因启动子某些单个CpG位点发生甲基化后，引起该基因转录下调，其位点可能是关键性CpG位点。Objective To investigate the relationship and significance between endoplasmic reticulum protein 57 （ERp57） gene promoter region methylation with the pathogenesis of cervical lesions in Uighur women. Methods The special software was used to design specific primers of CpG island fragments of ERp57 gene promoter and bisulfite-modified SiHa cancer cell DNA for PCR amplification, cloning and sequencing the target fragments to obtain relevant information of CpG methylation in the gene base sequencs. Seventy-eight fresh tissues of CIN, CSCC and normal control were collected, and the methylation level of ERp57 gene promoter regions in different cervical lesions were identified using Sequenom MassARRAY DNA technology. Results ERp57 gene corresponding target fragment contained the 18 CpG sites. All of the CpG sites methylation occurred in SiHa cervical cancer cell genomic DNA. The analysis of the data resulted from the quantitative analysis of single CpG site methylation by Sequenom MassARRAY platform showed that the methylation level between three CpG sites （ CpG_1, CpG_5 and CpG_7） from CpG_I, CpG_2, CpG_3.4, CpG_5, CpG_6, CpG_7, CpG_8 and CpG_9 had significant differences in the CSCC, CIN or control groups. Conclusions Although the global methylation level of the ERp57 gene promoter is higher in CSCC than that in CIN and normal control tissues in Uighur women, hypermethylation occurs only in certain CpG islands and sites. This indicates that the regulation of expression by DNA methylation is not CpG islandspecific, but varies for individual CpG sites, and may explain to a certain extent the epigenetic mechanisms regulated by Erp57 gene expression.

关 键 词：宫颈肿瘤 宫颈上皮内瘤样病变 DNA甲基化 内质网蛋白57 维吾尔族 

分 类 号：R737.33[医药卫生—肿瘤]