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[内质网蛋白57基因启动子甲基化与维吾尔族女性宫颈病变发病机制的关联]

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

作者信息

Abdula Mangnishahan, Abudulajiang Gulnaer, Amiduo Reshalaiti, Abudala Abulizi, Hasim Ayshamgul

机构信息

Department of Medical Oncology, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2013 Aug;35(8):600-3.

Abstract

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_1, 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 island-specific, but varies for individual CpG sites, and may explain to a certain extent the epigenetic mechanisms regulated by Erp57 gene expression.

摘要

目的

探讨内质网蛋白57(ERp57)基因启动子区域甲基化与维吾尔族女性宫颈病变发病机制之间的关系及意义。

方法

运用专门软件设计ERp57基因启动子CpG岛片段的特异性引物,对经亚硫酸氢盐修饰的SiHa癌细胞DNA进行PCR扩增,对目标片段进行克隆及测序,以获取基因碱基序列中CpG甲基化的相关信息。收集78例CIN、CSCC新鲜组织及正常对照组织,采用Sequenom MassARRAY(DNA)技术检测不同宫颈病变中ERp57基因启动子区域的甲基化水平。

结果

ERp57基因相应目标片段包含18个CpG位点。所有CpG位点甲基化均出现在SiHa宫颈癌细胞基因组DNA中。通过Sequenom MassARRAY平台对单个CpG位点甲基化进行定量分析所得数据显示,在CSCC、CIN或对照组中,来自CpG_1、CpG_2、CpG_3.4、CpG_5、CpG_6、CpG_7、CpG_8和CpG_9的三个CpG位点(CpG_1、CpG_5和CpG_7)之间的甲基化水平存在显著差异。

结论

尽管维吾尔族女性CSCC中ERp57基因启动子的整体甲基化水平高于CIN和正常对照组织,但高甲基化仅发生在某些CpG岛和位点。这表明DNA甲基化对表达的调控并非CpG岛特异性的,而是因单个CpG位点而异,这在一定程度上可能解释了Erp57基因表达调控的表观遗传机制。

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