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目的通过对宫颈癌特异性RARβ基因启动子甲基化水平研究,探讨宫颈癌发生与该基因表达的表观遗传学调控的关系。方法收集维吾尔族妇女宫颈鳞癌(cervical squamous cell carcinoma,CSCC)、宫颈内上皮瘤变(cervical intraepithelial neoplasia,CIN)及宫颈炎患者的新鲜组织标本56例,应用Sequenom MassARRAY甲基化DNA定量分析平台,对组织DNA的RARβ基因启动子区CpG位点进行甲基化水平定量分析。结果 RARβ基因启动子区的目的CpG片段含有16个CpG位点,通过Sequenom MassArray(质谱)对单一CpG位点的甲基化定量测试,发现CpG-6、CpG-12.13(联合位点)和CpG-14等4个CpG位点甲基化率在宫颈癌与宫颈炎组之间有显著差异(P<0.05),并且各个CpG位点的甲基化水平变化存在密切相关性(r>0.5,P<0.01)。但是,从整体CpG片段的甲基化水平角度分析,不同宫颈病变组织之间无统计学差异(P>0.05)。结论 RARβ基因启动子区CpG岛的特定CpG位点高甲基化是宫颈癌演进过程的重要标志,可能与维吾尔族宫颈癌的关系比较密切。从表观遗传学角度分析,此种甲基化引起基因转录水平下调,继而导致蛋白质表达缺失。
Objective To investigate the relationship between cervical carcinogenesis and epigenetic regulation of cervical cancer by studying the promoter methylation of RARβ gene. Methods 56 cases of fresh squamous cell carcinoma of cervical squamous cell carcinoma (CSCC), cervical intraepithelial neoplasia (CIN) and cervicitis were collected from Uyghur women. Sequenom MassARRAY methylation quantitative analysis platform The methylation level of CpG locus in the promoter region of RARβ gene in tissue DNA was quantitatively analyzed. Results The target CpG fragment in the promoter region of RARβ gene contained 16 CpG sites. Quantitative methylation of a single CpG site by Sequenom MassArray revealed that CpG-6, CpG-12.13 (combined site) and CpG -14 and other four CpG sites were significantly different between cervical cancer and cervicitis group (P <0.05), and there was a close correlation between methylation levels of each CpG site (r> 0.5, P <0.01). However, from the perspective of overall CpG methylation level, there was no significant difference between different cervical lesions (P> 0.05). Conclusion The hypermethylation of specific CpG site in CpG island of promoter region of RARβ gene is an important marker of cervical cancer progression and may be closely related to Uighur cervical cancer. From the perspective of epigenetic analysis, this methylation causes a down-regulation of gene transcription, which in turn leads to a loss of protein expression.