[维吾尔族女性TAP基因启动子甲基化与HPV感染所致宫颈病变的关系]

[Relationship between TAP gene promoter methylation and cervical lesions with HPV infection in Uyghur women].

作者信息

Aikuer Guzali, Jiao Zhen, Wu Xiao-chuan, Ma Jun-qi, Abudla Abulizi, Hasim Ayshamgul

机构信息

Department of Biology, Xinjiang MedicaI University, Urumqi, China.

出版信息

Zhonghua Bing Li Xue Za Zhi. 2012 Jul;41(7):438-42. doi: 10.3760/cma.j.issn.0529-5807.2012.07.002.

DOI:10.3760/cma.j.issn.0529-5807.2012.07.002

PMID:22932452

Abstract

OBJECTIVE

To study the relationship between TAP (transporter associated with antigen processing) gene promoter regional methylation level and cervical lesions with HPV infection in Uyghur women.

METHODS

A specialized software was used to design specific primers of CpG island fragments of TAP1 and TAP2 gene promoter for PCR amplification, bisulfitemodified SiHa cancer cell DNA for PCR amplification, cloning and sequencing analysis to obtain the relevant information on the gene base sequence methylation of CpG sites. Seventy-eight fresh cervical tissue samples from Uyghur women with cervicitis (number = 15), cervical intraepithelial neoplasia (CIN, number = 30) and cervical squamous cell carcinoma (number = 33) were collected. The methylation level of TAP1 and TAP2 gene promoter regions was detected using MassArray DNA technology. HPV infection status was determined by HPV gene chips. The relationship between CpG-island methylation of gene promoter regions and HPV infection was then analyzed.

RESULTS

Each TAP1 and TAP2 gene corresponding target fragment contained 23 and 8 CpG sites. There were 5 and 8 CpG sites methylation occurred in SiHa cervical cancer cells genomic DNA respectively. The TAP1 methylation level increased steadily with the severity of cervical lesions. The methylation levels in cervical squamous cell carcinoma and CIN (0.048 ± 0.039 and 0.037 ± 0.026, respectively) were higher than that of normal cervical tissue (0.035 ± 0.029, P < 0.05). Although TAP2 gene methylation level also demonstrated similar changes, the difference however was not statistically significant (P > 0.05). HPV gene chip detected 13 HPV genotypes, with HPV16 infection rate being 66.7% (52/78). The methylated proportion of TAP1 positively correlated with HPV16 infection (χ(2) = 6.08, P = 0.039).

CONCLUSION

TAP1 methylation is a remarkable phenomenon occurring in a range of cervical lesions and significantly associated with cervical HPV infection.

摘要

目的

研究维吾尔族女性中抗原加工相关转运体（TAP）基因启动子区域甲基化水平与HPV感染所致宫颈病变的关系。

方法

使用专业软件设计TAP1和TAP2基因启动子CpG岛片段的特异性引物用于PCR扩增，对经亚硫酸氢盐修饰的SiHa癌细胞DNA进行PCR扩增、克隆及测序分析，以获取CpG位点基因碱基序列甲基化的相关信息。收集78例维吾尔族女性的新鲜宫颈组织样本，其中宫颈炎患者15例、宫颈上皮内瘤变（CIN）患者30例、宫颈鳞状细胞癌患者33例。采用MassArray DNA技术检测TAP1和TAP2基因启动子区域的甲基化水平。通过HPV基因芯片确定HPV感染状态。随后分析基因启动子区域CpG岛甲基化与HPV感染的关系。

结果

每个TAP1和TAP2基因对应的目标片段分别包含23个和8个CpG位点。在SiHa宫颈癌细胞基因组DNA中分别有5个和8个CpG位点发生甲基化。TAP1甲基化水平随宫颈病变严重程度而稳步升高。宫颈鳞状细胞癌和CIN中的甲基化水平（分别为0.048±0.039和0.037±0.026）高于正常宫颈组织（0.035±0.029，P<0.05）。虽然TAP2基因甲基化水平也呈现类似变化，但差异无统计学意义（P>0.05）。HPV基因芯片检测到13种HPV基因型，HPV16感染率为66.7%（52/78）。TAP1的甲基化比例与HPV16感染呈正相关（χ(2)=6.08，P=0.039）。