摘要
目的观察乳腺癌中DNA结合抑制因子4(ID4)基因启动子区甲基化状态及其与临床病理特征间的关系,探讨ID4基因在乳腺癌发病过程中的作用机制。方法采用焦磷酸测序法定量检测乳腺癌及正常组织标本中ID4基因启动子区甲基化水平,并分析其在不同临床病理特征间的差异;用RT-PCR检测MM-453细胞去甲基化处理前后ID4启动子区甲基化水平及mRNA表达情况。结果乳腺癌组织中ID4启动子区甲基化水平显著高于正常乳腺组织[(31.16±1.50)%vs(19.89±0.22)%,P<0.01];ER+乳腺癌组织中ID4甲基化水平显著高于ER-乳腺癌组织[(36.57±1.97)%vs(27.91±1.83)%,P<0.01];去甲基化处理后MM-453细胞ID4甲基化水平明显下降,mRNA表达显著上升,两者呈负相关(r=-0.973,P<0.01)。结论 ID4基因可能通过启动子区高甲基化等多种途径在乳腺癌发生、发展过程中起着重要作用,其启动子区高甲基化在ER+的乳腺癌中具有重要意义。
Objective To explore the possible mechanism of inhibitor of DNA binding 4 （ID4） gene in breast tumorigenesis by observ- ing the correlations of methylation status of//）4 gene promoter in breast cancer cells with different clinical pathological characteristics. Methods The methylation levels of//）4 promoter region in breast tumor tissue （n = 40） and normal tissue specimens （n = 20） were determined by pyrosequencing and the correlations between the methylation level of ID4 gene and clinical pathological characteristics were analyzed. The methylation levels of ID4 promoter region in MM-453 cell line before and after demethylation treatment were deter- mined by pyrosequencing and the mRNA expression of ID4 was determined by RT-PCR. Results The methylation level of ID4 promoter region in breast tumor tissue was significantly higher than that in normal breast tissue [ （31.16 ±1.50） % vs （ 19.89 ± 0.22） %, P 〈0.01 ]. The methylation level of ID4 gene in ER-positive breast cancer tissue showed significantly higher than that in ER-negative group[ （36.57 ± 1.97） % vs （27.91 ± 1.83） % ,P 〈0.01 ]. The re-expression of 1/）4 gene mRNA was significantly increased and negatively correlated with the methylation of 11）4 in MM-453 cell line （ r = - 0.973, P 〈 0.01 ）. Conclusion ID4 gene may play an important role in breast tumorigenesis and development by a variety of way including hypermethylation of gene promoter, especially in ER-positive breast cancer.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2014年第8期580-583,共4页
Chinese Journal of Clinical Laboratory Science
基金
江苏省卫生厅医学领军人才暨创新团队项目(LJ201131)
江苏省科技厅临床医学专项(BL2013036)
