Comprehensive analysis of MGMT promoter methylation: correlation with MGMT expression and clinical response in GBM.
Comprehensive analysis of MGMT promoter methylation: correlation with MGMT expression and clinical response in GBM.
复制标题
DOI：
10.1371/journal.pone.0016146
复制
发表时间：
2011-01-07
期刊：
影响因子：
3.7
通讯作者：
Foltz G
中科院分区：
文献类型：
--
作者：
Shah N;Lin B;Sibenaller Z;Ryken T;Lee H;Yoon JG;Rostad S;Foltz G
O 6-甲基鸟嘌呤DNA-甲基转移酶（MGMT）启动子甲基化已被确定为胶质母细胞瘤患者的潜在预后标志物。启动子甲基化的确切位点与其对基因沉默的影响以及患者随后对治疗的反应之间的关系仍在确定中。本研究的目的是全面表征整个MGMT启动子的胞嘧啶-鸟嘌呤（CpG）二核苷酸甲基化，并将单个CpG位点甲基化模式与mRNA表达、蛋白质表达和无进展生存期相关联。为了最好地确定最能预测基因沉默和对治疗反应的特定MGMT启动子区域，我们使用定量亚硫酸氢盐测序法测定了70例GBM患者肿瘤样本中MGMT启动子中所有97个CpG位点的甲基化状态。我们接下来鉴定了最能预测基因沉默和改善无进展生存的CpG位点特异性和区域甲基化模式。使用这些数据，我们提出了一种新的分类方案，利用整个启动子的甲基化数据，并表明基于这种方法的分析，我们称之为3R分类，可以预测无进展生存期（HR = 5.23，95%CI [2.089-13.097]，p<0.0001）。 为了使该方法适应临床环境，我们使用了基于3R分类的甲基化特异性多重连接依赖性探针扩增（MS-MLPA）检测，并表明该检测在临床环境中可行，可预测无进展生存期（HR = 3.076，95% CI [1.301-7.27]，p = 0.007）。 我们讨论了潜在的优势，测试的基础上，这种启动子范围的分析，并将其与常用的甲基化特异性PCR检测。这两种方法在一个大的独立患者队列中的进一步前瞻性验证将需要确认在临床环境中MGMT甲基化的启动子范围分析的附加值。
O6-methylguanine DNA-methyltransferase (MGMT) promoter methylation has been identified as a potential prognostic marker for glioblastoma patients. The relationship between the exact site of promoter methylation and its effect on gene silencing, and the patient's subsequent response to therapy, is still being defined. The aim of this study was to comprehensively characterize cytosine-guanine (CpG) dinucleotide methylation across the entire MGMT promoter and to correlate individual CpG site methylation patterns to mRNA expression, protein expression, and progression-free survival. To best identify the specific MGMT promoter region most predictive of gene silencing and response to therapy, we determined the methylation status of all 97 CpG sites in the MGMT promoter in tumor samples from 70 GBM patients using quantitative bisulfite sequencing. We next identified the CpG site specific and regional methylation patterns most predictive of gene silencing and improved progression-free survival. Using this data, we propose a new classification scheme utilizing methylation data from across the entire promoter and show that an analysis based on this approach, which we call 3R classification, is predictive of progression-free survival (HR = 5.23, 95% CI [2.089–13.097], p<0.0001). To adapt this approach to the clinical setting, we used a methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) test based on the 3R classification and show that this test is both feasible in the clinical setting and predictive of progression free survival (HR = 3.076, 95% CI [1.301–7.27], p = 0.007). We discuss the potential advantages of a test based on this promoter-wide analysis and compare it to the commonly used methylation-specific PCR test. Further prospective validation of these two methods in a large independent patient cohort will be needed to confirm the added value of promoter wide analysis of MGMT methylation in the clinical setting.
登录
查看更多内容
影响因子：
3.9
作者：
Kitange, Gaspar J.;Carlson, Brett L.;Mladek, Ann C.;Decker, Paul A.;Schroeder, Mark A.;Wu, Wenting;Grogan, Patrick T.;Giannini, Caterina;Ballman, Karla V.;Buckner, Jan C.;James, C. David;Sarkaria, Jann N.
通讯作者：
Sarkaria, Jann N.
影响因子：
4.8
作者：
Cao, Van Thang;Lung, Tae-Young;Chae, Hong-Jae
通讯作者：
Chae, Hong-Jae
影响因子：
6.4
作者：
Grasbon-Frodl, Eva M.;Kreth, Friedrich Wilhelm;Kretzschmar, Hans A.
通讯作者：
Kretzschmar, Hans A.
影响因子：
4.1
作者：
Mikeska, Thomas;Bock, Christoph;Waha, Andreas
通讯作者：
Waha, Andreas
影响因子：
158.5
作者：
Hegi, ME;Diserens, A;Stupp, R
通讯作者：
Stupp, R