神经母细胞瘤中合子阻滞 1（ZAR1）的非启动子 DNA 高甲基化。

Non-promoter DNA hypermethylation of Zygote Arrest 1 (ZAR1) in neuroblastomas.

机构信息

Department of Pediatric Surgery, Nihon University School of Medicine, Tokyo, Japan.

出版信息

J Pediatr Surg. 2013 Apr;48(4):782-8. doi: 10.1016/j.jpedsurg.2012.08.008.

DOI:10.1016/j.jpedsurg.2012.08.008

PMID:23583134

Abstract

BACKGROUND

The comprehensive methylation analysis of tumor-specific differently methylated regions in malignant melanomas and brain tumors has led to the identification of non-promoter hypermethylation of zygote arrest 1 (ZAR1). To search the non-promoter ZAR1 hypermethylation in neuroblastomas, we analyzed the levels of the methylation and transcript expression of ZAR1.

METHODS

The MassARRAY® EpiTYPER (Sequenom Inc., San Diego, CA, USA) system was optimized to determine the quantitative methylation levels of ZAR1 for 12 neuroblastoma cell lines, 23 neuroblastoma samples and four adrenal samples. ZAR1 expression levels were evaluated through a quantitative, real-time reverse transcription-polymerase chain reaction. The quantitative methylation levels of ZAR1 were subjected to correlation studies with the established markers of progressive disease and outcome.

RESULTS

Strikingly, the hypermethylation of ZAR1 regions and ZAR1 expression levels was observed in the neuroblastoma cell lines and neuroblastoma samples, compared to the adrenal samples. Somatic changes in ZAR1 methylation and ZAR1 expression were found in all three neuroblastoma patients. In the ZAR1 regions, poor-outcome tumors that were MYCN-amplified and/or Stage 3 or 4 and/or the age at diagnosis was≥18months, and/or showed an unfavorable histology were frequently hypermethylated.

CONCLUSION

Our results indicate that the hypermethylation of ZAR1 regions is extremely frequent in neuroblastomas and correlates with established markers of progressive disease and outcome.

摘要

背景

对恶性黑色素瘤和脑肿瘤中肿瘤特异性差异甲基化区域的综合甲基化分析，导致了合子阻滞 1（ZAR1）的非启动子超甲基化的鉴定。为了在神经母细胞瘤中搜索非启动子 ZAR1 超甲基化，我们分析了 ZAR1 的甲基化和转录表达水平。

方法

使用 MassARRAY® EpiTYPER（Sequenom Inc.，圣地亚哥，CA，美国）系统对 12 种神经母细胞瘤细胞系、23 种神经母细胞瘤样本和 4 种肾上腺样本的 ZAR1 甲基化定量水平进行了优化。通过定量实时逆转录聚合酶链反应评估 ZAR1 表达水平。ZAR1 的定量甲基化水平与进展性疾病和预后的既定标志物进行了相关性研究。

结果

令人惊讶的是，与肾上腺样本相比，神经母细胞瘤细胞系和神经母细胞瘤样本中观察到 ZAR1 区域的高甲基化和 ZAR1 表达水平。在所有 3 名神经母细胞瘤患者中都发现了 ZAR1 甲基化和 ZAR1 表达的体细胞变化。在 ZAR1 区域中，不良预后的肿瘤通常是 MYCN 扩增和/或 3 或 4 期和/或诊断时年龄≥18 个月和/或具有不良组织学特征，这些肿瘤的 ZAR1 区域常常发生高甲基化。