Division of Cutaneous Science, Department of Dermatology, Nihon University Graduate School of Medicine, Tokyo 173-8610, Japan.
J Dermatol Sci. 2010 Aug;59(2):98-106. doi: 10.1016/j.jdermsci.2010.04.016. Epub 2010 May 20.
Epigenetic changes such as aberrant DNA methylation and histone modification have been shown to play an important role in the tumorigenesis of malignant melanoma.
To identify novel tumor-specific differentially methylated regions (DMRs) in human malignant melanoma.
The aberrant methylation at 14 candidate human genomic regions identified through a mouse model study with quantitative DNA methylation analysis using the Sequenom MassARRAY system was performed.
The CpG island Exon 1 region of the Zygote arrest 1 (ZAR1) gene, which is responsible for oocyte-to-embryo transition, showed frequent aberrant methylation of 28 out of 30 (93%) melanoma surgical specimens, 16 of 17 (94%) melanoma cell lines, 0% of 4 normal human epidermal melanocyte (NHEM) cell lines, 0% of 10 melanocytic nevi and 100% of 51 various cancer cell lines. According to the real-time RT-PCR, the ZAR1 gene was overexpressed in part of the hypermethylated cell lines, while its low expression with bivalent histone methylation status was seen in unmethylated cell lines.
Our findings suggest that the ZAR1 intra-genic differentially methylated region would be a useful tumor marker for malignant melanoma and may be other type of cancers. The involvement of ZAR1 in the carcinogenesis of melanoma, still remains unclear, although we have examined tumorigenic capacities by exogenous full-length ZAR1 over-expression and siRNA knock-down experiments.
表观遗传改变,如异常的 DNA 甲基化和组蛋白修饰,已被证明在恶性黑色素瘤的肿瘤发生中起重要作用。
鉴定人类恶性黑色素瘤中新型肿瘤特异性差异甲基化区域(DMR)。
通过定量 DNA 甲基化分析,使用 Sequenom MassARRAY 系统在小鼠模型研究中鉴定出 14 个候选人类基因组区域的异常甲基化,对这些区域进行了分析。
卵母细胞向胚胎过渡所必需的 Zygote arrest 1(ZAR1)基因的 CpG 岛外显子 1 区域,在 30 个(93%)黑色素瘤手术标本、17 个(94%)黑色素瘤细胞系、4 个(0%)正常人类表皮黑素细胞(NHEM)细胞系、10 个(0%)黑素细胞痣和 51 个(100%)各种癌细胞系中,有 28 个出现了频繁的异常甲基化。根据实时 RT-PCR,部分高甲基化细胞系中 ZAR1 基因过表达,而在未甲基化细胞系中则表现出二价组蛋白甲基化状态的低表达。
我们的研究结果表明,ZAR1 基因内差异甲基化区域可能是恶性黑色素瘤和其他类型癌症的有用肿瘤标志物。虽然我们已经通过外源性全长 ZAR1 过表达和 siRNA 敲低实验研究了 ZAR1 在黑色素瘤发生中的致癌作用,但仍不清楚其具体作用机制。