Identification of PRTFDC1 silencing and aberrant promoter methylation of GPR150, ITGA8 and HOXD11 in ovarian cancers
Identification of PRTFDC1 silencing and aberrant promoter methylation of GPR150, ITGA8 and HOXD11 in ovarian cancers
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DOI：
10.1016/j.lfs.2007.01.015
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发表时间：
2007-03-27
期刊：
影响因子：
6.1
通讯作者：
Ushijima, Toshikazu
中科院分区：
文献类型：
--
作者：
Cai, Li-yi;Abe, Masanobu;Ushijima, Toshikazu
甲基化启动子CpG岛（CGIs）可用于发现新的肿瘤抑制基因和疾病标志物。在这项研究中，以确定启动子CGIs异常甲基化在人类卵巢癌，我们进行了全基因组筛选差异甲基化的DNA片段甲基化敏感的代表性差异分析（MS-RDA）。MS-RDA从卵巢癌细胞系（ES-2）和正常人卵巢上皮细胞系（HOSE 6 -3）中分离出185个特异性甲基化的DNA片段，其中33个来自推定的启动子CGIs。通过甲基化特异性PCR分析了10个卵巢癌细胞系，并且33个CGI中的7个（GPR 150、L0 C222171、PRTFDC 1、L0 C339210、ITGA 8、C9 orf 64和HOX 1911）在一个或多个细胞系中被甲基化。通过定量逆转录-PCR，它们的下游基因在没有未甲基化DNA分子的细胞系中几乎不表达。用5-氮杂-2 '-脱氧胞苷对甲基化细胞系进行去甲基化，恢复了两种基因（PRTFDC 1和C9 orf 64）的表达。在原发性卵巢癌中，GPR 150（15种癌症中的4种）、ITGA 8（2/15）、PRTFDC 1（1/15）和HOAD II（1/15）的CGI被甲基化。本研究首次发现PRTFDC 1基因沉默，GPR 150、ITGA 8和HOXD 11基因异常甲基化可能是候选的肿瘤标志物。(c)2007爱思唯尔公司All rights reserved.
Methylated promoter CpG islands (CGIs) can be used to find novel tumor-suppressor genes and disease markers. In this study, to identify promoter CGIs aberrantly methylated in human ovarian cancers, we performed a genome-wide screening for differentially methylated DNA fragments using methylation-sensitive-representational difference analysis (MS-RDA). MS-RDA isolated 185 DNA fragments specifically methylated in an ovarian cancer cell line (ES-2), compared with a normal human ovarian surface epithelial cell line (HOSE6-3), and 33 of them were derived from putative promoter CGIs. Ten ovarian cancer cell lines were analyzed by methylation-specific PCR, and seven (GPR150, LOC222171, PRTFDC1, LOC339210, ITGA8, C9orf64 and HOX1911) of the 33 CGIs were methylated in one or more of the cell lines. Their downstream genes were barely expressed in cell lines without unmethylated DNA molecules by quantitative reverse-transcription-PCR. Demethylation of methylated cell lines with 5-aza-2'-deoxycytidine restored expression of two genes (PRTFDC1 and C9orf64). In primary ovarian cancers, CGIs of GPR150 (in 4 of 15 cancers), ITGA8 (2/15), PRTFDC1 (1/15), and HOADII (1/15) were methylated. Silencing of PRTFDC1 was revealed here for the first time, and aberrant methylation of GPR150, ITGA8 and HOXD11 could be candidate tumor markers. (c) 2007 Elsevier Inc. All rights reserved.