High throughput detection of M6P/IGF2R intronic hypermethylation and LOH in ovarian cancer
High throughput detection of M6P/IGF2R intronic hypermethylation and LOH in ovarian cancer
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DOI：
10.1093/nar/gkj468
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发表时间：
2006-01-01
影响因子：
14.9
通讯作者：
Murphy, SK
中科院分区：
文献类型：
--
作者：
Huang, ZQ;Wen, YQ;Murphy, SK
细胞表面甘露糖6-磷酸/胰岛素样生长因子II受体（M6P/IGF2R）结合并靶向外源性胰岛素样生长因子II （IGF2）到前溶酶体，在那里它被降解。M6P/IGF2R的杂合性缺失（LOH）在癌症中发现，剩余等位基因突变失活。我们利用M6P/IGF2R内含子2 CpG岛的正常等位基因特异性差异甲基化来快速评估卵巢癌中潜在的LOH，因为每个正常个体都有信息。为此，我们开发了一种亚硫酸盐修饰基因组DNA的96孔格式方法，允许快速甲基化分析。我们发现了M6P/IGF2R LOH的卵巢癌，但出乎意料的是，我们还发现在内含子2的父系遗传等位基因上经常出现甲基化异常。这些结果证明了我们的亚硫酸酯改性高通量方法对大样本数分析的实用性。他们进一步表明，内含子2 CpG岛的甲基化状态可能是LOH的有用指标和疾病的生物标志物。
Cell surface mannose 6-phosphate/insulin-like growth factor II receptors (M6P/IGF2R) bind and target exogenous insulin-like growth factor II (IGF2) to the prelysosomes where it is degraded. Loss of heterozygosity (LOH) for M6P/IGF2R is found in cancers, with mutational inactivation of the remaining allele. We exploited the normal allele-specific differential methylation of the M6P/IGF2R intron 2 CpG island to rapidly evaluate potential LOH in ovarian cancers, since every normal individual is informative. To this end, we developed a method for bisulfite modification of genomic DNA in 96-well format that allows for rapid methylation profiling. We identified ovarian cancers with M6P/IGF2R LOH, but unexpectedly also found frequent abnormal acquisition of methylation on the paternally inherited allele at intron 2. These results demonstrate the utility of our high-throughput method of bisulfite modification for analysis of large sample numbers. They further show that the methylation status of the intron 2 CpG island may be a useful indicator of LOH and biomarker of disease.