title: High throughput detection of M6P/IGF2R intronic hypermethylation and LOH in ovarian cancer.
abstract: 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.
authors.affiliation: Division of Gynecologic Oncology, Duke University Medical Center, Durham, NC 27708, USA.
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Huang
authors.forename: Yaqing
authors.initials: Y
authors.lastname: Wen
authors.forename: Ruby
authors.initials: R
authors.lastname: Shandilya
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Marks
authors.forename: Andrew
authors.initials: A
authors.lastname: Berchuck
authors.forename: Susan K
authors.initials: SK
authors.lastname: Murphy
mesh_terms: D000483:Alleles; D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D006801:Humans; D007438:Introns; D019656:Loss of Heterozygosity; D010051:Ovarian Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017527:Receptor, IGF Type 2 / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013447:Sulfites / Q000737:chemistry
chemical_list: D017527:Receptor, IGF Type 2; D013447:Sulfites; C009279:sodium bisulfite