High throughput detection of M6P/IGF2R intronic hypermethylation and LOH in ovarian cancer

Nucleic Acids Res. 2006 Jan 23;34(2):555-63. doi: 10.1093/nar/gkj468. Print 2006.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Alleles
  • CpG Islands*
  • DNA Methylation*
  • Female
  • Humans
  • Introns
  • Loss of Heterozygosity*
  • Ovarian Neoplasms / genetics*
  • Polymerase Chain Reaction / methods*
  • Receptor, IGF Type 2 / genetics*
  • Sequence Analysis, DNA
  • Sulfites / chemistry


  • Receptor, IGF Type 2
  • Sulfites
  • sodium bisulfite