High-throughput detection of aberrant imprint methylation in the ovarian cancer by the bisulphite PCR-Luminex method

In the current study, we applied the BPL method to the analysis of DNA methylation for identification of prognostic panels of DNA methylation cancer biomarkers of imprinted genes. We found that the BPL method precisely quantified the methylation status of specific DNA regions in somatic cells. We found a higher frequency of LOI than LOH. LOI at IGF2, PEG1 and H19 were frequent alterations, with a tendency to show a more hypermethylated state. We detected changes in DNA methylation as an early event in ovarian cancer. The degree of LOI (LOH) was associated with altered DNA methylation at IGF2/H19 and PEG1.The relative ease of BPL method provides a practical method for use within a clinical setting. We suggest that DNA methylation of H19 and PEG1 differentially methylated regions (DMRs) may provide novel biomarkers useful for screening, diagnosis and, potentially, for improving the clinical management of women with human ovarian cancer.Human ovarian cancer (HOC) is the leading cause of death from gynecological malignancies, primarily due to the lateness of detection when the cancer is already at an advanced stage. Effective screening protocols for early stages are not currently available. HOC is characterized by complex genetic and epigenetic alterations, including loss of heterozygosity (LOH) and loss of imprinting (LOI) [1,2]. Such alterations are presumed to represent the second hit, according to Knudson's two-hit hypothesis (OMIM #167000) [3]. However, alterations in DNA methylation can also occur as the first hit during human carcinogenesis [4].For childhood cancers such as retinoblastoma (OMIM #180200), Wilms' tumor (OMIM #194070) and osteosarcoma (OMIM #259500), changes primarily occur on the paternal allele first, followed by a second hit on the maternal allele [5,6]. Complete hydatidiform moles, which are of androgenetic or paternal origin, are characterized by malignant transformation whereas ovarian teratomas, which are of parthenogenetic or maternal origin