Biallelic methylation and silencing of paternally expressed gene 3 (PEG3) in gynecologic cancer cell lines

Objective. To measure mRNA expression levels of paternally expressed gene 3 (PEG3) in gynecologic cancer cell lines and to determine if DNA methylation is involved in the control of PEG3 expression. Methods. PEG3 mRNA levels were measured with real-time PCR from 28 gynecologic cancer cell lines and compared to normal tissues. PEG3 mRNA expression was correlated to promoter methylation levels measured by real-time methylation-specific PCR. Polymorphism-specific restriction digestion was employed to analyze PEG3 allele distribution. Results. While expressed in normal gynecologic tissues, PEG3 is silenced in all endometrial and cervical cancer cell lines studied. In the eight ovarian cancer cell lines, five were found to be PEG3 negative, the remaining three express low levels of PEG3 mRNA. In contrast, loss of maternal imprinting and relatively high PEG3 expression levels were detected in all four choriocarcinomas cell lines studied. No cell line confirmed to contain two copies of PEG3 expressed PEG3 mRNA, suggesting that PEG3 downregulation is not due to genetic deletion. PEG3 mRNA expression was, however, quantitatively correlated to its promoter methylation status. Treatment of PEG3 negative cells with DNA methyltransferase inhibitor 5′-aza-deoxycytidine led to partial promoter demethylation and biallelic reactivation of PEG3 transcription, confirming the methylation-mediated mechanism for PEG3 silencing. Conclusion. PEG3 silencing is associated with DNA hypermethylation but not gene deletion in cell lines tested. These results suggest that loss of PEG3 expression may be a frequent event in gynecologic cancers. Given the known role of PEG3 in p53-mediated apoptosis, it is possible that PEG3 functions as a tumor suppressor.