DNA methylation in breast and ovarian cancer

Gynecological (including breast) cancer is the most drastic life-threatening and life quality reducing event occurring for women. Although our understanding of how to best treat these malignancies expanding at a dazzling rate, our success in the prevention and early detection of these diseases lags behind. It is questionable whether treatment of manifest cancer can ever become a successful strategy. It is more likely that a greater number of lives can be saved by early detection and prevention. Therefore, it is urgent that researchers identify easily detectable novel markers (e.g. in serum) with high sensitivity and specifity that can accurately predict the risk for developing invasive cancer. The prerequisites for this is to evaluate whether these markers are present in manifest cancers and whether they can predict relapse of disease and are useful as prognostic markers. Tumorspecific DNA changes seem to be very good candidates, because of high stability (compared to RNA) and because of the ability to amplify these changes: DNA-Methylation is a common mechanism for suppressing tumor-suppressor genes during carcinogenesis (recently demonstrated for p15, p16, E-Cadherin, HIC-1, VHL, gelsolin, hMLH1, estrogen receptor, TIMP-3, GSTP-1, DAP-kinase, calcitonin, and RAR-b2, demonstrated by us). Very recently several groups demonstrated that the methylated DNA of some of these genes could be detected to a high degree in serum or plasma of patients, even in patients harboring non-invasive lesions! To date no methylated DNA has been detected in healthy control probands. MethyLight, a real-time PCR based method is a high throughput assay to measure DNA methylation and has the advantage of higher sensitivity compared to MSP (Methylation Specific PCR). The following points should be worked on in the described project: (1) Evaluation whether methylated DNA of tumor suppressor genes works as a marker of malignant breast and ovarian processes and whether these changes are associated with clinicopathological features and prognosis. (2) Could these changes also be found in the blood of the corresponding patient and can these findings predict relapse of disease? (3) Evaluation of the source of methylated DNA in the blood. (4) Is there a correlation between methylation of specific tumor suppressor genes with expression levels of DNA- methylation regulating genes and are expression levels of these genes associated with clinicopathological features and prognosis? The Norris Comprehensive Cancer Center and Peter Jonesgroup are ideal partners to work on this project, because they are one of the worldsleading groups concerning DNA-methylation, they have developed many tools which will be used in this project and have all facilities available, one needs for this. Our department will support all necessary tissue and serum specimens as well as all patient data needed.