DNA-methylation changes in premalignant lesions of the breast. This project is funded by the Pennsylvania Breast Cancer Coalition and Friends for an Earlier Breast Cancer Test

Reversion of Phenotype

In this project, we are aiming to determinate the methylation pattern of premalignant lesions like ductal hyperplasia (DH), atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS).  We want to determine if the methylation pattern detected in the breast tissue samples can be detected in the DNA recovered from the serum of women carrying these lesions. The use of body fluids such as serum offers a novel approach to the early detection by a non-invasive sampling in patients at risk of cancer. We are interested to define the DNA methylation profile of the different lesions and define a panel of biomarkers of breast cancer risk.  

For more information about the Pennsylvania Breast Cancer Coalition, visit pabreastcancer.org. And for more information about Friends for an Earlier Breast Cancer Test, visit earlier.org.

Reversion of the neoplastic transformation

Results

This project aims to correlate the data on clinical samples with “in vitro” studies for finding “solutions” that could be translated to cancer prevention and therapy in the clinical setting. For this purpose we are using an “in vitro” model of breast cancer progression developed in the BCRL for such analysis. The MCF-10F progression model consists of four derived cell lines: (a) the spontaneously immortalized cell line MCF-10F, which does not show any characteristic of invasiveness or tumor formation and therefore is considered to be a normal-like breast epithelial cell line; (b) the transformed trMCF cells; (c) the invasive bsMCF cell line and; (d) the tumor cell lines, caMCFs, which shown all characteristics of a fully malignant breast cancer cell types. In 3D-cultures, normal cells such as MCF-10F form tubules although transformed cells such as trMCF form solid masses.  The bsMCF cells induced tumors in SCID mice that were poorly differentiated adenocarcinomas that were ESRα, progesterone receptor (PR) and ERBB2 negatives. Using this model, DNA-methylation studies were performed and we have found genes, such as NRG1 and RARβ, that in the normal cells were unmethylated became hypermethylated at the invasive and tumorigenic stages. Retinoids are the ligands of RARβ and they have been recognized as promising anticancer compounds. Using this in vitro model, I want to study the potential of retinoic acid to revert the neoplastic process. We found that after treatment of trMCF with retinoic acid, there was a reduction of solid masses and increased of tubules in collagen indicating that retinoic acid was able to revert the transformed phenotype of trMCF. The trMCF cells would represent a very early stage in the transformation resembling premalignant lesions. In the future, I’m planning to perform studies using animal models to have a clear picture of the capacities of retinoic acid in this process.