Breast Cancer Prevention by Inducing Apoptosis in DCIS Using Breast Ductal Lavage

Brief Summary

Detailed Description

OBJECTIVES: * Determine the expression pattern of the programmed cell death (PCD) regulatory genes bcl-2, bax, and bcl-xL in primary ductal carcinoma in situ (DCIS) cultures. * Determine whether down-regulation by genetic manipulation of the anti-apoptotic genes bcl-2 and/or bcl-xL, alone or in conjunction with physiological preventive doses of tamoxifen citrate, has the highest induction of PCD in primary DCIS cell cultures. * Determine the expression pattern of the PCD regulatory genes bcl-2, bax, and bcl-xL in cells obtained by breast ductal lavage. * Determine whether down-regulation by genetic manipulation of the anti-apoptotic genes bcl-2 and/or bcl-xL, alone or in conjunction with physiological preventive doses of tamoxifen citrate, has the highest induction of PCD in cells obtained by breast ductal lavage. OUTLINE: Patients undergo breast lavage to collect primary epithelial cells for cytological analysis before a planned surgical procedure. Ductal carcinoma in situ (DCIS) tissue samples obtained from surgery are used to establish primary DCIS cell cultures. The DCIS cells and primary epithelial cells obtained by ductal lavage are analyzed for endogenous protein levels of bcl-2, bax, and bcl-xL, using western blotting and immunohistochemical staining, to determine the appropriate antisense oligonucleotide molecule that will be used to induce apoptosis. The DCIS cells and primary epithelial cells obtained by ductal lavage are treated with antisense oligonucleotides and/or a physiological chemopreventive dose of tamoxifen citrate to determine which will provide the highest induction of cell death. The effect of these treatments on protein expression is analyzed by western blotting and immunohistochemistry. The effect of these treatments on markers of programed cell death (PCD) (i.e., DNA fragmentation and caspase activation) is also analyzed. Changes in mRNA expression are analyzed using a PCR-based quantitation assay. Results from the molecular marker assays are not provided to the patients.

Primary Outcomes