Oxidases as Breast Cancer Oncogens
Final rept. 1 Jun 1999-31 May 2002
NORTHWESTERN UNIV CHICAGO IL
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This proposal examines the novel concept that H2O2 generating oxidase-mediated reactive oxygen species in breast epithelium contribute to the development of breast cancer. Constructs to express xanthine oxidase under the direction of mouse mammary tumor virus MMTV promoter were generated. Transfections performed to generate cell lines stable expressing the enzyme were unsuccessful hence urate oxidase was used as an alternative. The choice for urate oxidase is based on the fact that we have used this enzyme in previous studies and showed that cells expressing this enzyme reveal the characteristic crystals of this enzyme in peroxisomes. Transgenic mice expressing UOX under the transcriptional control of MMTV promoter were generated. We have injected the construct in fertilized ova and identified five founder mice initially which failed to transmit the transgene. Additional microinjections generated 9 founders. Five of these were found to express the transgene. Southern, Northern and Western analyses showed the presence of the transgene in the mammary, and testicular tissues. We will now initiate studies to examine the role of reactive oxygen species in causing cell death, cell proliferation and neoplastic transformation. In parallel, we are also developing a stable cell line in MCFlOA a non tumorogenic cell breast epithelial cells expressing UOX under the control of MMTV promoter. UOX expressing non tumorogenic cells will be injected into nude mice to develop into carcinomas.
- Medicine and Medical Research