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The Role of Oxidative Stress in Apoptosis of Breast Cancer.

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Annual rept. 29 Aug 94-28 Aug 95,

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Apoptosis is a biological process for cell removal, for which the underlying mechanism is not well understood. Restoring the pathways that control apoptosis holds promise as a new approach for the treatment of breast cancer. The objective of the research project is to test the hypothesis that oxidative stress plays a critical role in the mechanism of apoptosis induced by treatment of human breast cancer cell lines with tumor necrosis factor-a ThF-a. The goal of the first year of the project was to use Northern blot hybridization analyses and enzyme assays to defme the extent of changes in the cellular antioxidant defense in ThF-a-treated breast cancer cells. Our first studies demonstrated dose- and time-dependent increases in apoptosis, based on an ELISA for DNA fragments, with TNF-a treatment of MCF-7 cells. In conducting Northern blot hybridization analysis, we encountered problems that appeared to be due to UV damage to the cDNA probe templates, that occmred while puritying them. The problem was resolved by shielding the sample during the workup. For biochemical measurements of the antioxidant defense, we have a catalase assay underway and are comparing methods for measuring OSH levels.

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  • Biochemistry
  • Medicine and Medical Research

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