Promoter Switching and Transcription Factor Usage During Breast Adipocyte Differentiation Role in Aromatase Expression and Activity

摘要

During the last 12-months, most work proposed under Specific Aim 1 was substantially completed. Aim 1: aromatase mRNA translational efficiency will be determined by, monitoring its distribution in both translationally active polysomes and translationally inactive monosome or messenger ribonucleotide particles (mRNPs). This approach will enable us to establish if promoter switching allows a given aromatase mRNA species to be translated more efficiently, thereby resulting in higher amounts of aromatase protein under these circumstances. Cellular amounts of aromatase mRNA-total and Promoter- specific will be determined by competitive reverse transcriptase polymerase chain reaction (RT-PCR). A murine cell-culture model of fibroblast-to-adipocyte differentiation was validated and used to demonstrate aromatase gene promoter- switching. Validation of a human cell culture model to be used in further studies is nearly complete. The pace of progress was adversely affected by the loss of a technician who departed in February 2004 to relocate out of state. Efforts to find a suitably qualified replacement were unsuccessful until October 2004. Completion of work proposed under Specific Aim 1, and Aims 2 and 3 during the current no-cost extension period will be greatly facilitated by the addition of a technician who has already been engaged to start work on November 29, 2004.