Microarray and real time-polymerase chain reaction analysis of mammary glands from rats fed conjugated linoleic acid
Smonskey, Matthew Thomas
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Conjugated linoleic acid (CLA) prevents mammary tumors in rats. The purpose of this study was to test the hypothesis that common pathways are altered by two similarly chemoprotective isomers, 9,11-CLA and 10,12-CLA. Microarray analysis of rat mammary glands after CLA feeding demonstrated altered expression of genes involved cellular growth, death, and cancer development. Validation by real-time PCR was performed on mammary tissues for the following genes: Adam17, Agt, Bin1, Btg2, Casp2, Dgka, Ets1, Gas5, Hmgb2, Krt4, Psmb9, Stat1, Tp53, and Vdac1. Most genes validated by RT-PCR showed a pattern of change similar to that seen by microarray. Lastly, a study was conducted to determine if whole gland changes are represented in a fibroblast population. RT-PCR of CLA-treated rat mammary stromal vascular cells demonstrated a distinct pattern of gene expression from whole gland, suggesting that fibroblasts may not be the primary target of CLA induced changes observed in the microarray.