The effect of mitochondrial uncoupling in murine skin samples using Keratin 5-UCP3 trangenic model. Mus musculus

Uncoupling protein 3 (UCP3) is a nuclear encoded, member of the mitochondrial solute carrier family which localizes to the inner mitochondrial membrane and dissipates the proton gradient used to drive ATP synthesis. UCP3 is expressed at low levels in skin and our lab has demonstrated that overexpression of UCP3 in epidermis (K5-UCP3 mice) provides marked resistance to two-stage chemical carcinogenesis protocols in skin. To further understand the mechanisms by which UCP3 promotes this cancer resistant phenotype, we performed a microarray analysis of dorsal skin biopsies from wild type FVB/N and K5-UCP3 mice. The results of this array indicate that UCP3 overexpression in skin enhances keratinocytes differentiation and fatty acid oxidation in skin, and taken together with other data we have provided a mechanistic link between fatty acid oxidation and Akt inhibition in UCP3-mediated cancer resistance. Overall design: Whole skin biopsies were taken from dorsal skin of 7 week old male wild type FVB/N (n=3) and K5-UCP3 mice (n=3) and RNA was isolated using Trizol Reagent followed by clean up using the Qiagen Rneasy Kit. Samples were amplified from 100ng RNA using the Illumina RNA Amplification Kit. Biotin-16-UTP was used to label transcripts which were then hybridized (700ng per array) to the Illumina Mouse-6 v1.0 BeadChip. Arrays were scanned with an Illumina bead array reader confocal scanner. Array data processing was performed using the Illumina BeadStudio software.