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Differential expression of multiple anti‐apoptotic proteins in epidermis of IGF‐1 transgenic mice as revealed by 2‐dimensional gel electrophoresis/mass spectrometry analysis

Overexpression of insulin‐like growth factor‐1 (IGF‐1) has been associated with a number of human tumors, including breast, colon, lung, and prostate cancers. In previous studies, we found that mice overexpressing human IGF‐1 in the basal layer of the epidermis (BK5.IGF‐1 mice) developed skin tumors following treatment with the skin tumor initiator, 7,12‐dimethylbenz[a]anthracene, indicating that IGF‐1 can act as a skin tumor promoter. In the present study, we employed a proteomics approach of two‐dimensional (2‐D) gel electrophoresis and mass spectrometry to profile differentially expressed proteins in skin epidermis between BK5.IGF‐1 transgenic and nontransgenic littermates. Two‐D gels from each of three transgenic and three age/sex matched wild‐type littermates were compared at two different pH ranges. Differentially expressed protein spots were identified by Bio‐Rad's PDQuest image analysis, in‐gel digested, and analyzed on a MALDI‐TOF MS system. A total of 23 proteins were identified as differentially expressed, 17 of them overexpressed in transgenic mice. These proteins included 14‐3‐3 sigma, galectin‐7, an apoptosis‐related protein, three heat shock proteins, four calcium binding proteins, three proteases or protease inhibitors, one actin regulatory capping protein, and translation initiation factor 5A. The differential expression of GRP78, alpha enolase, and galectin‐7 was verified by 1‐D western blot analysis. Two‐D western blot analyses of alpha enolase and galectin‐7 further revealed that alpha enolase had more than one protein spot dependent on charge. The current data suggest that some of the differentially expressed proteins may play a role in the tumor promoting action of IGF‐1 in mouse skin. © 2007 Wiley‐Liss, Inc.