Proteomic analysis of the small intestine and colon epithelia of adenomatous polyposis coli gene-mutant mice by two-dimensional gel electrophoresis.
Mutations of the adenomatous polyposis coli gene (APC) have been implicated in the occurrence of sporadic colon cancer. Various APC mutant strains of mice have been created to better understand the function of this gene. Previously, we had mice express a mutant form of mRNA of the APC protein that encoded 474 amino acids instead of the 2845 amino acids due to exon duplication. These APC mutant mice (APC delta 474) developed intestinal and mammary tumors, as have other APC mutant mice previously reported (Sasai, H., et al. Carcinogenesis, in press). To elucidate the mechanism of the tumor development, we prepared protein samples from both normal and tumor tissues from APC delta 474 mutant mice, as well as tissues from normal mice, and used them for proteomic analysis. After two-dimensional electrophoresis, the gels were silver stained and the protein spots were analyzed. We analyzed about 1000 protein spots per sample and found several protein spots that are specific for normal or tumor samples from APC delta 474 mutant mice, as well as proteins with altered expression levels. Among the identified protein spots, truncated beta-tubulins were specific to APC delta 474 mutant mice polyp samples. The apparent molecular mass of these proteins suggested that these beta-tubulins may be truncated very close to the binding site of the anti-tumor drug taxol.
Minowa
T
T
Japan Tobacco Inc., Pharmaceutical Frontier Research Laboratories, Central Pharmaceutical Research Institute, Yokohama, Kanagawa. tminowa@asahi-net.email.ne.jp
Ohtsuka
S
S
Sasai
H
H
Kamada
M
M
D011125:Adenomatous Polyposis Coli / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D003106:Colon / Q000737:chemistry* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D017491:Genes, APC; D007413:Intestinal Mucosa / Q000737:chemistry* / Q000473:pathology; D007421:Intestine, Small / Q000737:chemistry* / Q000473:pathology; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*
D009363:Neoplasm Proteins; D020543:Proteome