Abbreviations used: ATP -adenosine triphosphate; DTT -dithiothreitol; ECMextracellular matrix; GDT -guanosine diphosphate; LR -log ratio; MALDI TOF-MS/MS -Matrix-assisted laser-desorption ionization time-of-flight tandem mass spectroscopy; MMPs -matrix metalloproteinases; PAGE -polyacrylamide gel electrophoresis; PBSphosphate buffered solution; PCR -polymerase chain reaction; SDS -sodium dodecyl sulfate; TCA -trichloroacetic acid; TCTP -translationally controlled tumor protein; 2-DE -two-dimensional gel electrophoresis Abstract: Tumor establishment and penetration consists of a series of complex processes involving multiple changes in gene expression and protein modification. Proteome changes of tumor tissue were investigated after intraperitoneal administration of a high concentration of ascorbic acid in BALB/C mice implanted with CT-26 cancer cells using two-dimensional gel electrophoresis and mass spectrometry. Eighteen protein spots were identified whose expression was different between control and ascorbic acid treatment groups. In particular, eukaryotic translation initiation factor 3 subunit 1, nucleophosmin, latexin, actin-related protein 2/3 complex subunit 5, M2-type pyruvate kinase, vimentin, tumor protein translationally-controlled 1, RAS oncogene family Ran, plastin 3 precursor, ATPase, Rho GDT dissociation inhibitor β, and proteasome activator subunit 2 expression were quantitatively up-regulated. The increase in the level of these proteins was accompanied by an increase in mRNA level. The cytoskeleton protein actin, vimentin, and tumor protein translationally-controlled 1 showed quantitative expression profile differences. A change in actin cytoskeleton distribution, functionally relevant to the proteome result, was observed after treatment with ascorbic acid. These