Lung cancer is responsible for the most cancer-related mortality worldwide and the mechanism of its development is poorly understood. Proteomics has become a powerful tool offering vital knowledge related to cancer development. Using a two-dimensional difference gel electrophoresis (2D-DIGE) approach, we sought to compare tissue samples from non-small-cell lung cancer (NSCLC) patients, exactly adenocarcinoma (ADC), taken from the tumor center and tumor margin and control (adjacent non-tumor) tissues . We found 22 differentially abundant spots representing 17 proteins differentiating center and margin. Sixty eight proteins; represented by 79 spots :40, 20, and 19 spots differed between the control and center and margin, control and center, and control and margin, respectively. Twenty six significant canonical pathways were identified, including Rho signaling pathways, a semaphorin neuronal repulsive signaling pathway, and epithelial adherens junction signaling. Proteins differentiating the tumor center and tumor margin were linked to cancer invasion and progression, including cell migration, adhesion and invasion, cytoskeletal structure, protein folding, anaerobic metabolism, tumor angiogenesis, EMC transition, epithelial adherens junctions, and inflammatory responses.