MASPIN, which is also known as Serpin B5, is a novel tumor suppressor. Emerging evidence suggests that MASPIN acts as a multifaceted protein in various types of cancer, including prostate, breast and pancreatic cancer. It interacts with diverse groups of intercellular and extracellular proteins, regulating cell adhesion, motility, apoptosis and angiogenesis, and is involved in mammary gland development. As MASPIN is a multifunctional factor in cancer pathways, its function remains poorly illuminated. In this study, we compared the protein profiles of LC5 cell lines with MASPIN overexpression and knockdown using comparative two-dimensional gel electrophoresis. The differences in protein expression, visualized as differences in spots, were identified by time-of-flight (TOF)/TOF mass spectometry. Significant differences were observed between overexpressing and knocked down cells, including eight spots that were unique and sixteen spots that were up- or down-regulated by more than 4-fold. Six genes, including Sdccag8, Ldoc1, SCAI, SDCCAG3, CT62 and NEDD9 were unique in MASPIN-expressing cell lines, but absent in knock-out cell lines, in which most of them play a significant role in the invasion of cancer cells. Moreover, the Brms1 and CAGE1 genes were identified as being uniquely expressed in knocked down cell lines, which were associated with the development and progression of tumors. The data from this study shed some light on the function, as well as the general network mechanisms of MASPIN in lung cancer.