Over the past four decades, the spreading of gastric adenocarcinoma has been increased statically world-wide. Even though cancer treatment is really challengeable as metastasis still shows critical stage for treatment in cancer patients. However, most of drugs have cytotoxic effect on cancer cells but failed at metastatic state. Maghemite nanoparticles encompass massive biomedical applications inducing cytotoxic effects. In this study, we focussed to evaluate the anti-metastatic and cell regulatory consequence of maghemite nanoparticles on gastric adenocarcinoma (AGS) cell. The anti metastatic strength was questioned by monitoring the production of metastatic factor nitric oxide (NO) which shows statistically considerable reduction while compared to untreated AGS cells. Clonogenic assay revealed low cloning efficiency and proliferation of these cells. Failing wound closer and cell migration, defeat of cell adhesion to extracellular matrix owing to decreased expression of cell adhesion factors in nanoparticle treated cells were noted from scratch assay analysis and cell adhesion assay respectively. Semi-quantitative PCR for cell cycle regulative genes illustrated that maghemite nanoparticles intention to arrest AGS cells at the G2/M phase transition and leads to progression of mitotic catastrophe. Further, analysis of proteins via two dimensional gel electrophoresis and LC-MS/MS endorsed the down regulation of proteins. The identified proteins are mainly concerned in the regulation of cell survival, proliferation, cell morphology and structure maintenance, drug resistance, tumour genesis and metastasis of AGS cells. From the results obtained here, it was concluded that maghemite nanoparticles have potent anti-metastatic activity and persuade cell death via mitotic catastrophe in gastric cancer cells.