The Golgi apparatus serves as a central hub for trafficking, regulating the anterograde transport and modification of proteins and lipids, or recycling extracellular proteins via retrograde transport through endosomes. Several Golgi-localized proteins have been reported to be involved in regulating cancer cell migration and invasion. Our previous study showed that the Golgi tethering factor golgin-97 is downregulated in breast cancer and is correlated with patient prognosis. Here, we demonstrated that the knockout of golgin-97 promotes tumor malignancy in breast cancer in vivo. We identified differentially expressed proteins between golgin-97 knockout cells and their parental breast cells using a quantitative proteomic approach based on 6-plex tandem mass tags (TMT) technology. Bioinformatics analysis further revealed that golgin-97 knockout was significantly related to inflammation-related factors and MAPK signaling pathways. We also integrated proteomic and genomic datasets to explore the biological impacts of golgin-97 in regulating breast cancer progression.