ITGB1 (Integrin β1, CD29) is a member of the integrin family and roles as a major adhesion receptor. Gastric cancer (GC) is an important cause of mortality worldwide, especially in China. As a potential cancer enhancer, the roles ITGB1 plays in GC progression keep unclear. In the current study, our assay on the databases about tumor associated gene expression and interaction found that the high expression of ITGB1 was closely correlated with the poor prognosis of GC patients. To explore the roles ITGB1 plays in GC progression, an ITGB1 deleted cell line (ITGB1-/-SGC7901) was generated using CRISPR/Cas9 method. The tumor malignancy associated cell behaviors and microstructures were detected, imaged, and analyzed using MTT, wound healing, transwell, scanning electron microscope (SEM), laser scanning confocal microscope (LSCM) and others. The results indicated that ITGB1 deletion decreased the GC cell proliferation and motility, and inhibited motility relevant microstructures, such as pseudopodia and filopodia markedly in ITGB1deleted SGC7901 cells. The analysis of STRING database suggested that ITGB1 contributes to the malignancy of GC mediated by Src-mediated FAK/PI3K/Akt signaling pathways. Taken together, the results above showed that ITGB1 may be a potential targeting marker for GC diagnosis and therapy, and it enhances GC progression via Src-mediated FAK/PI3K/Akt signal pathway that needs further validation in future.