Abstract: Objective To study the effects of glutamine synthetase (GS) on proliferation,migration and invasion of C6 stable cell lines which transfected with GS expression vector or shRNA vector. Methods C6 stable cell lines were generated by introducing GS shRNA,GS-EGFP-N3 or EGFP-N3 vector and selected with G418 followed by cloning.GS expression was detected by Western blot.Cell proliferation was detected by MTS and colony-forming experiment.Adhesion assay,wound healing and transwell experiments was used to detect the cell adhesion,migration and invasion. Results Western blot showed that GS was over-expressed in GS stable cells and no signal was detected in GS knocking-down cells.GS over-expression suppressed cell growth,clone formation and invasion,increased cell adhesion;GS knocking-down increased cell invasion,decreased cell adhesion,and had no significant impact on cell growth and clone formation. Conclusion The level of GS affects cell growth,migration and invasion of C6 glioma cells.