Objective To investigate the effect of RNAi silencing c-Met gene on the proliferation and invasion ability of human glioma U251 cell. Methods Three kinds of short hairpin RNA (shRNA) targeting c-Met gene were constructed. All of the shRNAs were transfected into U251 cell by liposome transfection, in order to screen out the stable expression cell line. The expression of c-Met in U251 cell was detected by PCR and Western Blot, in order to screen out the effective shRNA. The proliferation of U251 cell was detected by MTT assay and flow cytometry. The invasion and migration of U251 cell was detected by transwell assay and cell scratch. Results The shRNA targeting c-Met gene was successful constructed. The expression of c-Met was significantly decreased after c-Met gene was silenced, the difference was statistically significant (P < 0.05). The proliferation, invasion, migration and tumorigenicity of U251 cell was inhibited after silencing c-Met gene, the difference was statistically significant (P < 0.05). Conclusions RNAi silencing c-Met gene can effectively inhibit the proliferation, invasion, migration and tumorigenicity of human glioma U251 cell.