Objective To evaluate the effects of gemcitabine combined with sorafenib on the proliferation, apoptosis, migration, and invasion of gallbladder cancer cell line SGC996. Methods The cells were treated with gemcitabine and/or sorafenib. The concentrations of the group treated with gemcitabine were 0.1, 1.0, 2.0, 4.0, and 10.0 μg/mL. The concentrations of the group treated with sorafenib were 0.1, 1.0, 2.5, 5.0, 10.0, and 20.0 μmol/L. The concentrations of the group treated with gemcitabine and sorafenib were as follows: gemcitabine (2.0, 4.0, and 10.0 μg/mL) and sorafenib (2.5, 5.0, 10.0, and 20.0 μmol/L). The cells of control group were not treated with gemcitabine or sorafenib. The proliferation of SGC996 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The migration and invasion of SGC996 cells were examined using Transwell assay. The apoptosis of SGC996 cells was examined by FITC-Annexin V/PI double staining. Results Sorafenib inhibited the proliferation of SGC996 cells, and this growth inhibition was dependent on time and dose (P<0.05). Sorafenib also inhibited migration and invasion of SGC996 cells (P<0.05), but it did not induce apoptosis (P>0.05). Gemcitabine induced apoptosis and inhibited invasion of SGC996 cells (P<0.05), but it did not inhibit proliferation and migration of SGC996 cells (P>0.05).The combination of gemcitabine and sorafenib induced apoptosis of SGC996 cells and significantly inhibited the proliferation, migration, and invasion of SGC996 cells (P<0.05). Conclusion The antitumor ability of combination of gemcitabine and sorafenib is much higher than that of either gemcitabine or sorafenib, which may be an effective chemical therapy for gallbladder cancer.