Figure 8. The combined effect of overexpression of FAM83H and knock-down of SCRIB on the proliferation and tumor formation of gastric cancer cells. (A, B) The effect of overexpression of FAM83H and/or knock-down of SCRIB on the proliferation of MKN45 and NCI-N87 gastric cancer cells were evaluated with an MTT assay (A) and a colony-forming assay (B). The MTT assay was performed after seeding 3,000 cells per well of a 96-well plate. The colony-forming assay was performed by plating 1,000 cells per well in a 6-well plate for ten days. The number of colonies was determined with GeneTools analysis software. (C, D) In vivo tumor growth was evaluated by subcutaneously implanting 2x106 NCI-N87 cells with overexpression of FAM83H and/or knock-down of SCRIB. The tumor volume was measured every week after tumor implantation by the equation V = LxWxHx0.52 mm3 (C). At five weeks after tumor inoculation, the mice were euthanized and tumor weight was measured (D). (E) The mice were evaluated for metastasis and histologic findings of the lung. The arrows indicate metastatic NCI-N87 cells in lung. *; P < 0.05, **; P < 0.001.