Figure 2

SPOP inhibits gastric cancer cell proliferation and migration. (A) The expression of SPOP protein in human GC cell lines - MKN45, MKN28, AGS, SGC7901, and human gastric mucosal cell line GES-1 by Western blotting. (B) Detection of SPOP expression in transfected AGS cells. Expression of SPOP was detected in cell lysates of SPOP-V5 transfected AGS cells by Western blotting. Control, AGS cells with empty vector plasmid; #1 and #2 represent different groups of stable AGS cell lines transfected with SPOP. (C) Decreased viability of gastric cancer cells transfected with SPOP. The viability of AGS cells was assessed by using an MTT assay. Data represent the average of three experiments (mean ± SD). **P < 0.01, compared to control cells. (D, E) Reduced migration of AGS cells following treatment with SPOP transfection. Cell migration was examined using a scratch assay. D, images acquired at indicated time points. E, quantification of cell migration rates. Data represent the average of three experiments (mean ± SD). **P < 0.01, compared to DMSO treated cells. (F, G) Colony formation of SPOP over-expressed AGS cells. One or two hundred stable cell lines with SPOP over-expression were seeded into 12-well plates for 2 weeks. Colonies were fixed and stained with crystal violet, then counted if the colonies > 50 cells. G, histograms show the rate of colony formation. Data represent the average of three experiments (mean ± SD). **P < 0.01, compared to DMSO treated cells.