Figure 3

Repression of SPOP promotes proliferation and migration of gastric cancer cells. (A) MKN45 cells were transfected with GFP-tagged miR-SPOP and screened by flow cytometry for the stable cells. Western blotting was performed to detect SPOP expression. (B) Repression of SPOP enhances gastric cancer cell viability. The viability of MKN45 cells was assessed using an MTT assay. Data represent the average of three experiments (mean ± SD). *P < 0.05, compared to control cells. (C) Repression of SPOP promotes Ki-67 expression in nucleus. Immunofluorescent stainings of transfected miR-SPOP and endogenous Ki-67 were performed in MKN45 cells. MKN45 cells were transfected with GFP-tagged miR-SPOP for 48 h. Ki-67 was detected by incubating cells with mouse anti-Ki-67 and subsequently with Alexa Fluor 594 goat anti-mouse antibodies. Nucleus was identified by DAPI staining. (D, E) Repression of SPOP accelerates MKN45 cell migration in a scratch assay. Bars represent the standard deviation of three independent experiments. **P < 0.01, compared to DMSO treated control. (F) Repression of SPOP increases gastric cancer cell colony formation. Two hundred stable miR-SPOP transfected MKN45 cells were seeded into 12-well plates for 2 weeks. Colonies were fixed and stained with crystal violet and then counted if the colonies > 50 cells. (G) Histograms show the colony formation rate. Data represent the average of three experiments (mean ± SD). **P < 0.01, compared to DMSO treated cells.