Figure 4.

Overexpression of DUSP8 Suppressed the Proliferation and Migration of Human Colorectal Carcinoma Cells In Vitro

Human CRC SW620 cells were transiently transfected with p-DUSP8 (2.5 μg) or p-EGFP (2.5 μg) in vitro. Forty-eight hours later, (A) the relative expression of DUSP8 was determined using real-time PCR, (B) the protein level of DUSP8 was examined using immunofluorescence, and (C) the proliferation of cells was detected using the CCK-8 assay. (D) The colony capability of a single cell was observed using microscopy (magnification 100×) and the colony area (square millimeters) was calculated using ImageJ software. (E) Colon formation assay also was performed and calculated at the indicated time point. (F) Migration of SW620 cells was performed using wound-healing assay (magnification 100×), and the open wound area (square millimeters) also was calculated using ImageJ software. (G) Expression of indicated cyclin-dependent kinases and metastasis-related molecules was detected using real-time PCR. (H) Protein expression of DUSP8, ERK, p-ERK, AKT, and p-AKT also was detected by western blot, and the gray intensity value was calculated using ImageJ software. Representative data from three independent experiments are shown. *p < 0.05 and **p < 0.01.