Figure 6

Over-expression of miR-145 in SW620 cell line affects cell morphology and proliferation. (A) The genomic region surrounding the miR-145 gene was PCR-amplified and cloned into pSilencer 4.1 under control of the CMV promoter. Mature miR-145 was detected by Northern analysis in a pooled population of SW620 cells following transfection. U6 snRNA was used as a loading control. (B) A major distinguishing feature of the cell population over-expressing miR-145 was the change in cell morphology from the round single cells of SW620 to elongated cells with extended processes typical of fibroblast-like cells. (C) The miR-145-expressing SW620 cell population showed a two-fold increase in anchorage-independent growth when grown in the presence of serum and a greater than 50% increase in cell proliferation/metabolic activity when grown in the presence (solid bars) or absence (open bars) of serum. *** p < 0.001. (D) Western analysis of the SW620/miR-145 cells and control cells shows the steady-state levels of E-cadherin were 50% lower in SW620 cells expressing the mature miR-145. FBS = fetal bovine serum, SF = serum-free.