Figure 5

Over-expression of miR-143 in the SW620 cell line affects cell morphology and proliferation. (A) MiR-143 genomic DNA was cloned under control of the U6 promoter (in pSilencer 2.1) and transfected into SW620 cells. Seven stable SW620 clones were identified that expressed miR-143 (A4, B3, B5, B6, C1, C5, and D2). U6 snRNA was used as a loading control. (B) The seven SW620/miR-143 clones were examined for cell morphology compared to vector control. (C) Western analysis of the SW620/miR-143 clones and control cells shows the steady-state levels of E-cadherin. E-cadherin ratios were calculated using β-actin as a normalization control. (D) Seven SW620/miR-143 clones were assayed for proliferation/metabolic activity compared with vector control when grown in the absence (open bars) or presence of serum (solid bars). (E) The same clones were examined for anchorage-independent cell growth in the rapid soft agar assay in the presence of serum. Two independent experiments were performed. ** p < 0.01, *** p < 0.001.