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. 2011 Aug 29;31(16):2062–2074. doi: 10.1038/onc.2011.383

Figure 2.

Figure 2

Reconstitution of miR-200 levels inhibits cellular migration, tumor growth and metastasis formation. (a) Stable transfection of pri-miR-200ba429, pri-miR-200c141 and both clusters (pri-miR-200ba429 and -200c141) in the CpG island-hypermethylated cancer cell lines RKO caused decreased migration capability revealed by wound-healing assay. Accurate measures of the wounds were taken at 0, 24, 48 and 72 h to calculate the migration rate. Data are means±s.d. from two independent experiments (Fisher's exact test P<0.01 vs empty vector in all experiments). Illustrative examples of the time-coursed wound-healing assays are shown in the upper panel. (b) Tumor development of xenograft mouse models was monitored during 45 days after subcutaneous injection of empty vector or pri-miR-200c141 stably transfected RKO cells. Restoring of miR-200c141 in the hypermethylated RKO cells led to a significant reduction of tumor volume (Student's t-test P=0.02) and weight (Student's t-test P=0.04). Representative tumors are shown in the upper panel. (c) Decreased metastatic potential by forced miR-200c141 expression in methylated RKO cells. The presence of metastases was evaluated after 2 months of intrasplenic injection of pri-miR-200c141 or empty-vector RKO-transfected cells. Illustrative examples are shown in the left panel. Metastases are indicated by arrows.