Fig. 3. Down-regulation of miR-184 enhances tumorigenic hallmarks of human SCC.

A In situ hybridization of miR-184 shows high signal in suprabasal cells of the human skin. B RNA was extracted from human skin SCC C12C20 cells or from primary human foreskin cells, as a control (healthy). TaqMan real time PCR assay of miR-184 shows a lower level of miR-184 in SCC. C–G Human C12C20 SCC cells were stably infected with lenti virus that carries an anti-miR-184 specific antagonist (SCC/AM) or an empty vector as a control (SCC/Ctl) (validation in Fig. S2). C Colony formation assay was performed for the indicated cells, showing increased colony number and size in SCC/AM, compared to controls. Lower panels show magnification of the marked areas. Quantification of average colony number is shown in (D) and average colony size is shown in (E). F Boyden chamber migration assay demonstrating higher migration capacity of AM infected cells compared to controls. G is quantification of (F) (as detailed in Methods). Data represents average and standard deviation from 3 biological replicates. Scale bars were 20 μm. Statistical significance was assessed by t test (*p < 0.05). H Tumor volume of C12C2O SCC cells expressing anti-miR184 (AM184) or control vector (Ctl) injected subcutaneously to nude mice (n = 5 per group; ****p < 0.0001).