miR-196a directly targets theHOXA5gene. (A) A human HOXA5 or FOXO1 3’ untranslated region (UTR) fragment containing the wild-type or mutant miR-196a binding sequence was cloned downstream of the luciferase reporter gene. (B) The luciferase reporter plasmid containing wild-type or mutant HOXA5 and FOXO1 3’UTR were co-transfected into HEK-293 T cells with pCDNA-miR-196a or pCDNA-miR-NC. Luciferase activity was determined using the dual luciferase assay and shown as the relative firefly activity normalized to renilla activity. (C) qRT-PCR analyses of HOXA5 mRNA level following treatment of SPCA1 cells with miR-196a inhibitors or A549 cells with miR-196a mimics. (D) Western blot analyses of HOXA5 protein levels following treatment of SPCA1 cells with miR-196a inhibitors or A549 cells with miR-196a mimics. GAPDH was used as control. *P < 0.05, **P < 0.01 and N.S. not significant.
Liu et al. BMC Cancer 2012 12:348 doi:10.1186/1471-2407-12-348