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miR-34a Inhibits Migration and Invasion of Tongue Squamous Cell Carcinoma via Targeting MMP9 and MMP14

Figure 2

miR-34a targets MMP9 and MMP14.

(A), The sequence of miR-34a (middle) matches the coding sequence (CD) of MMP9 and 3′untranslated region (UTR) of MMP14 (top). Bottom, mutations of the CD of MMP9 and 3′UTR of MMP14. (B), miR-34a inhibited wild-type, but not mutated MMP9 CD and MMP14 3′UTR luciferase reporter activity. CAL27 cells were co-transfected with firefly luciferase reporter plasmids containing wild type or mutant MMP9 CD and MMP14 3′UTR, and pRL-TK plasmid (a plasmid expressing rellina luciferase) and pcDNA3.0-miR-34a (miR-34a) or pcDNA3.0 (NC) as indicated. After 24 h, firefly luciferase activities were measured and normalized by use of renilla luciferase activities. Data were presented as mean ±SD (*P<0.05). (C), The relative MMP9 and MMP14 mRNA levels determined by qRT-PCR in miR-34a or NC transfected CAL27 cells (*P<0.01). (D), Inhibition of the expression of MMP9 and MMP14 proteins. Representative Western blotting image (top) and the quantification (bottom) of MMP9 and MMP14 proteins in miR-34a transfected SCC-15 and CAL27 cells. β-actin was used as internal control and was also detected by Western blot (**P<0.01).

Figure 2

doi: https://doi.org/10.1371/journal.pone.0108435.g002