FIGURE 6.

Increased CD147 and MT1-MMP expression and invasiveness in MCF-10A-K-Ras^V12 cells is mediated by MEK-ERK signaling. A, Western blot depicting 10A-K-Ras^V12 cells treated with vehicle, U0126 (10 μm), or PD98059 (25 μm) for 12 h. β-Tubulin was used as a loading control; n = 3. B, CD147 mRNA levels of 10A-K-Ras^V12 cells treated with vehicle or UO126 (10 μm) as measured by quantitative PCR and determined by cycle threshold values (see “Experimental Procedures” for details); normalized to β-actin; n = 3; ns, not significant. C, quantitation of cell invasion through Matrigel by 10A-K-Ras^V12 cells pretreated with vehicle, U0126 (10 μm), or PD98059 (25 μm) for 24 h. Columns, mean number of invasive cells/field ± S.E. (error bars); n = 3; *, p < 0.05; **, p < 0.01. D, representative micrographs showing invadopodia in 10A-K-Ras^V12 cells pretreated for 30 min with vehicle, U0126 (10 μm), or PD98059 (25 μm) and then seeded on fluorescent gelatin matrix in the presence of vehicle or inhibitors; see “Experimental Procedures” for further details. Red arrowheads, examples of invadopodia. Scale bar, 10 μm. E, percentage of cells degrading underlying matrix; or F, quantitation of invadopodia/cell in 10A-K-Ras^V12 cells treated with vehicle, U0126 (10 μm), or PD980539 (25 μm). Each invadopodia parameter was calculated by evaluating random fields containing at least 15 cells/field over three independent experiments. Column values are means ± S.E.; ***, p < 0.001.