STAT3isactivatedinlungcancercellswithmutantEGFR.A,.pptVIP

STAT3 is activated in lung cancer cells with mutant EGFR. A, cells were starved in 0.5% serum overnight and left unstimulated or stimulated with 50 ng/mL EGF or treated with 1 μmol/L gefitinib for 2 or 24 hours. James V. Alvarez et al. Cancer Res 2006;66:3162-3168 ?2006 by American Association for Cancer Research STAT3 is activated in lung cancer cells with mutant EGFR. A, cells were starved in 0.5% serum overnight and left unstimulated or stimulated with 50 ng/mL EGF or treated with 1 μmol/L gefitinib for 2 or 24 hours. STAT3 tyrosine phosphorylation was determined by Western blotting. *, phosphorylated EGFR. B, cells were treated as in A , and STAT3 DNA-binding activity was measured by EMSA using a hSIE probe. The STAT3-DNA complex could be disrupted with an antibody against STAT3. Arrow, STAT3-DNA–binding complex in H3255 cells; arrowhead, STAT3-DNA complex in HCC-827 cells consistent with a STAT3:STAT1 heterodimer. C, cells were starved in 0.5% serum overnight and then treated with 1 μmol/L gefitinib or 20 μmol/L PD98059 for 2 hours, and STAT3 serine phosphorylation and ERK1/2 phosphorylation were determined by Western blotting. D, cells were transfected with a STAT3-dependent luciferase reporter m67-luc and treated with 20 μmol/L PD98059 or 10 μmol/L U0126 for 24 hours. Levels of STAT3-dependent luciferase production were measured and normalized to Renilla luciferase phRL-luc .