Effect of signal transduction pathways inhibition on cell migration of HCC827 GR5 cells deprived of gefitinib.

<div><p>(A) HCC827 GR5-G (maintained in the absence of gefitinib for 7 days) cells were exposed to dasatinib 0.01μM, SU11271 1μM, U0126 10μM, NVP-BEZ235 0.1μM, gefitinib 1μM or SU11271 1μM + gefitinib 1μM during migration time. Columns, means of 10 fields counted; bars, SD. Result is representative of three independent experiments. (B) HCC827 GR5-G cells were incubated with dasatinib 0.01μM, SU11271 1μM, gefitinib 1μM or SU11271 1μM + gefitinib 1μM. After 24h cells were counted and cell death was evaluated by fluorescence microscopy on Hoechst/PI stained cells. Columns, means of three independent experiments *, significance vs HCC827 GR5-G. ***P<0.001, **P<0.01; ^###P<0.001, ^##P<0.01.</p> <p>HCC827 GR5-G were transfected with Src, STAT5a/b, p38 siRNA or control siRNA (scramble) for 48 h. Then medium was replaced with fresh medium for 16h and the expression of the indicated proteins was analyzed by Western blotting (C) or cells were seeded on culture inserts for migration assay (D). Columns, means of 10 fields counted; bars, SD. Results are representative of three independent experiments. ***P<0.001. </p>