HER-2 overexpression modulates transforming growth factor (TGF)-β responses in a cell context dependent manner. (a-c) HER-2 overexpression in MCF-7 breast cancer cells blocks TGF-β mediated growth arrest. CN and H2 cells were treated with diluent control (diamonds) or TGF-β1 (0.2 ng/ml (black circles); 0.4 ng/ml (white circles); or 0.8 ng/ml (triangles)) and counted on the indicated days after treatment. Each point is the average of triplicate wells (± standard deviation for error bars). (a) The growth of MCF-7 CN cells is severely inhibited by TGF-β1. (b) MCF-7 H2 cells are resistant to growth inhibition by TGF-β1. (c) MCF-7 CN (diamonds) versus MCF-7 H2 (white circles) cells treated with 0.2 ng/ml TGF-β1. (d) ZR-75-1 cells are resistant to growth inhibition by TGF-β1 without HER-2 overexpression (ZR-75-1/CN (diamonds) versus ZR-75-1/H2 (white circles)) treated with 0.2 ng/ml TGF-β1. (e) TGF-β1 stimulates the growth of MDA-MB-231 H2 cells. MDA MB-231 CN (diamonds) and H2 cells (white cirlcles) were grown for 6 days in the presence of TGF-β1, β2 or β3 (0.01 to 100 ng/ml) or a diluent control. Cells were pulsed with [3 H] thymidine for the final 24 h of assay and thymidine incorporation was measured. The average counts of triplicate wells for each data point are represented as % of diluent control. (f) TGF-β induces a 'piling' phenotype in MDA-MB-231 that is dependent on HER-2 overexpression. MDA-MB-231 CN and H2 cells were grown for 5 days in the presence of 10 ng/ml TGF-β1 or diluent control. Cells were stained with crystal violet dye and photographed with a 20× (top four panels) or a 60× (bottom two panels) objective.