The effects of Y27632 and EGF on pancreatic cancer cell proliferation. (A) Panc1, KP3 and AsPc1 cells were pretreated with 3 μM Y27632 or vehicle in RPMI containing 0.3% FCS for 1 h, and then stimulated with 30 ng/ml of EGF or vehicle for 24 h. The BrdU incorporation as the % of the control (lane 1) is shown. (B) Panc1 cells were treated with the indicated doses of Y27632 or vehicle in RPMI containing 3% FCS for 48 h, and the cell viability assay was performed using the MTT cell proliferation kit I. (C) The attached cells were treated with 0.5 μg/ml of normal mouse-IgG (open circle) or anti-EGFR-neutralizing antibodies (closed circle) for the indicated periods in medium containing 3% FCS, and the surviving cells were counted using the MTT cell proliferation kit I. The results are expressed as the absorbance (OD 560 nm-OD 750 nm). All assays were done in triplicate. (*) indicates a significant difference (p < 0.05) compared with lane 1, (#) indicates a significant difference (p < 0.05), compared with lane 2.