Article Figures & Data
Figures
- Fig. 1.
Fas and Fas-L expression. 100 μg of cellular extract was used for Western blotting analysis as described in “Materials and Methods.” Lane 1: SaOs-2; Lane 2: HT-1080; Lane 3: HS-16; Lane 4: HS-30; Lane 5: HS-42; Lane 6: A204; and Lane 7: M9110.
- Fig. 2.
Effect of CH-11 antibody on cytotoxicity-induced by anticancer drugs. Cells were exposed to anticancer drugs at different concentrations for 3 days or pretreated with CH-11 (6 ng/ml) for 1 h followed by anticancer drugs for 3 days. Cytotoxicity of drugs was determined using the SRB assay. A, HT-1080; B, HS-16; and C, WI-38. (○), MTX; (•), MTX + CH-11; (▵), DOX; (▴), DOX + CH-11; (□), paclitaxel; and (▪), paclitaxel + CH-11.
- Fig. 3.
Effect of CH-11 on induction of cell death by DOX and paclitaxel in HT-1080 cells. Cells were pretreated with CH-11 (6 ng/ml) for 1 h followed by DOX (0.5 μm) or paclitaxel (0.1 μm) for 24 h. Subdiploid cells were measured by FACS analysis as described in “Materials and Methods.” (□), untreated control; ([cjs2106]), CH-11; ([cjs2113]), DOX; (▪), DOX + CH-11; ([cjs2088]), paclitaxel; and ([cjs2110]), paclitaxel + CH-11. The figure is the representative of two similar experiments.
- Fig. 4.
Effect of CH-11 antibody on the expression of caspase-8 and caspase-9 and cytochrome c and activation of caspase-8. HT-1080 cells were treated with CH-11(6 ng/ml) or pretreated with SB203580 (10 μm) or z-IETD (20 μm), respectively, for 16 h followed by CH-11. Whole cell extracts (for caspase-8 and caspase-9) or cytosolic extracts (for cytochrome c) were prepared and used for Western blotting analysis. For caspase-8 activation assay, cells were lysed in hypotonic buffer as described in “ Materials and Methods.” A, expression of caspase-8 and caspase-9 and cytochrome c. Lane 1: untreated control; Lane 2: CH-11, 4 h; Lane 3: CH-11, 8 h; Lane 4: CH-11, 24 h; Lane 5: IETD(Ome)-fmk + CH-11, 8 h; Lane 6: IETD(Ome)-fmk + CH-11, 24 h; Lane 7: SB203580 + CH-11, 8 h; and Lane 8: SB203580 + CH-11, 24 h. B, activation of caspase-8. (○), untreated control; (•), CH-11, 1 h; (▵), CH-11, 4 h; (▿), CH-11, 8 h; and (▾), CH-11, 24 h.
- Fig. 5.
Effect of CH-11 on cell cycle distribution in response to anticancer drugs in HT-1080 cells. A, cells were incubated with CH-11 (6 ng/ml) for 24 or 48 h. B, cells were exposed to DOX (20 nm) or paclitaxel (2 nm) or to CH-11 (6 ng/ml) for 1 h followed by DOX (20 nm) or paclitaxel (2 nm) for 24 h. Cells were then harvested and subjected to FACS analysis.
- Fig. 6.
Activation of MAPKs by CH-11 and anticancer drugs and the effect of SB203580 on CH-11-induced activation of MAPKs and cytotoxicity of CH-11 plus anticancer drugs. A, activation of MAPKs by CH-11 and anticancer drugs. HT-1080 cells were exposed to CH-11(6 ng/ml), MTX (200 nm), DOX (20 nm), or paclitaxel (2 nm) at the indicated times, whole cell extracts were prepared for Western blotting analysis. Lane 1: untreated control; Lane 2: CH-11, 30 min; Lane 3: CH-11, 1 h; Lane 4: CH-11, 4 h; Lane 5: MTX, 1 h; Lane 6: DOX, 1 h; and Lane 7: paclitaxel, 1 h. B, the effect of SB203580 on CH-11-induced activation of MAPKs. HT-1080 cells were pretreated with SB203580 (10 μm) for 16 h and followed by CH-11(6 ng/ml) for an additional 4 h. Whole cell extracts were prepared for Western blotting analysis. Lane 1: untreated control; Lane 2: CH-11 alone; and Lane 3: CH-11 + SB203580. C and D, SB203580 inhibits the enhanced effect of CH-11 combined with anticancer drugs. Cells were pretreated with SB203580 (10 μm) or IETD(Ome)-fmk (20 μm) for 16 h then treated with CH-11 (6 ng/ml) for 1 h followed by MTX or DOX or paclitaxel for 3 days. Cell killing was determined using a SRB assay. (□), drug alone; ([cjs2113]), drug + CH-11; (), drug + CH-11 and SB203580; (▪), drug + CH-11 and IETD(Ome)-fmk; and ([cjs2106]), CH-11 alone. Drug concentration used in HT-1080 cells. (C): MTX, 200 nm; DOX, 20 nm; paclitaxel, 2 nm; and in HS-16 cells. (D): MTX, 500 nm; DOX, 100 nm; and paclitaxel, 10 nm.
Volume 1, Issue 14
