Table 2.
Reversal of CM-induced resistance by inhibition and removal of FGF, and induction of resistance by recombinant FGF
| Concentration producing 50% inhibition
|
|||
|---|---|---|---|
| Paclitaxel, nM | Doxorubicin, nM | 5-Fluorouracil, μM | |
| Neutralizing mAb to reverse resistance | |||
| aFGF mAb | |||
| No CM, no mAb (control) | NM | 7.95 ± 0.45 | NM |
| No CM + 5 μg/ml bFGF mAb | NM | 7.64 ± 0.48 | NM |
| + CM, no mAb | NM | 75.4 ± 3.56* | NM |
| + CM + 5 μg/ml IgG | NM | 75.2 ± 3.78* | NM |
| + CM + aFGF mAb, 0.05 μg/ml | NM | 73.4 ± 5.85* | NM |
| + CM + aFGF mAb, 0.1 μg/ml | NM | 45.5 ± 3.16* | NM |
| + CM + aFGF mAb, 1 μg/ml | NM | 30.1 ± 4.05* | NM |
| + CM + aFGF mAb, 5 μg/ml | NM | 29.2 ± 2.98* | |
| bFGF mAb | |||
| No CM, no mAb (control) | 10.1 ± 0.39 | 8.04 ± 0.31 | 0.74 ± 0.05 |
| No CM + 5 μg/ml bFGF mAb | 9.84 ± 3.98 | 8.48 ± 1.15 | 0.75 ± 0.02 |
| + CM, no mAb | 35.7 ± 1.02* | 32.1 ± 0.82* | 1.89 ± 0.15* |
| + CM + bFGF mAb, 0.05 μg/ml | 28.5 ± 1.47* | 27.7 ± 3.61* | 1.62 ± 0.11* |
| + CM + bFGF mAb, 0.1 μg/ml | 21.1 ± 1.00* | 15.0 ± 0.80* | 1.16 ± 0.06* |
| + CM + bFGF mAb, 0.5 μg/ml | 18.0 ± 0.67* | 11.7 ± 1.08* | 1.11 ± 0.12* |
| + CM + bFGF mAb, 5 μg/ml | 14.5 ± 1.90 | 8.24 ± 1.81 | 0.97 ± 0.09 |
| Removal of FGF by immunoprecipitation plus reconstitution with 0.16 ng/ml r-aFGF and/or 0.9 ng/ml r-bFGF | |||
| No CM (control) | NM | 8.99 ± 0.63 | NM |
| + CM | NM | 78.1 ± 5.32* | NM |
| + CM, remove aFGF | NM | 30.6 ± 1.77* | NM |
| + CM, remove aFGF, add r-aFGF | NM | 77.3 ± 3.68* | NM |
| + CM, remove bFGF | NM | 8.22 ± 0.59 | NM |
| + CM, remove bFGF, add r-bFGF | NM | 74.6 ± 2.85* | NM |
| + CM, remove aFGF & bFGF | NM | 8.47 ± 0.91 | NM |
| + CM, remove aFGF & bFGF, add r-aFGF | NM | 8.52 ± 0.74 | NM |
| + CM, remove aFGF & bFGF, add r-bFGF | NM | 28.3 ± 1.93* | NM |
| + CM, remove aFGF & bFGF, add r-aFGF & r-bFGF | NM | 76.8 ± 3.95* | NM |
| Resistance induced by r-aFGF and r-bFGF, independent of CM | |||
| No CM (control) | 1.18 ± 0.04 | 8.05 ± 1.80 | 0.81 ± 0.04 |
| + CM | 3.82 ± 0.07* | 44.6 ± 1.92* | 3.22 ± 0.14* |
| + r-aFGF, 1 ng/ml | 1.15 ± 0.06 | 8.10 ± 0.27 | 0.82 ± 0.08 |
| + r-aFGF, 10 ng/ml | 1.14 ± 0.03 | 8.43 ± 0.24 | 0.70 ± 0.05 |
| + r-aFGF, 50 ng/ml | 1.18 ± 0.05 | 8.05 ± 0.40 | 0.73 ± 0.05 |
| + r-bFGF, 1 ng/ml | 1.21 ± 0.03 | 8.54 ± 2.69 | 0.85 ± 0.12 |
| + r-bFGF, 10 ng/ml | 2.06 ± 0.35 | 23.8 ± 2.21* | 2.41 ± 0.08* |
| + r-bFGF, 50 ng/ml | 3.27 ± 0.85* | 50.8 ± 2.46* | 4.30 ± 0.74* |
| + r-aFGF, 0.04 ng/ml + r-bFGF, 0.9 ng/ml | 1.15 ± 0.05 | 8.53 ± 0.51 | 0.76 ± 0.09 |
| + r-aFGF, 0.08 ng/ml + r-bFGF, 0.9 ng/ml | 2.13 ± 0.06* | 15.2 ± 0.52* | 1.15 ± 0.11* |
| + r-aFGF, 0.16 ng/ml + r-bFGF, 0.9 ng/ml | 3.14 ± 0.14* | 35.3 ± 1.11* | 1.64 ± 0.07* |
| + r-aFGF, 0.32 ng/ml + r-bFGF, 0.9 ng/ml | 4.68 ± 0.09* | 62.2 ± 3.49* | 2.54 ± 0.15* |
| + r-aFGF, 0.64 ng/ml + r-bFGF, 0.9 ng/ml | 5.79 ± 0.16* | 84.9 ± 6.31* | 3.77 ± 0.13* |
| + r-aFGF, 0.9 ng/ml + r-bFGF, 0.9 ng/ml | 6.86 ± 0.22* | 107 ± 2.94* | 5.74 ± 0.28* |
For the neutralizing mAb experiment, the source of CM was lung histocultures for the aFGF mAb experiment, and early monolayer lung cultures for the bFGF mAb experiment. For the immunoprecipitation plus reconstitution experiment, the source of CM was lung histoculture. Mean ± SD. Drug effect was measured by the BrdUrd incorporation method. NM, not measured. Similar results were obtained for rat MAT-LyLu cells and human PC3 cells. Results in the top and middle parts are for rat tumor cells, and results in the bottom part are for PC3 cells. *, P < 0.05 compared with control (one way ANOVA).