Effect of Thrombin and TRAP of the PAR-1 Thrombin Receptor on Pulmonary Metastasis of B16F10 Melanoma
| Group . | Median No. of Nodules (range) . | Median Nodule Volume (μL; range) . | Tumor Mass (μL) . | Fold Increase . | P . |
|---|---|---|---|---|---|
| Experiment 1 (n = 12) | |||||
| Control | 1.5 (0-7) | 0.6 (0-6) | 0.9 | ||
| Thrombin | 7 (2-10) | 5.7 (1-15) | 40.0 | 44 | .02 |
| 100 μmol/L 13-mer control peptide | 2.5 (0-9) | 2.3 (0-15) | 5.8 | NS | |
| 100 μmol/L 14-mer TRAP | 5.5 (0-14) | 7.9 (0-39) | 43.5 | 48 (8)§ | .02 (0.03)* |
| Experiment 2 (n = 5) | |||||
| SALLRN | 7 (2-14) | 19.9 (2-59) | 139 | ||
| SFLLRN | 26 (13-31) | 219 (131-353) | 5,694 | 41 | .04 |
| Group . | Median No. of Nodules (range) . | Median Nodule Volume (μL; range) . | Tumor Mass (μL) . | Fold Increase . | P . |
|---|---|---|---|---|---|
| Experiment 1 (n = 12) | |||||
| Control | 1.5 (0-7) | 0.6 (0-6) | 0.9 | ||
| Thrombin | 7 (2-10) | 5.7 (1-15) | 40.0 | 44 | .02 |
| 100 μmol/L 13-mer control peptide | 2.5 (0-9) | 2.3 (0-15) | 5.8 | NS | |
| 100 μmol/L 14-mer TRAP | 5.5 (0-14) | 7.9 (0-39) | 43.5 | 48 (8)§ | .02 (0.03)* |
| Experiment 2 (n = 5) | |||||
| SALLRN | 7 (2-14) | 19.9 (2-59) | 139 | ||
| SFLLRN | 26 (13-31) | 219 (131-353) | 5,694 | 41 | .04 |
TRAP refers to 14 mer-SFLLRNPNDKYEPF or 6 mer-SFLLRN. Fifty thousand viable B16F10 cells were treated with either buffer, 1 U mL thrombin, 100 μmol/L TRAP, or 100 μmol/L control peptides, 13 mer SDEALPLGSPRCD, or 6 mer SALLRN for 60 minutes at 37°C. Cells were then washed three times with PBS-Ca-Mg-0.1 BSA before intravenous injection into the tail vein of syngeneic mice. Pulmonary metastases were examined macroscopically on day 21 for experiment 1 and on day 35 for experiment 2. n refers to number of animals in each subset of the experiment. P value refers to significance of the fold increase in tumor mass compared with control, determined by Wilcoxon Rank Sum test. The SEM is given.
Abbreviation: NS, not significant.
The number in parentheses refers to the increase over control peptide, with P value.