Table 2.
The synthetic MMP inhibitor GM6001 decreases distance and reduces velocity of lipid-induced HT1080 motility in 3D collagen matrices but not on 2D collagen substrates.
| Condition (over 24 hours) | Avg. Distance μm (2D) | Avg. Distance μm (3D) | Avg. Velocity μm/min (2D) | Avg. Velocity μm/min (3D) |
| DMEM | 303.68 ± 71.1 | 73.07 ± 31.74 | 0.21 ± 0.049 | 0.050 ± 0.022 |
| DMEM + GM6001 | 277.38 ± 29.38 | 45.06 ± 15.17 | 0.19 ± 0.020 | 0.031 ± 0.010 |
| LPA | 298.58 ± 41.18 | 128.00 ± 56.98 | 0.21 ± 0.029 | 0.088 ± 0.039 |
| LPA + GM6001 | 303.18 ± 55.41 | 40.43 ± 6.08 | 0.21 ± 0.038 | 0.028 ± 0.0042 |
| S1P | 303.05 ± 33.78 | 116.24 ± 25.39 | 0.21 ± 0.028 | 0.080 ± 0.017 |
| S1P + GM6001 | 322.93 ± 69.05 | 44.598 ± 9.25 | 0.23 ± 0.049 | 0.031 ± 0.0057 |
| Luciferase* | N/A | 138.43 ± 57.71 | N/A | 0.096 ± 0.040 |
| si MT1-MMP* | N/A | 92.15 ± 35.90 | N/A | 0.064 ± 0.025 |
Nuc-GFP HT1080 cells were seeded on 2D plastic dishes coated with 50 μg/ml of collagen or embedded within a 3.75 mg/ml type I collagen matrix and their motility was analyzed using Metamorph® software (see Fig. 7) in the presence of the specified lipid with or without GM6001. siRNA (Luciferase or custom MT1-MMP) treated nuc-GFP HT1080 cells were only analyzed in 3D (see Fig. 14). Data are expressed as the average total distance (μm) or average velocity (μm/min) ± S.D. of 15 tracings per condition. [Note: 96 distances and velocities (144 for siRNA experiments) were generated per tracing for a total of 1440 (2160) data points]. Statistical significance of means was determined using either One-way ANOVA or t-test where p < 0.05 was deemed statistically signficant (see text for specific comparisons).