Tumor cell invasion of collagen matrices requires coordinate lipid agonist-induced G-protein and membrane-type matrix metalloproteinase-1-dependent signaling

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).

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