Table 2.
Measurement of HA-Mediated Breast Tumor Cell Invasion.
| (A) Effects of HA (LMW-HA vs. HMW-HA) on Breast Tumor Cell Invasion: | |
|---|---|
| Treatments | Tumor Cell Invasion (% of control) |
| No treatment (control) | 100 ± 5 |
| LMW-HA treatment | 289 ± 10a |
| LMW-HA + anti-CD44 antibody treatment | 102 ± 3a |
| LMW-HA + cytochalasin D treatment | 75 ± 2a |
| HMW-HA treatment | 108 ± 4a |
| HA fragment (2–3 disaccharide) treatment | 96 ± 2a |
| Normal IgG + no HA treatment | 102 ± 3a |
| Normal IgG + LMW-HA treatment | 279 ± 8a |
| (B) Effects of AFAP-110 siRNA on HA-mediated Breast Tumor Cell Invasion: | ||
|---|---|---|
| Treatments | Tumor Cell Invasion (% of control) |
|
| Scrambled siRNA-treated cells | AFAP-110 siRNA-treated cells | |
| No treatment (control) | 100 ± 4 | 88 ± 3b |
| LMW-HA treatment | 285 ± 8b | 82 ± 2b |
| HMW-HA treatment | 105 ± 5b | 84 ± 4b |
| HA fragment (2–3 disaccharide) treatment | 98 ± 3b | 86 ± 3b |
Twenty-four transwell units containing 5µm porosity polycarbonate filters coated with the reconstituted basement membrane substance Matrigel were used for monitoring in vitro tumor cell invasion. Cell invasion processes were determined by measuring the cells that migrated and invaded to the lower side of the polycarbonate filters by standard cell number counting methods. The CD44-specific cell invasion was determined by subtracting non-specific cell invasion [i.e., cell that migrated (invaded) to the lower chamber in the presence of anti-CD44 antibody treatment]. The CD44-specific cell invasion in untreated cells (Table 2-control) or scrambled siRNA-treated cells without HA (Table 2B-control) is designated as 100%. All data represent mean ±SEM of the number of cells undergoing invasion detected in each sample. The value represents an average of triplicate determinations of five experiments with an S.D. value less than ±5%.
Statistically significant (p<0.001; analysis of variance; n=5) as compared with control samples [e.g., untreated cells (control)].
Statistically significant (p<0.005; analysis of variance; n=5) as compared with control samples [e.g., scrambled siRNA-treated cells (control) without HA].