TABLE 1.
Incidences of mammary tumors, metastases, and invasion in rats injected with various transfected cell lines
| Transfected DNA (designation of cell line)a | Incidence of mammary tumorsb | p valuec | Incidence of metastasisd | p valuee | Incidence of invasionf | p valueg |
|---|---|---|---|---|---|---|
| % | % | % | ||||
| None; untransfected Rama 37h | 18/20 (90) | p = 0.99 | 0/20 (0) | p < 0.0001 | 0/20 (0) | p < 0.0001 |
| Vector onlyh | ||||||
| Clone | 19/20 (95) | p = 1.00 | 0/20 (0) | p < 0.0001 | ||
| Pool | 23/23 (100) | p = 1.00 | 2/23 (9) | p < 0.0001 | 3/18 (16) | p < 0.0001 |
| S100A4 wild typeh | ||||||
| Clone | 22/22 (100) | 20/22 (90) | ||||
| Pool | 19/19 (100) | 17/19 (89) | 17/20 (85) | |||
| S100A4 mutant 72 | ||||||
| Clone | 16/16 (100) | p = 1.00 | 7/16 (43) | p = 0.0029 | ||
| Pool | 19/19 (100) | p = 1.00 | 11/19 (57) | p = 0.063 | 9/19 (47) | p = 0.017 |
| S100A4 mutant 72.78 | ||||||
| Clone | 18/19 (94) | p = 1.00 | 3/19 (15) | p < 0.0001 | ||
| Pool | 21/23 (91) | p = 1.00 | 2/23 (8) | p < 0.0001 | 3/5 (60) | p = 0.289 |
| S100A4 mutant 16.72.78 | ||||||
| Clone | 16/16 (100) | p = 1.00 | 0/16 (0) | p < 0.0001 | ||
| Pool | 14/15 (93) | p = 1.00 | 6/31 (19) | p < 0.0001 | 7/17 (41) | p = 0.007 |
a S100A4 mutant 72, cells expressing mutant S100A4 protein in which residue 72 has been changed from phenylalanine to glutamine; S100A4 mutant 72.78, cells expressing mutant S100A4 protein in which residues 72 and 78 have been changed from phenylalanine to glutamine and alanine respectively; S100A4 mutant 16.72.78, cells expressing mutant S100A4 protein in which residues 16, 72, and 78 have been changed from phenylalanine to alanine, glutamine, and alanine, respectively.
b Number of tumors/number of animals inoculated.
c p values for tumor incidences in cloned or in pooled transfectant cells of mutant 72, mutant 72.78, and mutant 16.72.78 are compared with cloned or pooled transfectant cells of the wild type S100A4 transfections (Fisher's Exact test).
d Numbers of animals (%) with lung metastases/numbers of animals with tumors.
e p values for incidences of metastasis in cloned cell lines or in cell pools transfected with S100A4 mutant 72, mutant 72.78, mutant 16.72.78, empty pBKCMV vector, or cloned untransfected Rama 37 cells compared with cloned or pooled transfectant cells of the wild type S100A4 transfections (Fisher's Exact test).
f Numbers of animals (%) with muscle invasion/numbers of animals with muscle around tumors. No peripheral muscle tissue was recovered in sections of primary tumors from 10 rats with clones and 11 rats with pools of wild type S100A4, from 7 rats with clones and 9 rats with pools of mutant 72, in 16 rats with clones and 21 rats with pools of mutant 72.78, in 8 rats with clones and 22 rats with pools of mutant 16.72.78, and in 11 rats with clones and 14 rats with pools of pBKCMV vector transfected cells. In view of the low proportion of primary tumors with muscle in the available histological sections of wild type, mutants, and vector control, the results for clones and pools were combined for statistical analysis.
g p values for muscle invasion in tumors from combined cell lines and pooled transfectant cells of mutant 72, mutant 72.78, and mutant 16.72.78, pBKCMV vector or cloned untransfected Rama 37 cells compared with combined cloned and pooled transfectant cells of the wild type S100A4 transfectants (Fisher's Exact test).