Introduction
Materials and methods
Primary cultures, ascites samples and human subjects
Reagents
Cell viability assays
Immunoblot analysis
Statistical analysis
Results
Effect of ascites on TRAIL sensitivity
Protein concentration in ascites and serum CA125 levels
Effect of ascites on drug sensitivity
Ovarian cancer ascites | Cisplatin IC50 (ng/ml) | Paclitaxel IC50 (ng/ml) | Doxorubicin IC50 (ng/ml) | Etoposide IC50 (ng/ml) | Vinorelbine IC50 (ng/ml) | TRAIL IC50 (ng/ml) | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
fluids | - | + | - | + | - | + | - | + | - | + | - | + |
346
| 746 ± 38 | 1400 ± 24 | 16 ± 7 | 100 ± 9 | 86 ± 8 | 250 ± 15 | 2183 ± 147 | 7500 ± 245 | 4.1 ± 0.25 | 10 ± 1 | 8.4 ± 2.7 | 28.6 ± 4 |
432
| 746 ± 38 | 900 ± 43 | 16 ± 7 | 16 ± 3 | 86 ± 8 | 70 ± 10 | 2183 ± 147 | 2000 ± 87 | 4.1 ± 0.25 | 4.4 ± 0.4 | 8.4 ± 2.7 | 8.1 ± 2.8 |
488
| 746 ± 38 | 900 ± 23 | 16 ± 7 | 37 ± 5 | 86 ± 8 | 107 ± 13 | 2183 ± 147 | 6000 ± 184 | 4.1 ± 0.25 | 6.3 ± 1 | 8.4 ± 2.7 | 9.0 ± 3 |
508
| 746 ± 38 | 2300 ± 16 | 16 ± 7 | 16 ± 4 | 86 ± 8 | 145 ± 5 | 2183 ± 147 | >50000 | 4.1 ± 0.25 | >1000 | 8.4 ± 2.7 | 38 ± 4.2 |
509
| 746 ± 38 | 3000 ± 54 | 16 ± 7 | 80 ± 3 | 86 ± 8 | 750 ± 8 | 2183 ± 147 | >50000 | 4.1 ± 0.25 | >1000 | 8.4 ± 2.7 | 32 ± 3.1 |
551
| 746 ± 38 | 820 ± 47 | 16 ± 7 | 13 ± 4 | 86 ± 8 | 112 ± 12 | 2183 ± 147 | 4600 ± 231 | 4.1 ± 0.25 | 6.6 ± 0.2 | 8.4 ± 2.7 | 15 ± 5.4 |
Ascites decrease TRAIL cytotoxicity in primary cultures of ovarian tumor cells and activate Akt in these cells
Primary samples | ascites OVC346 | ascites OVC509 | ||
---|---|---|---|---|
ascites | - | + | - | + |
218A
| 7.2 ± 1.3 | 12 ± 1.1 | 7 ± 0.7 | 14.5 ± 0.8 |
231A
| 4.8 ± 0.7 | 5.5 ± 0.7 | 4.4 ± 0.8 | 7.5 ± 0.6 |
238A
| 16 ± 1.8 | 19.5 ± 0.9 | 15 ± 0.4 | > 30 |
285A
| 12 ± 2.1 | 18.8 ± 1.4 | 14 ± 1.1 | > 30 |
318A
| 5.5 ± 0.5 | 7.5 ± 0.7 | 5.7 ± 0.6 | 12 ± 0.9 |
327A
| 5.3 ± 0.7 | 7.5 ± 0.5 | 5.2 ± 0.7 | 9.7 ± 0.8 |
339A
| 7.8 ± 1.1 | 14.5 ± 0.7 | 7.5 ± 1.2 | 19.5 ± 0.6 |
341T
| 10.8 ± 1.3 | 16.5 ± 1.2 | 10 ± 0.4 | 19.5 ± 0.9 |
346A
| 6.5 ± 0.9 | 9.2 ± 0.6 | 6.5 ± 0.6 | 16 ± 1 |
Prosurvival activity of ovarian cancer ascites and disease-free intervals
Characteristics | Non-protective ascites n = 17 | Protective ascites n = 18 |
P
|
---|---|---|---|
Age (yrs, median) | 62 (27-85) | 57 (36-88) | NS |
Histopathology | |||
Serous | 12 (71%) | 15 (83%) | NS |
Other | 5 (29%) | 3 (17%) | |
Grade | |||
1-2 | 10 (59%) | 12 (67%) | NS |
3 | 7 (41%) | 6 (33%) | |
Stage | |||
I or II | 4 (24%) | 3 (17%) | NS |
III or IV | 13 (76%) | 15 (83%) | |
Optimal surgery | |||
Yes | 9 (53%) | 10 (56%) | NS |
No | 8 (47%) | 8 (44%) |