Effects of Proteasome Inhibitor on DNA Fragmentation in CLL Patient Isolates
| . | Patient Number . | Control* . | MPS . | MG132 . |
|---|---|---|---|---|
| Spontaneously apoptotic isolates | ||||
| 156 | 46.1 | 60.5 | 87.3 | |
| 160 | 27.4 | 34.1 | 82.7 | |
| 180 | 89.8 | 90.1 | 96.1 | |
| 213 | 24.9 | 48.5 | 70.1 | |
| 227 | 37.9 | 49.8 | 93.8 | |
| 228 | 51.2 | 71.2 | 97.3 | |
| 230 | 32.4 | 57.7 | 90.4 | |
| 232 | 25.6 | 37.8 | 91.2 | |
| 233 | 29.5 | 31.8 | 72.8 | |
| 235 | 38.5 | 57.6 | 89.9 | |
| Mean (SD) | 40 (19) | 54 (18) | 87 (9) | |
| Glucocorticoidsensitive isolates | ||||
| 131 | 0.7 | 66.3 | 99.5 | |
| 132 | 3.7 | 51 | 95.4 | |
| 134 | 8.9 | 23.4 | 99.6 | |
| 135 | 15.7 | 90.6 | 88.8 | |
| 137 | 11.2 | 37.9 | 88.6 | |
| 140 | 2.4 | 24.3 | 90.4 | |
| 144 | 4.6 | 34.1 | 50.9 | |
| 146 | 13.4 | 31 | 78.5 | |
| 147 | 1.4 | 19.6 | 92.5 | |
| 148 | 10 | 49.4 | 95.2 | |
| 149 | 1.6 | 30.1 | 99 | |
| 154 | 4.1 | 99.4 | 98.3 | |
| 155 | 16.7 | 36.9 | 99.8 | |
| 157 | 8.7 | 33.3 | 55.3 | |
| 188 | 3.3 | 33.3 | 36.8 | |
| 209 | 8 | 34.8 | 95.7 | |
| 210 | 3.7 | 37.1 | 88.9 | |
| 211 | 8.7 | 48.9 | 98.6 | |
| 212 | 13.8 | 41.8 | 89.3 | |
| 214 | 3.1 | 20.4 | 92.9 | |
| 218 | 2.3 | 23.5 | 96.7 | |
| 220 | 7.6 | 30.3 | 95 | |
| 222 | 2.5 | 30.5 | 80.1 | |
| 229 | 10.7 | 44.9 | 77.2 | |
| 238 | 5.2 | 31.7 | 92.2 | |
| 247 | 3 | 44.8 | 95.7 | |
| 250 | 18.5 | 33.2 | 82.7 | |
| 251 | 16.1 | 61.6 | 82.9 | |
| Mean (SD) | 7 (5) | 41 (19) | 87 (15) | |
| Glucocorticoidresistant isolates | ||||
| 128 | 3.7 | 11 | 73.6 | |
| 129 | 4.3 | 3.3 | 33.3 | |
| 130 | 4.1 | 10.8 | 89 | |
| 136 | 4.8 | 16.8 | 89.4 | |
| 138 | 19.3 | 16.3 | 59.8 | |
| 141 | 5.2 | 13 | 96.9 | |
| 143 | 0.9 | 0.7 | 36.3 | |
| 164 | 0.7 | 7 | 74.7 | |
| 165 | 0.7 | 1.8 | 64.2 | |
| 187 | 1.8 | 4.3 | 55 | |
| 215 | 12.8 | 22.2 | 90.6 | |
| 223 | 6.8 | 9.9 | 87.7 | |
| 240 | 8 | 6.9 | 83 | |
| 241 | 3.2 | 19.4 | 94.3 | |
| 242 | 1.3 | 3.1 | 82 | |
| 243 | 0.9 | 7.8 | 75.2 | |
| 244 | 1.9 | 15.9 | 94.5 | |
| 245 | 3.1 | 8.6 | 45.6 | |
| 248 | 2.3 | 3.4 | 75 | |
| 249 | 0.9 | 3.7 | 67.8 | |
| 253 | 7.2 | 17.3 | 90.5 | |
| Mean (SD) | 4 (4) | 10 (6) | 74 (19) |
| . | Patient Number . | Control* . | MPS . | MG132 . |
|---|---|---|---|---|
| Spontaneously apoptotic isolates | ||||
| 156 | 46.1 | 60.5 | 87.3 | |
| 160 | 27.4 | 34.1 | 82.7 | |
| 180 | 89.8 | 90.1 | 96.1 | |
| 213 | 24.9 | 48.5 | 70.1 | |
| 227 | 37.9 | 49.8 | 93.8 | |
| 228 | 51.2 | 71.2 | 97.3 | |
| 230 | 32.4 | 57.7 | 90.4 | |
| 232 | 25.6 | 37.8 | 91.2 | |
| 233 | 29.5 | 31.8 | 72.8 | |
| 235 | 38.5 | 57.6 | 89.9 | |
| Mean (SD) | 40 (19) | 54 (18) | 87 (9) | |
| Glucocorticoidsensitive isolates | ||||
| 131 | 0.7 | 66.3 | 99.5 | |
| 132 | 3.7 | 51 | 95.4 | |
| 134 | 8.9 | 23.4 | 99.6 | |
| 135 | 15.7 | 90.6 | 88.8 | |
| 137 | 11.2 | 37.9 | 88.6 | |
| 140 | 2.4 | 24.3 | 90.4 | |
| 144 | 4.6 | 34.1 | 50.9 | |
| 146 | 13.4 | 31 | 78.5 | |
| 147 | 1.4 | 19.6 | 92.5 | |
| 148 | 10 | 49.4 | 95.2 | |
| 149 | 1.6 | 30.1 | 99 | |
| 154 | 4.1 | 99.4 | 98.3 | |
| 155 | 16.7 | 36.9 | 99.8 | |
| 157 | 8.7 | 33.3 | 55.3 | |
| 188 | 3.3 | 33.3 | 36.8 | |
| 209 | 8 | 34.8 | 95.7 | |
| 210 | 3.7 | 37.1 | 88.9 | |
| 211 | 8.7 | 48.9 | 98.6 | |
| 212 | 13.8 | 41.8 | 89.3 | |
| 214 | 3.1 | 20.4 | 92.9 | |
| 218 | 2.3 | 23.5 | 96.7 | |
| 220 | 7.6 | 30.3 | 95 | |
| 222 | 2.5 | 30.5 | 80.1 | |
| 229 | 10.7 | 44.9 | 77.2 | |
| 238 | 5.2 | 31.7 | 92.2 | |
| 247 | 3 | 44.8 | 95.7 | |
| 250 | 18.5 | 33.2 | 82.7 | |
| 251 | 16.1 | 61.6 | 82.9 | |
| Mean (SD) | 7 (5) | 41 (19) | 87 (15) | |
| Glucocorticoidresistant isolates | ||||
| 128 | 3.7 | 11 | 73.6 | |
| 129 | 4.3 | 3.3 | 33.3 | |
| 130 | 4.1 | 10.8 | 89 | |
| 136 | 4.8 | 16.8 | 89.4 | |
| 138 | 19.3 | 16.3 | 59.8 | |
| 141 | 5.2 | 13 | 96.9 | |
| 143 | 0.9 | 0.7 | 36.3 | |
| 164 | 0.7 | 7 | 74.7 | |
| 165 | 0.7 | 1.8 | 64.2 | |
| 187 | 1.8 | 4.3 | 55 | |
| 215 | 12.8 | 22.2 | 90.6 | |
| 223 | 6.8 | 9.9 | 87.7 | |
| 240 | 8 | 6.9 | 83 | |
| 241 | 3.2 | 19.4 | 94.3 | |
| 242 | 1.3 | 3.1 | 82 | |
| 243 | 0.9 | 7.8 | 75.2 | |
| 244 | 1.9 | 15.9 | 94.5 | |
| 245 | 3.1 | 8.6 | 45.6 | |
| 248 | 2.3 | 3.4 | 75 | |
| 249 | 0.9 | 3.7 | 67.8 | |
| 253 | 7.2 | 17.3 | 90.5 | |
| Mean (SD) | 4 (4) | 10 (6) | 74 (19) |
Apoptosis was quantified by PI staining and FACS analysis after 16 hours of culture as described in Materials and Methods. Spontaneously apoptotic isolates were arbitrarily defined as those exhibiting greater than 20% DNA fragmentation in control (untreated) cultures. Glucocorticoid-sensitive isolates were arbitrarily defined as those exhibiting greater than 20% increase in DNA fragmentation relative to controls following exposure to steroid.
Abbreviation: MPS, methylprednisolone.
For spontaneously apoptotic isolates, 16-hour control was used.