PRIMA-1Met induced cell death in primary cells
| Sample . | Patient’s characteristics . | Cell death . | Ratio of DR5 expression . | DR5 fold increase . | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Disease . | Status . | Origin . | del17p . | PRIMA1Me . | PRIMA1Met+NAC . | PRIMA1Met+GSH . | PRIMA1Met+BSO . | Nutlin3a . | Control . | PRIMA1Met . | Nutlin3a . | PRIMA1Met . | Nutlin3a . | |
| 1 | pPCL | D | PB | 0% | 80% | — | — | — | 0% | 0.95 | 1.31 | 2.42 | 1.38 | 2.55 |
| 2 | MM | R | BM | 0% | 71% | — | — | — | 0% | 1.46 | 1.51 | 1.83 | 1.03 | 1.25 |
| 3 | sPCL | R | PB | 0% | 29% | 5% | — | — | 19% | 1.31 | 1.32 | 1.62 | 1.01 | 1.24 |
| 4 | sPCL | R | PB | 0% | 13% | 0% | 0% | 92% | 2% | 1.51 | 1.54 | 1.89 | 1.02 | 1.25 |
| 5 | MM | R | BM | 0% | 100% | 0% | 0% | 100% | 0% | 3.07 | 1.30 | 9.92 | 0.42 | 3.23 |
| 6 | pPCL | D | BM | 0% | 14% | 0% | 0% | 74% | 0% | — | — | — | — | — |
| 6’ | pPCL | D | PB | 0% | 6% | 0% | 0% | 98% | 0% | — | — | — | — | — |
| 7 | sPCL | R | PB | 0% | 5% | 0% | 0% | 85% | 50% | — | — | — | — | — |
| 8 | MM | D | BM | 0% | 79% | — | — | — | 0% | 1.12 | 0.98 | 1.54 | 0.87 | 1.38 |
| 9 | MM | D | BM | 0% | 46% | — | — | — | 0% | 1.34 | 1.23 | 1.92 | 0.95 | 1.43 |
| 10 | MM | D | BM | 4% | 100% | 0% | 0% | — | — | 1.26 | 1.09 | — | 0.86 | |
| 11 | MM | D | BM | 4% | 15% | 0% | — | — | — | — | — | — | — | — |
| 12 | MM | D | BM | 5% | 82% | — | — | — | 8% | 1.66 | 1.21 | 8.30 | 0.73 | 5.00 |
| 13 | pPCL | D | BM | 6% | 25% | — | — | — | 20% | 1.04 | 1.20 | 1.34 | 1.15 | 1.29 |
| 14 | MM | D | BM | 6% | 10% | 0% | 0% | — | — | 4.27 | 3.13 | — | 0.73 | |
| 15 | MM | D | BM | 7% | 7% | — | — | — | 13% | 2.73 | 2.54 | 3.78 | 0.93 | 1.38 |
| 16 | MM | R | BM | 39% | 94% | — | — | — | 8% | 3.10 | 1.88 | 3.81 | 0.61 | 1.23 |
| 17 | MM | D | BM | 55% | 98% | — | — | — | — | 2.10 | 1.40 | 2.46 | 0.67 | 1.17 |
| 18 | sPCL | R | PB | 68% | 45% | — | — | — | 27% | 1.12 | 1.11 | 1.10 | 0.99 | 0.98 |
| 19 | pPCL | D | PB | 89% | 87% | — | — | 90% | 19% | 1.23 | 1.13 | 1.40 | 0.92 | 1.14 |
| 20 | MM | D | BM | 90% | 55% | — | — | — | 1% | — | — | — | — | — |
| 21 | sPCL | R | PB | 95% | 97% | — | — | — | 7% | 2.08 | 2.00 | 2.30 | 0.96 | 1.11 |
| 22 | pPCL | D | BM | 96% | 50% | 0% | — | 79% | — | — | — | — | — | — |
| 22’ | pPCL | D | PB | 96% | 51% | 0% | — | 96% | — | — | — | — | — | — |
| 23 | pPCL | D | PB | 97% | 96% | — | — | — | 11% | 1.34 | 0.98 | 1.28 | 0.73 | 0.96 |
| Sample . | Patient’s characteristics . | Cell death . | Ratio of DR5 expression . | DR5 fold increase . | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Disease . | Status . | Origin . | del17p . | PRIMA1Me . | PRIMA1Met+NAC . | PRIMA1Met+GSH . | PRIMA1Met+BSO . | Nutlin3a . | Control . | PRIMA1Met . | Nutlin3a . | PRIMA1Met . | Nutlin3a . | |
| 1 | pPCL | D | PB | 0% | 80% | — | — | — | 0% | 0.95 | 1.31 | 2.42 | 1.38 | 2.55 |
| 2 | MM | R | BM | 0% | 71% | — | — | — | 0% | 1.46 | 1.51 | 1.83 | 1.03 | 1.25 |
| 3 | sPCL | R | PB | 0% | 29% | 5% | — | — | 19% | 1.31 | 1.32 | 1.62 | 1.01 | 1.24 |
| 4 | sPCL | R | PB | 0% | 13% | 0% | 0% | 92% | 2% | 1.51 | 1.54 | 1.89 | 1.02 | 1.25 |
| 5 | MM | R | BM | 0% | 100% | 0% | 0% | 100% | 0% | 3.07 | 1.30 | 9.92 | 0.42 | 3.23 |
| 6 | pPCL | D | BM | 0% | 14% | 0% | 0% | 74% | 0% | — | — | — | — | — |
| 6’ | pPCL | D | PB | 0% | 6% | 0% | 0% | 98% | 0% | — | — | — | — | — |
| 7 | sPCL | R | PB | 0% | 5% | 0% | 0% | 85% | 50% | — | — | — | — | — |
| 8 | MM | D | BM | 0% | 79% | — | — | — | 0% | 1.12 | 0.98 | 1.54 | 0.87 | 1.38 |
| 9 | MM | D | BM | 0% | 46% | — | — | — | 0% | 1.34 | 1.23 | 1.92 | 0.95 | 1.43 |
| 10 | MM | D | BM | 4% | 100% | 0% | 0% | — | — | 1.26 | 1.09 | — | 0.86 | |
| 11 | MM | D | BM | 4% | 15% | 0% | — | — | — | — | — | — | — | — |
| 12 | MM | D | BM | 5% | 82% | — | — | — | 8% | 1.66 | 1.21 | 8.30 | 0.73 | 5.00 |
| 13 | pPCL | D | BM | 6% | 25% | — | — | — | 20% | 1.04 | 1.20 | 1.34 | 1.15 | 1.29 |
| 14 | MM | D | BM | 6% | 10% | 0% | 0% | — | — | 4.27 | 3.13 | — | 0.73 | |
| 15 | MM | D | BM | 7% | 7% | — | — | — | 13% | 2.73 | 2.54 | 3.78 | 0.93 | 1.38 |
| 16 | MM | R | BM | 39% | 94% | — | — | — | 8% | 3.10 | 1.88 | 3.81 | 0.61 | 1.23 |
| 17 | MM | D | BM | 55% | 98% | — | — | — | — | 2.10 | 1.40 | 2.46 | 0.67 | 1.17 |
| 18 | sPCL | R | PB | 68% | 45% | — | — | — | 27% | 1.12 | 1.11 | 1.10 | 0.99 | 0.98 |
| 19 | pPCL | D | PB | 89% | 87% | — | — | 90% | 19% | 1.23 | 1.13 | 1.40 | 0.92 | 1.14 |
| 20 | MM | D | BM | 90% | 55% | — | — | — | 1% | — | — | — | — | — |
| 21 | sPCL | R | PB | 95% | 97% | — | — | — | 7% | 2.08 | 2.00 | 2.30 | 0.96 | 1.11 |
| 22 | pPCL | D | BM | 96% | 50% | 0% | — | 79% | — | — | — | — | — | — |
| 22’ | pPCL | D | PB | 96% | 51% | 0% | — | 96% | — | — | — | — | — | — |
| 23 | pPCL | D | PB | 97% | 96% | — | — | — | 11% | 1.34 | 0.98 | 1.28 | 0.73 | 0.96 |
Primary myeloma cells were purified (samples 2, 5, 6, 8, 9, 11, 12, 14, 15, 16, 17, 20, 22, and 23) or not (samples 1, 3, 4, 7, 10, 13, 18, 19, and 21), with CD138-conjugated magnetic beads, and incubated overnight in RPMI1640 supplemented with 5% fetal calf serum and 3 ng/mL interleukin 6, with or without nutlin3a (10 μM), PRIMA-1Met (10 μM), NAC (5 mM), GSH (5 mM), or BSO (0.5 mM), as indicated. Cell death was assessed by the loss of CD138 staining. Levels of DR5 expression were assessed by direct staining with anti-DR5-PE or control-PE mAb of untreated (control) or PRIMA-1Met- or nutlin3a-treated cells, and was expressed as the ratio of the specific fluorescence mean to that of control staining in untreated and treated cells. Samples 6′ and 22′ were peripheral blood samples from patients 6 and 22 and were not considered for statistical analyses. Dash indicates not done. BM, bone marrow; D, diagnosis; p, primary; PB, peripheral blood; PCL, plasma cell leukemia; R, relapse; s, secondary.