Sensitivity to TNF/DOX and Expression of Intracellular Resistance Factors in Leukemic Cells Obtained From Patients
| . | FAB Type . | Sensitivity (% Cytotoxicity) to* . | MnSOD Activity† . | enTNF Expression‡ . | |
|---|---|---|---|---|---|
| . | . | TNF . | DOX . | (U/mg protein) . | (relative F.I.) . |
| . | . | (100 U/mL) . | (5 μmol/L) . | . | . |
| 1 | ALL (L1) | 60.8 | 52.5 | NT | 1.2 |
| 2 | ALL (L1) | 21.5 | 36.9 | 11.67 ± 0.28 | 1.8 |
| 3 | ALL (L2) | 10.1 | 37.2 | 9.68 ± 2.54 | 2.6 |
| 4 | ALL (L2) | 61.5 | 40.3 | 7.24 ± 0.92 | 0.9 |
| 4′ | ALL (L2) | 12.5 | 18.4 | 11.80 ± 2.21 | 1.9 |
| 5 | ALL (L2) | 51.0 | 53.2 | 7.49 ± 0.54 | 0.6 |
| 6 | ALL (L2) | 50.8 | 58.7 | 9.17 ± 0.42 | 0.9 |
| 7 | AML (M1) | 0.1 | 26.9 | 12.01 ± 2.23 | 1.6 |
| 8 | AML (M1) | 19.3 | 28.5 | 10.66 ± 0.50 | 1.8 |
| 9 | AML (M1) | 1.3 | 1.7 | 8.07 ± 0.64 | 2.0 |
| 10 | AML (M1) | 7.4 | 29.1 | 10.56 ± 0.20 | 2.9 |
| 11 | AML (M1) | 25.1 | 42.9 | 9.15 ± 0.10 | 1.9 |
| 12 | AML (M2) | 3.8 | 24.2 | 11.36 ± 1.85 | 3.1 |
| 13 | AML (M2) | 65.2 | 61.7 | 4.48 ± 0.47 | 0.8 |
| 14 | AML (M2) | 27.5 | 33.4 | 7.63 ± 1.58 | NT |
| 15 | AML (M2) | 13.8 | 11.9 | 10.65 ± 0.16 | 1.3 |
| 16 | AML (M2) | 39.1 | 60.1 | 8.37 ± 0.73 | 0.5 |
| 17 | AML (M2) | 33.9 | 53.9 | 10.08 ± 0.58 | 0.9 |
| 18 | AML (M3) | 28.5 | 32.3 | 7.66 ± 1.27 | 2.1 |
| 19 | AML (M3) | 2.3 | 10.0 | 12.93 ± 1.23 | 3.7 |
| . | FAB Type . | Sensitivity (% Cytotoxicity) to* . | MnSOD Activity† . | enTNF Expression‡ . | |
|---|---|---|---|---|---|
| . | . | TNF . | DOX . | (U/mg protein) . | (relative F.I.) . |
| . | . | (100 U/mL) . | (5 μmol/L) . | . | . |
| 1 | ALL (L1) | 60.8 | 52.5 | NT | 1.2 |
| 2 | ALL (L1) | 21.5 | 36.9 | 11.67 ± 0.28 | 1.8 |
| 3 | ALL (L2) | 10.1 | 37.2 | 9.68 ± 2.54 | 2.6 |
| 4 | ALL (L2) | 61.5 | 40.3 | 7.24 ± 0.92 | 0.9 |
| 4′ | ALL (L2) | 12.5 | 18.4 | 11.80 ± 2.21 | 1.9 |
| 5 | ALL (L2) | 51.0 | 53.2 | 7.49 ± 0.54 | 0.6 |
| 6 | ALL (L2) | 50.8 | 58.7 | 9.17 ± 0.42 | 0.9 |
| 7 | AML (M1) | 0.1 | 26.9 | 12.01 ± 2.23 | 1.6 |
| 8 | AML (M1) | 19.3 | 28.5 | 10.66 ± 0.50 | 1.8 |
| 9 | AML (M1) | 1.3 | 1.7 | 8.07 ± 0.64 | 2.0 |
| 10 | AML (M1) | 7.4 | 29.1 | 10.56 ± 0.20 | 2.9 |
| 11 | AML (M1) | 25.1 | 42.9 | 9.15 ± 0.10 | 1.9 |
| 12 | AML (M2) | 3.8 | 24.2 | 11.36 ± 1.85 | 3.1 |
| 13 | AML (M2) | 65.2 | 61.7 | 4.48 ± 0.47 | 0.8 |
| 14 | AML (M2) | 27.5 | 33.4 | 7.63 ± 1.58 | NT |
| 15 | AML (M2) | 13.8 | 11.9 | 10.65 ± 0.16 | 1.3 |
| 16 | AML (M2) | 39.1 | 60.1 | 8.37 ± 0.73 | 0.5 |
| 17 | AML (M2) | 33.9 | 53.9 | 10.08 ± 0.58 | 0.9 |
| 18 | AML (M3) | 28.5 | 32.3 | 7.66 ± 1.27 | 2.1 |
| 19 | AML (M3) | 2.3 | 10.0 | 12.93 ± 1.23 | 3.7 |
Abbreviation: NT, not tested.
Cells (5 × 104/50 μL) were cultured with DOX (5 μmol/L) or TNF (100 U/mL) for 96 hours and cytotoxic activity was determined by the MTT assay.
Cells (2 × 106/mL) in 0.05 mmol/L potassium phosphate buffer, pH 7.8, were sonicated for 1 minute. The resulting solution was centrifuged at 3,000 rpm for 5 minutes to obtain cell lysates. MnSOD activities and protein concentrations of the cell lysates were determined by the nitroblue tetrazolium method and the Bio-Rad protein assay. Data presented are the mean ± SD of three separate experiments.
enTNF expression was determined by immunofluorescence assay as described in Materials and Methods. Values represent the normalized enTNF expression of leukemic cells compared with that of KG-1 cells.