Sensitivity to TNF and DOX in pTNFΔpro Transfected Clones
| Strain . | % Cytotoxicity to4-150 . | enTNF Expression4-151 . | MnSOD Activity‡ . | |
|---|---|---|---|---|
| . | TNF . | DOX . | (relative F.I.) . | (U/mg protein) . |
| . | (100 U/mL) . | (5 μmol/L) . | . | . |
| KG-1 | 36.4 | 53.6 | 1.00 | 10.90 |
| KG-1(pSV2neo) KP2 | 32.9 | 53.8 | 1.16 | 10.30 |
| KG-1(pTNFΔpro) KD3 | 10.0 | 27.3 | 5.50 | 24.50 |
| KG-1(pTNFΔpro) KD5 | 9.8 | 26.4 | 6.08 | 25.12 |
| KG-1(pTNFΔpro) KD10 | 10.2 | 28.1 | 4.38 | 21.40 |
| Strain . | % Cytotoxicity to4-150 . | enTNF Expression4-151 . | MnSOD Activity‡ . | |
|---|---|---|---|---|
| . | TNF . | DOX . | (relative F.I.) . | (U/mg protein) . |
| . | (100 U/mL) . | (5 μmol/L) . | . | . |
| KG-1 | 36.4 | 53.6 | 1.00 | 10.90 |
| KG-1(pSV2neo) KP2 | 32.9 | 53.8 | 1.16 | 10.30 |
| KG-1(pTNFΔpro) KD3 | 10.0 | 27.3 | 5.50 | 24.50 |
| KG-1(pTNFΔpro) KD5 | 9.8 | 26.4 | 6.08 | 25.12 |
| KG-1(pTNFΔpro) KD10 | 10.2 | 28.1 | 4.38 | 21.40 |
Cells (5 × 104/50 μL) were cultured with TNF (100 U/mL) and DOX (5 μmol/L) for 96 hours and cytotoxic activity was determined by the MTT assay.
enTNF expression was determined by immunofluorescence assay as described in Materials and Methods. Values are normalized enTNF expression of leukemic cells compared with that of KG-1 cells.
Cells (2 × 106/mL) in 0.05 mol/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 Bio-Rad protein assay.