Table 2.
Effect of Pter Treatment on Different Nrf2- and Redox State-Related Enzyme Activities and Metabolites in Melanoma Cells Growing In Vivo
| A2058 | MeWo | MelJuso | ||||
|---|---|---|---|---|---|---|
| − | + Pter | − | + Pter | − | + Pter | |
| GSH and TXN | ||||||
| GCL (mU/106 cells) | 133 ± 25 | 58 ± 17† | 77 ± 19 | 35 ± 12† | 112 ± 31 | 49 ± 12† |
| GSS (mU/106 cells) | 16.3 ± 4.7 | 8.5 ± 2.0* | 12.4 ± 3.6 | 7.4 ± 1.6* | 13.6 ± 2.4 | 7.8 ± 1.5† |
| GPX (mU/106 cells) | 6.1 ± 2.2 | 3.9 ± 1.1 | 4.6 ± 2.0 | 2.8 ± 1.0 | 5.3 ± 1.7 | 2.1 ± 0.5† |
| GSR (mU/106 cells) | 10.0 ± 3.0 | 6.1 ± 1.6 | 8.5 ± 1.7 | 4.9 ± 1.2* | 9.0 ± 3.0 | 5.1 ± 1.0† |
| GST (mU/106 cells) | 15.1 ± 2.8 | 7.5 ± 1.3† | 10.3 ± 2.2 | 6.5 ± 1.3* | 7.8 ± 1.5 | 4.7 ± 1.3* |
| GGT (mU/106 cells) | 20.5 ± 4.5 | 19.6 ± 2.7 | 15.3 ± 3.7 | 16.0 ± 2.4 | 17.5 ± 3.4 | 15.9 ± 3.5 |
| GSH (nmol/106 cells) | 22.4 ± 3.9 | 10.5 ± 2.1† | 12.3 ± 2.9 | 7.3 ± 1.7† | 19.6 ± 3.7 | 10.8 ± 2.9† |
| GSSG (nmol/106 cells) | 0.3 ± 0.1 | 0.3 ± 0.1 | 0.1 ± 0.05 | 0.1 ± 0.03 | 0.1 ± 0.03 | 0.1 ± 0.04 |
| TXN (μg/106 cells) | 1.0 ± 0.3 | 0.5 ± 0.1* | 1.4 ± 0.4 | 0.8 ± 0.2* | 0.9 ± 0.2 | 0.3 ± 0.1† |
| TXNRD (U/106 cells) | 1.3 ± 0.3 | 0.7 ± 0.2* | 1.6 ± 0.3 | 0.9 ± 0.2† | 1.2 ± 0.3 | 0.6 ± 0.1† |
| ROS | ||||||
| SOD1 (U/106 cells) | 1.5 ± 0.5 | 0.6 ± 0.2† | 1.1 ± 0.3 | 0.5 ± 0.2* | 0.8 ± 0.2 | 0.4 ± 0.2* |
| SOD2 (U/106 cells) | 0.3 ± 0.1 | 0.1 ± 0.05† | 0.2 ± 0.1 | 0.1 ± 0.05 | 0.2 ± 0.05 | 0.1 ± 0.05* |
| CAT (mU/106 cells) | 3.8 ± 0.7 | 1.7 ± 0.3† | 2.7 ± 0.4 | 1.6 ± 0.4† | 3.9 ± 1.1 | 1.1 ± 0.3† |
| NOX (R.L.U./106 cells) | 162 ± 38 | 177 ± 4.5 | 146 ± 37 | 132 ± 25 | 155 ± 26 | 142 ± 37 |
| H2O2 (nmol/106 cells x min) | 1.2 ± 0.4 | 0.7 ± 0.2† | 1.5 ± 0.3 | 0.7 ± 0.2† | 1.6 ± 0.4 | 0.8 ± 0.2† |
| O2• − (ΔFL1, a.u.) | 2.4 ± 0.5 | 3.9 ± 0.6* | 2.9 ± 0.8 | 3.5 ± 0.7 | 3.2 ± 0.7 | 5.0 ± 1.3* |
| NADPH supplying dehydrogenases | ||||||
| G6PDH (mU/106 cells) | 660 ± 113 | 305 ± 78† | 437 ± 55 | 266 ± 39† | 512 ± 49 | 366 ± 63† |
| ME (mU/106 cells) | 76 ± 16 | 43 ± 12† | 44 ± 11 | 32 ± 11 | 38 ± 12 | 25 ± 7† |
| IDH (U/106 cells) | 2.2 ± 0.5 | 1.2 ± 0.4* | 2.4 ± 0.4 | 1.5 ± 0.4* | 2.6 ± 0.4 | 1.5 ± 0.4† |
| Redox state | ||||||
| NADPH (nmol/mg prot) | 0.14 ± 0.2 | 0.06 ± 0.02† | 0.12 ± 0.06 | 0.05 ± 0.002* | 0.11 ± 0.3 | 0.05 ± 0.02† |
| NADP+ (nmol/mg prot) | 0.01 ± 0.005 | 0.05 ± 0.01† | 0.01 ± 0.005 | 0.04 ± 0.01† | 0.005 ± 0.001 | 0.03 ± 0.01 |
| GSH/GSSG | 75 ± 6 | 35 ± 5† | 123 ± 11 | 73 ± 11† | 196 ± 21 | 108 ± 17† |
| NADPH/NADP+ | 14.1 ± 2.1 | 1.2 ± 0.3† | 12.0 ± 3.5 | 1.2 ± 0.2† | 22.0 ± 4.2 | 1.7 ± 0.3† |
Tumor-bearing mice were treated as in Figure 1A. All parameters (see under the Materials and Methods section) were measured in melanoma cells isolated from tumors 35 days after inoculation. Data are mean values ± SD for seven to eight different tumors per parameter and experimental condition.
Significantly different p < 0.05, †p < 0.01.
CAT, catalase; GCL, γ-glutamylcysteine ligase; GGT, γ-glutamyltranspeptidase; GSR, glutathione reductase; GSS, GSH synthase; NOX, NADPH oxidase; GPX, glutathione peroxidase; Nrf2, nuclear factor (erythroid-derived 2)-like 2; ROS, reactive oxygen species; SOD, superoxide dismutase.