Table 1.
Effect of 24 h incubation with 5 mM ThDP on the assayed metabolic parameters of A549wt and A549p21- cells in the standard or thiamine-free DMEM. The data of at least three independent experiments are presented as means ±SEM. The eight experimental groups are marked by the letters (a–h) in the “Cell line” row. Statistically significant differences (p < 0.05) are determined by two-way ANOVA followed by Tukey’s post hoc test and are indicated as the difference of considered parameter to that in the corresponding experimental groups (a–h). Because the reaction rates of cellular NAD(P)H:XTT reductase are relative to the corresponding control cells, the comparison of this parameter is done only for different conditions applied to either A549wt or A549p21- cells. GSH—glutathione, GSSG—glutathione disulfide.
| Medium | Standard DMEM | Thiamine-free DMEM | ||||||
|---|---|---|---|---|---|---|---|---|
| Addition of ThDP for 24h | - | + 5 mM ThDP | - | + 5 mM ThDP | ||||
| Cell Line | A549wt a | A549p21- b | A549wt c | A549p21- d | A549wt e | A549p21- f | A549wt g | A549p21- h |
| Protein, mg | 1.27 ± 0.03 | 1.14 ± 0.07 e,h | 1.08 ± 0.05 e,f,h | 1.13 ± 0.02 e,h | 1.46 ± 0.04 b,c,d,g | 1.38 ± 0.07 c | 1.17 ± 0.05 e,h | 1.44 ± 0.07 b,c,d,g |
| Activity, μmol/(min∙mg) | ||||||||
| total OGDHC∙104 | 3.3 ± 0.8 e,g,h | 5.5 ± 1.4 e,f,g | 4.3 ± 0.3 e,f,g,h | 5.9 ± 0.4 e,f,g | 1.5 ± 0.7 a,b,c,d,g,h | 1.5 ± 0.2 b,c,d,g,h | 8.7 ± 0.2 a,b,c,d,e,f | 8.3 ± 0.3 a,c,e,f |
| apo-OGDHC∙104 | 2.5 ± 0.8 c,e | 3.7 ± 1.4 c,e,f | 0.2 ± 0.3 a,b,g | 2.6 ± 0.4 e,f | 0 ± 0.2 a,b,d,g,h | 0.1 ± 0.2 b,d,g,h | 3.4 ± 0.2 c,e,f | 2.3 ± 0.3 e,f |
| holo-OGDHC∙104 | 0.7 ± 0.3 c,d,g,h | 1.8 ± 0.6 c,g,h | 4.1 ± 0.4 a,b,f | 3.3 ± 0.5 a,h | 2.0 ± 0.3 g,h | 1.4 ± 0.4 c,g,h | 5.3 ± 0.5 a,b,e,f | 6.0 ± 0.5 a,b,d,e,f |
| GSH, nmol/mg | 38 ± 3 e,f | 41 ± 1 f | 29 ± 2 e,f | 28 ± 3 e,f,h | 55 ± 5 a,c,d | 61 ± 5 a,b,c,d,g,h | 43 ± 2 f | 43 ± 2 d,f |
| GSSG, nmol/mg | 1.9 ± 0.1 e,f,g,h | 2.6 ± 0.2 f | 2.2 ± 0.1 f | 2.1 ± 0.1 f,h | 2.8 ± 0.2 a | 3.4 ± 0.1 a,b,c,d | 2.7 ± 0.1 a | 2.9 ± 0.3 a,d |
| GSH/GSSG | 19.4 ± 0.1 c,d | 15.7 ± 0.8 | 13.3 ± 1.4 a,e | 13.8 ± 1.6 a,e | 20.2 ± 0.5 c,d | 17.9 ± 1.5 | 15.6 ± 0.4 | 15.2 ± 1.4 |
| p53, % | 100 ± 10 d,g,h | 110 ± 22 d,g,h | 129 ± 9 | 210 ± 52 a,b,f | 166 ± 8 f | 72 ± 8 d,e,g,h | 209 ± 22 a,b,f | 212 ± 4 a,b,f |
| NAD(P)H:XTT reductase, % | 100 ± 2 c,e,g | 100 ± 3 d,f,h | 33 ± 2 a,g | 40 ± 3 b | 46 ± 1 a | 62 ± 1 b | 65 ± 1 a,c | 47 ± 1 b |