Table 2.
Transcriptional Regulation of Genes
| Uridine | Redoxal | Redoxal + uridine | Stavudine | Stavudine + uridine | Stavudine + redoxal | Stavudine + redoxal + uridine | |
|---|---|---|---|---|---|---|---|
| Cell cycle | |||||||
| PCNA | 1.2 ± 0.1 | 1.0 ± 0.1 | 0.8 ± 0.2 | 1.0 ± 0.1 | 0.8 ± 0.3 | 1.2 ± 0.1 | 1.1 ± 0.1 |
| UbcH10 | 1.2 ± 0.1* | 1.1 ± 0.1 | 1.0 ± 0.2 | 1.2 ± 0.1* | 0.8 ± 0.3 | 1.6 ± 0.1* | 1.4 ± 0.2* |
| p21 | 1.4 ± 0.1 | 2.7 ± 0.2** | 2.2 ± 0.4* | 2.1 ± 0.7* | 1.9 ± 0.1* | 4.2 ± 1.4* | 2.7 ± 0.2*** |
| CDKN2A | 1.2 ± 0.2 | 2.1 ± 0.6* | 1.3 ± 0.7 | 3.6 ± 0.9** | 1.9 ± 0.4*††† | 8.1 ± 3.3** | 2.2 ± 0.7*† |
| Apoptosis | |||||||
| BAD | 1.4 ± 0.1 | 2.7 ± 0.6* | 1.7 ± 0.4*† | 2.1 ± 1.0* | 1.8 ± 0.1* | 4.3 ± 1.8* | 2.7 ± 0.1*** |
| BCL2 | 1.5 ± 0.1 | 3.2 ± 0.1*** | 1.7 ± 0.3*††† | 2.0 ± 0.5* | 1.9 ± 0.2* | 4.3 ± 1.3* | 2.6 ± 0.2*** |
| BCL2L1 | 1.1 ± 0.1 | 2.7 ± 0.1*** | 0.9 ± 0.2 | 1.4 ± 0.1* | 1.1 ± 0.1† | 3.6 ± 0.1*** | 1.4 ± 0.3††† |
| BAX | 1.1 ± 0.1 | 1.4 ± 0.1* | 0.9 ± 0.2† | 1.1 ± 0.1 | 1.0 ± 0.1 | 1.9 ± 0.2* | 1.1 ± 0.1†† |
| FAS | 1.6 ± 0.4 | 4.9 ± 1.4* | 1.0 ± 0.1††† | 1.2 ± 0.2 | 1.4 ± 0.3 | 6.6 ± 0.3*** | 1.6 ± 0.1**††† |
| FASLG | 1.5 ± 0.2 | 3.5 ± 0.4** | 2.5 ± 0.7* | 2.5 ± 1.2 | 1.8 ± 0.1*† | 7.0 ± 2.1** | 3.8 ± 0.9** |
| CASP3 | 1.1 ± 0.1 | 2.0 ± 0.1** | 1.0 ± 0.2†† | 1.2 ± 0.1 | 1.1 ± 0.1 | 3.0 ± 0.1*** | 1.4 ± 0.1**††† |
| CASP8 | 1.2 ± 0.1 | 1.9 ± 0.1* | 1.0 ± 0.1†† | 1.4 ± 0.1* | 1.1 ± 0.1† | 2.5 ± 0.2** | 1.3 ± 0.2†† |
| CASP9 | 1.3 ± 0.1* | 2.2 ± 0.1*** | 0.8 ± 0.1††† | 1.3 ± 0.1* | 1.3 ± 0.1* | 3.6 ± 0.4** | 1.7 ± 0.2**†† |
| Mitochondrial energy metabolism | |||||||
| COX IV | 1.2 ± 0.1 | 2.1 ± 0.1** | 1.0 ± 0.2† | 1.2 ± 0.1 | 1.2 ± 0.1* | 2.8 ± 0.2** | 1.5 ± 0.2†† |
| ATP9 | 1.3 ± 0.1 | 3.1 ± 0.3** | 0.8 ± 0.4† | 1.4 ± 0.1* | 1.0 ± 0.2† | 4.3 ± 0.1*** | 1.7 ± 0.2**††† |
| 16S rRNA | 1.2 ± 0.4 | 2.5 ± 1.1* | 0.6 ± 0.1† | 4.1 ± 1.3** | 1.3 ± 0.7††† | 2.6 ± 0.6** | 1.2 ± 0.8* |
| ND1 | 0.8 ± 0.2 | 1.3 ± 0.2* | 0.9 ± 0.1 | 1.1 ± 0.3 | 0.6 ± 0.1* | 2.0 ± 0.6* | 1.0 ± 0.4* |
| COX I | 0.9 ± 0.1 | 1.3 ± 0.4 | 0.6 ± 0.3 | 0.8 ± 0.3 | 1.1 ± 0.2 | 2.3 ± 0.4* | 1.0 ± 0.2†† |
| ATP6 | 0.9 ± 0.1 | 1.6 ± 0.4 | 1.1 ± 0.2† | 0.8 ± 0.1 | 1.2 ± 0.6 | 2.9 ± 0.6** | 0.7 ± 0.1* |
| ND6 | 1.0 ± 0.1 | 2.7 ± 1.1* | 1.5 ± 0.2*† | 1.1 ± 0.1 | 1.6 ± 0.5 | 5.3 ± 2.4* | 1.2 ± 0.8*† |
| Mitochondrial biogenesis | |||||||
| POLG | 1.0 ± 0.1 | 1.7 ± 0.5 | 0.9 ± 0.1† | 1.1 ± 0.2 | 1.4 ± 0.5 | 2.3 ± 0.8* | 1.7 ± 0.8* |
| PGC-1 | 1.6 ± 0.1 | 3.0 ± 0.4** | 2.2 ± 0.3* | 2.4 ± 0.1* | 2.4 ± 0.1* | 5.3 ± 2.3* | 3.8 ± 0.3*** |
| TFAM | 1.7 ± 0.6 | 1.9 ± 0.6 | 1.2 ± 0.4† | 1.2 ± 0.2 | 1.2 ± 0.2 | 1.8 ± 0.6 | 1.4 ± 0.3 |
| Nucleotide metabolism | |||||||
| DHFR | 0.9 ± 0.1 | 1.9 ± 0.4* | 0.5 ± 0.2*† | 0.9 ± 0.1 | 0.9 ± 0.1 | 2.6 ± 0.6* | 1.0 ± 0.3*† |
| DGUOK | 1.1 ± 0.1* | 1.9 ± 0.1** | 0.9 ± 0.1†† | 1.1 ± 0.1 | 1.0 ± 0.1 | 2.6 ± 0.3** | 1.2 ± 0.2†† |
| HPRT1 | 1.1 ± 0.1 | 1.1 ± 0.1 | 0.9 ± 0.2 | 1.2 ± 0.1* | 1.1 ± 0.1† | 1.6 ± 0.1** | 1.3 ± 0.2* |
| DHODH | 1.0 ± 0.2 | 0.8 ± 0.1 | 0.9 ± 0.2† | 1.1 ± 0.1 | 1.0 ± 0.1 | 1.3 ± 0.1** | 1.2 ± 0.1 |
| UMPS | 1.2 ± 0.1* | 1.6 ± 0.1* | 1.0 ± 0.2†† | 1.1 ± 0.1 | 0.8 ± 0.2† | 2.4 ± 0.1* | 1.3 ± 0.1*††† |
| RRM1 | 1.0 ± 0.1* | 1.6 ± 0.3* | 0.8 ± 0.2† | 1.1 ± 0.1 | 1.0 ± 0.1† | 2.2 ± 0.2* | 1.1 ± 0.1*††† |
| TP | 1.5 ± 0.1* | 2.5 ± 0.1*** | 1.7 ± 0.4*† | 4.8 ± 2.2* | 1.8 ± 0.1*††† | 11.0 ± 3.4* | 2.5 ± 0.1***† |
| UPP1 | 1.1 ± 0.1* | 1.4 ± 0.4 | 0.7 ± 0.3† | 1.4 ± 0.1* | 0.8 ± 0.2† | 2.4 ± 0.9* | 1.4 ± 0.2† |
| UCK2 | 1.2 ± 0.1 | 1.3 ± 0.1* | 1.0 ± 0.2 | 1.2 ± 0.1 | 1.1 ± 0.1 | 1.9 ± 0.1** | 1.4 ± 0.2 |
| TK1 | 0.9 ± 0.3* | 1.6 ± 0.1* | 0.9 ± 0.2† | 1.2 ± 0.1 | 1.1 ± 0.1† | 2.3 ± 0.1** | 1.5 ± 0.2*† |
| DCK | 1.0 ± 0.1 | 1.6 ± 0.1* | 0.6 ± 0.2† | 0.9 ± 0.2 | 1.0 ± 0.1 | 2.6 ± 0.7* | 1.2 ± 0.3† |
| Oxidative stress | |||||||
| GPX1 | 2.7 ± 2.4 | 5.2 ± 3.5 | 3.6 ± 1.7 | 4.6 ± 2.7 | 4.4 ± 3.1 | 8.3 ± 6.1 | 5.0 ± 3.8 |
| SOD1 | 1.1 ± 0.1 | 2.1 ± 0.1** | 0.9 ± 0.1††† | 1.4 ± 0.1* | 0.8 ± 0.3† | 2.1 ± 0.5* | 1.1 ± 0.4* |
| SOD2 | 0.9 ± 0.4 | 2.1 ± 0.3* | 1.4 ± 0.2† | 1.1 ± 0.2 | 1.8 ± 0.2*† | 2.6 ± 0.1*** | 1.9 ± 1.1* |
| DNA repair | |||||||
| UNG1 | 1.1 ± 0.1* | 0.9 ± 0.1 | 1.0 ± 0.2 | 1.0 ± 0.1 | 1.1 ± 0.1 | 2.0 ± 0.2* | 1.3 ± 0.2*† |
| OGG1 | 1.2 ± 0.1* | 1.6 ± 0.2* | 1.0 ± 0.2† | 1.3 ± 0.1* | 1.3 ± 0.1* | 3.1 ± 0.3** | 1.8 ± 0.2**† |
| APE1 | 1.1 ± 0.1 | 1.8 ± 0.1* | 0.9 ± 0.2†† | 1.2 ± 0.1* | 1.1 ± 0.1 | 2.4 ± 0.2** | 1.3 ± 0.2†† |
Values represent the fold difference (±SD) of mRNA copies after 10 days of HepG2 treatment with agents by comparison with unexposed cells. Three independent cell cultures were measured per agent added. Significance levels:
P < 0.05,
P < 0.01,
P < 0.001 in comparison to controls (no uridine and no stavudine),
P < 0.05,
P < 0.01,
P < 0.001 in comparison to treatment without uridine. Bold entries represent significant up-regulation by a minimum factor of 2.
Gene abbreviations: LMNB1, lamin B1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ACTB, actin beta; 36B4, ribosomal phosphoprotein P0; PCNA, proliferating cell nuclear antigen; UbcH10, ubiquitin-conjugating enzyme H10; p21, cyclin-dependent kinase inhibitor 1A; CDKN2A, cyclin-dependent kinase inhibitor 2A; BAD, BCL2 antagonist of cell death; BCL2, B-cell lymphoma protein 2; BCL2L1, BCL antagonist of cell death; BAX, BCL2-associated X protein; FAS, fast apoptotic stimulus; FASLG, Fas ligand; CASP3, caspase 3; CASP8, caspase 8; CASP9, caspase 9; COX IV, cytochrome c oxidase subunit IV; ATP9, ATP synthase subunit 9; 16S rRNA; ND1, NADH dehydrogenase subunit 1, complex I; COX I, cytochrome c-oxidase subunit 1; ATP6, ATP synthase subunit 6; ND6, NADH dehydrogenase subunit 6; POLG, polymerase-γ (catalytic subunit); PGC-1, peroxisome proliferator-activated γ co-activator-1; TFAM, mitochondrial transcription factor A; DHFR, dihydrofolate reductase; DGUOK, deoxyguanosine kinase; HPRT1, hypoxanthine phosphoribosyltransferase 1; DHODH, dihydroorotate dehydrogenase; UMPS, uridine monophosphate synthetase; RRM1, ribonucleotide reductase M1; TP, thymidine phosphorylase; UPP1, uridine phosphorylase 1; UCK2, uridine-cytidine kinase 2; TK1, thymidine kinase 1; DCK, deoxycytidine kinase; GPX1, glutathione peroxidase 1; SOD1, superoxide dismutase 1; SOD2, superoxide dismutase 2; UNG1, uracil-DNA glycosylase; OGG1, 8-oxoguanine DNA glycosylase; APE1, apurinic/apyrimidinic endonuclease 1.