Involvement of the IFNγ-JAK1-STAT1 pathway in the transcriptional response to doxorubicin
HeLa cells were pretreated for 1.5 h with or without neutralizing antibodies against IFNγ or INFγ receptor 1, ATA (30 μM), or AG490 (50 μM) followed by treatment with 1 μM doxorubicin for 48 h. RNA was extracted, reverse-transcribed, and analyzed by qRT-PCR. Gene expression is calculated as the ratio of 18S normalized fluorescence intensities measured in the respective DMSO controls. Gene expression levels upon doxorubicin exposure was set to 100%, and assays containing inhibitors were normalized to this value. Mean ± S.D., n = 3.
| Percentage of Dox | Dox + Anti-IFNγ-Ab | Dox + Anti-IFNγR1-Ab | Dox + 30 μM ATA | Dox + 50 μM AG490 |
|---|---|---|---|---|
| STAT1 | 48.3 ± 3.0*** | 83.4 ± 23.5 | 178.5 ± 32.8 | 41.1 ± 4.8* |
| STAT2 | 86.2 ± 39.1 | 88.5 ± 43.9 | 61.9 ± 4.6** | 53.0 ± 16.0* |
| IFI6-16 | 43.3 ± 16.8* | 62.7 ± 14.4* | 31.7 ± 2.7* | 29.5 ± 7.8* |
| IFI30 | 43.9 ± 20.8* | 55.5 ± 28.1 | 42.1 ± 17.9* | 27.8 ± 7.3* |
| IFI35 | 34.6 ± 2.8* | 56.2 ± 13.6* | 22.0 ± 2.1** | 48.0 ± 13.2* |
| IFITM1 | 23.0 ± 7.1* | 32.7 ± 6.3* | 37.0 ± 8.8* | 48.7 ± 9.6** |
| ISGF3G/IRF9 | 36.3 ± 10.2** | 39.4 ± 17.2* | 63.4 ± 30.5* | 195 ± 45.7 |
| NMI | 35.5 ± 12.6** | 46.9 ± 9.2* | 41.5 ± 5.6* | 39.2 ± 5.9* |
| Caspase 1 | 24.2 ± 12.4* | 45.6 ± 5.2* | 32.6 ± 8.3** | 74.2 ± 5.0* |