Molecular and Cellular Biochemistry

, Volume 368, Issue 1–2, pp 195–201 | Cite as

Up-regulation of interferon-stimulated gene15 and its conjugates by tumor necrosis factor-α via type I interferon-dependent and -independent pathways



Interferon-stimulated gene15 (ISG15) is the first characterized ubiquitin-like protein, which is strongly induced by type I interferons (IFN-α/β), bacterial endotoxin, and cellular stress. Up-regulation of ISG15 is observed in several cancer cell types and is associated with cancer progression. As many cytokines can influence all stages of tumorigenesis, the elevated expression of ISG15 system may be regulated in cancer cells by inflammatory cytokines. In this study, we showed that TNF-α, but not TGF-β and IL-6, up-regulates levels of both ISG15 and its conjugates in human lung carcinoma A549 and human squamous carcinoma HSC4 cell lines. Induction of ISG15 and its conjugates by TNF-α was dose-dependent and required mediation of p38 MAP kinase and Jak1 through up-regulation of endogenous type I interferon expression. SB202190 (p38 MAPK inhibitor) and Jak1 inhibitor suppressed TNF-α-induced expression of ISG15 and its conjugates. However, only SB202190 inhibited the expression of type I interferons by TNF-α. Although B18R, a soluble type I interferon receptor, totally abolished the effect of exogenous IFN-β, it was unable to inhibit completely the TNF-α-induced ISG15 production. In addition, the initiation of ISG15 induction by TNF-α was detected earlier than that of IFN-β induction. Taken together, TNF-α elicits the induction of ISG15 and ISG15 conjugates not only via the autocrine stimulation of type I interferon expression, but also through a type I interferon-independent pathway. These data provide a possible link between inflammatory response and cancer progression.


Cancer Interferon-stimulated gene15 Tumor necrosis factor-α Type I interferon 



This work was supported by grants from the Thailand Research Fund (MRG5180019), Faculty of Dentistry, Mahidol University, and Faculty of Medicine Research Fund, Chiang Mai University, Thailand. The authors would like to thank Professor P. Wilairat, Faculty of Science, Mahidol University for critical reading of the manuscript and Mrs. S. Korsuwannawong for assistance in statistical analysis.

Supplementary material

11010_2012_1360_MOESM1_ESM.tif (564 kb)
Supplementary Fig. Two-dimensional Western blot analysis of TNF-α and IFN-β induced A549 cells. Cells were treated with 15 ng/ml TNF-α or 103 IU/ml IFN-β at 37oC for 24 h. Fifty µg of total proteins was precipitated with 3 volumes of cold acetone. and analyzed in the first dimension by Ettan IPGphor isoelectric focusing system (GE Healthcare, Piscataway, NJ, USA) using premade immobilized pH gradient (IPG) strips with pH range 3-10 (GE Healthcare, Piscataway, NJ, USA). The Strips were further analyzed by SDS-PAGE. ISG15 and ISG15 conjugates cross-reactive spots were detected by immunoblotting as described in legend to Fig. 1. (TIFF 564 kb)


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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  1. 1.Department of Oral Biology, Faculty of DentistryMahidol UniversityBangkokThailand
  2. 2.Department of Biochemistry, Faculty of MedicineChiang Mai UniversityChiang MaiThailand

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