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Ras regulates interleukin-1β-induced HIF-1α transcriptional activity in glioblastoma

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Abstract

We observed elevated levels of pro-inflammatory cytokine IL-1β in glioblastoma multiforme tumor samples. Since hypoxia-inducible factor-1α (HIF-1α) plays a crucial role in linking inflammatory and oncogenic pathways, we investigated the effect of IL-1β on HIF-1α expression in glioma cells under normoxia. IL-1β-mediated elevation of HIF-1α transcriptional activity was dependent on Ras-induced NF-κB activation, as IL-1β failed to induce NF-κB and HIF-1α activity in cells transfected with dominant negative RasN17. Increased Ras expression was accompanied by increased phosphorylation of Ras effectors AKT, ERK, JNK, and p38MAPK. While inhibition of these effectors individually failed to block the IL-1β-mediated increase in HIF-1α induction, co-inhibition of both AKT and ERK resulted in a significant decrease in IL-1β-induced HIF-1α activation. Interestingly, IL-1β elevated Wnt-1 expression in a Ras-dependent manner, and small interfering RNA (siRNA)-mediated knockdown of Wnt-1 decreased HIF-1α activity. Although Wnt-1-mediated HIF-1α was independent of the canonical Wnt/β-catenin signaling pathway, it regulated HIF-1α through NF-κB. siRNA-mediated HIF-1α knockdown attenuated elevated IL-1β mRNA levels induced upon IL-1β treatment. This was accompanied by increased interaction of HIF-1α with HIF responsive element on the IL-1β promoter upon IL-1β treatment, under normoxia. Our studies highlights for first time that (1) Ras is a key mediator of IL-1β-induced NF-κB and HIF-1α activation, under normoxia; (2) Wnt-1 regulates IL-1β-mediated HIF-1α induction via NF-κB; (3) Ras and Wnt-1 are intermediaries in the canonical IL-1β–NF-κB signaling pathway downstream of MyD88; and (4) IL-1β-induced HIF-1α drives a HIF-1α-IL-1β autocrine loop to maintain persistently elevated IL-1β level.

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Acknowledgements

The work was supported by a research grant from the Department of Biotechnology (BT/PR7047/Med/14/934/2006) and partly by a research grant by the Defence Research and Development Organization DRDO (DLS/81/148222LSRB-140/EPB/2007), Government of India, to ES. VS, DD, and NK are supported by a research fellowship from Council of Scientific and Industrial Research (CSIR, Government of India). We would like to thank Uttam Kumar Saini for technical assistance and Mr. R Khader Valli for help with Real-Time PCR.

Conflict of interest

There are no competing financial interests in relation to the work and we have nothing to disclose.

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Authors and Affiliations

  1. National Brain Research Centre, Manesar, Haryana, 122 050, India

    Vivek Sharma, Deobrat Dixit, Nitin Koul & Ellora Sen

  2. Paras Hospitals, Gurgaon, India

    Veer Singh Mehta

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  1. Vivek Sharma
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Correspondence to Ellora Sen.

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Sharma, V., Dixit, D., Koul, N. et al. Ras regulates interleukin-1β-induced HIF-1α transcriptional activity in glioblastoma. J Mol Med 89, 123–136 (2011). https://doi.org/10.1007/s00109-010-0683-5

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