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Induction of interleukin-8 preserves the angiogenic response in HIF-1α–deficient colon cancer cells

Nature Medicine volume 11, pages 992–997 (2005)Cite this article

A Corrigendum to this article was published on 01 February 2006

Abstract

Hypoxia inducible factor-1 (HIF-1) is considered a crucial mediator of the cellular response to hypoxia through its regulation of genes that control angiogenesis1,2,3,4. It represents an attractive therapeutic target5,6 in colon cancer, one of the few tumor types that shows a clinical response to antiangiogenic therapy7. But it is unclear whether inhibition of HIF-1 alone is sufficient to block tumor angiogenesis8,9. In HIF-1α knockdown DLD-1 colon cancer cells (DLD-1HIF-kd), the hypoxic induction of vascular endothelial growth factor (VEGF) was only partially blocked. Xenografts remained highly vascularized with microvessel densities identical to DLD-1 tumors that had wild-type HIF-1α (DLD-1HIF-wt). In addition to the preserved expression of VEGF, the proangiogenic cytokine interleukin (IL)-8 was induced by hypoxia in DLD-1HIF-kd but not DLD-1HIF-wt cells. This induction was mediated by the production of hydrogen peroxide and subsequent activation of NF-κB. Furthermore, the KRAS oncogene, which is commonly mutated in colon cancer, enhanced the hypoxic induction of IL-8. A neutralizing antibody to IL-8 substantially inhibited angiogenesis and tumor growth in DLD-1HIF-kd but not DLD-1HIF-wt xenografts, verifying the functional significance of this IL-8 response. Thus, compensatory pathways can be activated to preserve the tumor angiogenic response, and strategies that inhibit HIF-1α may be most effective when IL-8 is simultaneously targeted.

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Figure 1: Growth of DLD-1HIF-kd cells in vivo.
Figure 2: Knockdown of HIF-1 facilitates the induction of IL-8 by NF-κB during hypoxic conditions.
Figure 3: Increased production of ROS in HIF-knockdown cells expressing KRAS.
Figure 4: Role of IL-8 in tumor angiogenesis in vivo.

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Acknowledgements

We thank the following individuals for sharing these plasmids: C. Reinecker (IL-8 reporter construct), R. Xavier (NF-κB reporter construct, phr-GFP-K-rasVal12) and D. Tenen (pRL-null). We also thank Y. Kamegaya, M. Takeda, M. Ii, E. di Tomaso, T. Padera, P. Au and R. Tyszkowski for assistance with tissue analysis. DNA microarray studies were performed at the DNA Microarray Core Facility at the Massachusetts General Hospital Cancer Center. Confocal microscopy was performed through the Imaging Core of the Center for Study of Inflammatory Bowel Diseases. This work was supported by US National Institutes of Health (NIH) research grant CA92594 to D.C.C. O.I. was supported by NIH grant CA104574, B.R.R. was supported by NIH grant CA098333, M.A.Z. was supported by von Hippel-Lindau Family Alliance, M.G. was supported by an American Gastroenterology Association student fellowship award, and E.-M.D. was supported by a postdoctoral fellowship award from the Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Gastrointestinal Unit, Massachusetts General Hospital and Harvard Medical School, 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Yusuke Mizukami, Won-Seok Jo, Eva-Maria Duerr, Manish Gala, Jingnan Li, Xiaobo Zhang & Daniel C Chung

  2. Department of Medicine, Oncology Unit, Massachusetts General Hospital and Harvard Medical School, 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Michael A Zimmer & Othon Iliopoulos

  3. Department of Pathology, Massachusetts General Hospital and Harvard Medical School, 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Lawrence R Zukerberg

  4. Third Department of Internal Medicine, Asahikawa Medical College, 2-1 Midorigaoka-Higashi, Asahikawa, 078-8510, Japan

    Yutaka Kohgo

  5. Department of Obstetrics and Gynecology, Vincent Center for Reproductive Biology, Massachusetts General Hospital and Harvard Medical School, 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Maureen P Lynch & Bo R Rueda

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  1. Yusuke Mizukami
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Correspondence to Daniel C Chung.

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Supplementary information

Supplementary Fig. 1

Growth of Caco2HIF-wt and Caco2HIF-kd cells in virto ( a ) and in in vivo as xenografts in nude mice ( b ). (PDF 774 kb)

Supplementary Fig. 2

Immunoblot of HIF-1α and HIF-2α proteins in DLD-1HIF-WT, DLD-1 HIF-kd/1470 and DLD-1 HIF-kd/2192 cells in normoxic (N) and hypoxic (H) conditions. (PDF 1022 kb)

Supplementary Fig. 3

The hypoxic induction of IL-8 in the absence of HIF-1α is observed in many cancer cell lines. (PDF 1131 kb)

Supplementary Fig. 4

Specificity of the pSR.HIF-1α1470 and pSR.HIF-1α2192 constructs was demonstrated by coexpression of HIF-1α expression vectors with synonymous codon mutations that are not affected by the siRNA target sequences. (PDF 1320 kb)

Supplementary Fig. 5

Oncogenic KRAS plays a role in the hypoxic induction of IL-8 in non-colonic cancers. (PDF 1157 kb)

Supplementary Table 1

Hypoxic gene expression patterns in DLD-1 cells with or without HIF-1. (PDF 68 kb)

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Mizukami, Y., Jo, WS., Duerr, EM. et al. Induction of interleukin-8 preserves the angiogenic response in HIF-1α–deficient colon cancer cells. Nat Med 11, 992–997 (2005). https://doi.org/10.1038/nm1294

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