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Hypoxia upregulates osteopontin expression in NIH-3T3 cells via a Ras-activated enhancer

Oncogene volume 24, pages 6555–6563 (2005)Cite this article

Abstract

Osteopontin (OPN) is a secreted phosphoglycoprotein that has been linked to tumor progression and survival in several solid tumors, including head and neck cancers. Previous studies showed that OPN expression is induced by tumor hypoxia, and its plasma levels can serve as a surrogate marker for tumor hypoxia and treatment outcome in head and neck cancer patients. In this study, we investigate the transcriptional mechanism by which hypoxia enhances OPN expression. We found that OPN is induced in head and neck squamous cell carcinoma (HNSCC) cell lines and in NIH3T3 cells by hypoxia at both mRNA and protein levels in a time-dependent manner. Actinomycin D chase experiments showed that hypoxic induction of OPN was not due to increased mRNA stability. Deletion analyses of the mouse OPN promoter regions indicated that a ras-activated enhancer (RAE) located at −731 to −712 relative to the transcription start site was essential for hypoxia-enhanced OPN transcription. Using electrophoretic mobility shift assays with the RAE DNA sequence, we found that hypoxia induced sequence-specific DNA-binding complexes. Furthermore, hypoxia and ras exposure resulted in an additive induction of OPN protein and mRNA levels that appeared to be mediated by the RAE. Induction of OPN through the RAE element by hypoxia is mediated by an Akt-kinase signaled pathway as decreasing Akt levels with dominant negative constructs resulted in inhibition of OPN induction by hypoxia. Taken together, these results have identified a new hypoxia responsive transcriptional enhancer that is regulated by Akt signaling.

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References

  • Ausserer WA, Bourrat-Floeck B, Green CJ, Laderoute KR and Sutherland RM . (1994). Mol. Cell. Biol., 14, 5032–5042.

  • Bedogni B, O’Neill MS, Welford SM, Bouley DM, Giaccia AJ, Denko NC and Powell MB . (2004). Cancer Res., 64, 2552–2560.

  • Brizel DM, Scully SP, Harrelson JM, Layfield LJ, Dodge RK, Charles HC, Samulski TV, Prosnitz LR and Dewhirst MW . (1996). Cancer Res., 56, 5347–5350.

  • Brown JM and Giaccia AJ . (1998). Cancer Res., 58, 1408–1416.

  • Chambers AF, Behrend EI, Wilson SM and Denhardt DT . (1992). Anticancer Res., 12, 43–47.

  • Chan DA, Sutphin PD, Denko NC and Giaccia AJ . (2002). J. Biol. Chem., 277, 40112–40117.

  • Choi H, Lin Z, Li B and Liu A . (1990). J. Biol. Chem., 265, 18005–18011.

  • Craig AM, Bowden GT, Chambers AF, Spearman MA, Greenberg AH, Wright JA, McLeod M and Denhardt DT . (1990). Int. J. Cancer, 46, 133–137.

  • Craig AM, Smith JH and Denhardt DT . (1989). J. Biol. Chem., 264, 9682–9689.

  • Denhardt DT, Giachelli CM and Rittling SR . (2001a). Annu. Rev. Pharmacol. Toxicol., 41, 723–749.

  • Denhardt DT and Guo X . (1993). FASEB J., 7, 1475–1482.

  • Denhardt DT, Lopez CA, Rollo EE, Hwang SM, An XR and Walther SE . (1995). Ann. NY Acad. Sci., 760, 127142.

  • Denhardt DT, Mistretta D, Chambers AF, Krishna S, Porter JF, Raghuram S and Rittling SR . (2003). Clin. Exp. Metastasis, 20, 77–84.

  • Denhardt DT, Noda M, O’Regan AW, Pavlin D and Berman JS . (2001b). J. Clin. Invest., 107, 1055–1061.

  • Denko NC, Fontana LA, Hudson KM, Sutphin PD, Raychaudhuri S, Altman R and Giaccia AJ . (2003). Oncogene, 22, 5907–5914.

  • Ebert BL and Bunn HF . (1998). Mol. Cell. Biol., 18, 4089–4096.

  • Furger KA, Menon RK, Tuckl AB, Bramwelll VH and Chambers AF . (2001). Curr. Mol. Med., 1, 621–632.

  • Giaccia A, Siim BG and Johnson RS . (2003). Nat. Rev. Drug Discov., 2, 803–811.

  • Graeber TG, Osmanian C, Jacks T, Housman DE, Koch CJ, Lowe SW and Giaccia AJ . (1996). Nature, 379, 88–91.

  • Guo X, Zhang YP, Mitchell DA, Denhardt DT and Chambers AF . (1995). Mol. Cell. Biol., 15, 476–487.

  • Hammond EM, Denko NC, Dorie MJ, Abraham RT and Giaccia AJ . (2002). Mol. Cell. Biol., 22, 1834–1843.

  • Harris AL . (2002). Nat. Rev. Cancer, 2, 38–47.

  • Harrold S, Genovese C, Kobrin B, Morrison SL and Milcarek C . (1991). Anal. Biochem., 198, 19–29.

  • Hijiya N, Setoguchi M, Matsuura K, Higuchi Y, Akizuki S and Yamamoto S . (1994). Biochem. J., 303, 255–262.

  • Hockel M, Schlenger K, Aral B, Mitze M, Schaffer U and Vaupel P . (1996). Cancer Res., 56, 4509–4515.

  • Huang L, Zhang H, Bae S and Liu A . (1994). J. Biol. Chem., 269, 30718–30725.

  • Hwang SM, Wilson PD, Laskin JD and Denhardt DT . (1994). J. Cell Physiol., 160, 61–68.

  • Kohn AD, Summers SA, Birnbaum MJ and Roth RA . (1996). J. Biol. Chem., 271, 31372–31378.

  • Koong AC, Denko NC, Hudson KM, Schindler C, Swiersz L, Koch C, Evans S, Ibrahim H, Le QT, Terris DJ and Giaccia AJ . (2000). Cancer Res., 60, 883–887.

  • Koszewski NJ, Reinhardt TA and Horst RL . (1996). J. Steroid Biochem. Mol. Biol., 59, 377–388.

  • Le QT, Denko NC and Giaccia AJ . (2004). Cancer Metastasis Rev., 23, 293–310.

  • Le QT, Sutphin PD, Raychaudhuri S, Yu SC, Terris DJ, Lin HS, Lum B, Pinto HA, Koong AC and Giaccia AJ . (2003). Clin. Cancer Res., 9, 59–67.

  • Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM and Hogan BL . (1998). J. Clin. Invest., 101, 1468–1478.

  • Lowy DR and Willumsen BM . (1993). Annu. Rev. Biochem., 62, 851–891.

  • Mazure NM, Chen EY, Laderoute KR and Giaccia AJ . (1997). Blood, 90, 3322–3331.

  • Noda M, Vogel RL, Craig AM, Prahl J, DeLuca HF and Denhardt DT . (1990). Proc. Natl. Acad. Sci. USA, 87, 9995–9999.

  • Noiri E, Dickman K, Miller F, Romanov G, Romanov VI, Shaw R, Chambers AF, Rittling SR, Denhardt DT and Goligorsky MS . (1999). Kidney Int., 56, 74–82.

  • Pollmann C, Huang X, Mall J, Bech-Otschir D, Naumann M and Dubiel W . (2001). Cancer Res., 61, 8416–8421.

  • Semenza GL . (2003). Nat. Rev. Cancer, 3, 721–732.

  • Sodhi CP, Batlle D and Sahai A . (2000). Kidney Int., 58, 691–700.

  • Sodhi CP, Phadke SA, Batlle D and Sahai A . (2001). Diabetes, 50, 1482–1490.

  • Teicher BA, Holden SA, al-Achi A and Herman TS . (1990). Cancer Res., 50, 3339–3344.

  • Teramoto H, Malek RL, Behbahani B, Castellone MD, Lee NH and Gutkind JS . (2003). Oncogene, 22, 2689–2697.

  • Vaupel P, Thews O and Hoeckel M . (2001). Med. Oncol., 18, 243–259.

  • Weber GF . (2001). Biochim. Biophys. Acta., 1552, 61–85.

  • Weber GF and Cantor H . (1996). Cytokine Growth Factor Rev., 7, 241–248.

  • Xie Y, Sakatsume M, Nishi S, Narita I, Arakawa M and Gejyo F . (2001). Kidney Int., 60, 1645–1657.

  • Young SD and Hill RP . (1990). J. Natl. Cancer Inst., 82, 371–380.

  • Zhang G, He B and Weber GF . (2003). Mol. Cell. Biol., 23, 6507–6519.

  • Zhu Y, Hon T, Ye W and Zhang L . (2002a). Cell Growth Differ., 13, 431–439.

  • Zhu Y, Hon T and Zhang L . (1999). Biochem. Biophys. Res. Commun., 258, 87–93.

  • Zhu Y, Sun Y, Mao XO, Jin KL and Greenberg DA . (2002b). Neuroscience, 110, 191–198.

  • Zhu Y, Sun Y, Xie L, Jin K, Sheibani N and Greenberg DA . (2003). Stroke, 34, 2483–2488.

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Acknowledgements

This work was supported by the USPHS Grant CA 67166 (YZ, AJG, QTL) and the Damon Runyon–Lilly Clinical Investigator Award (ACK).

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Authors and Affiliations

  1. Department of Radiation Oncology, 875 Blake Wilbur Dr, R. CC-G228, Stanford University, Stanford, 94305, CA, USA

    Yonghua Zhu, Hongbin Cao, Patrick D Sutphin, Albert C Koong, Amato J Giaccia & Quynh-Thu Le

  2. Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, 08855, NJ, USA

    David T Denhardt

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  1. Yonghua Zhu
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Zhu, Y., Denhardt, D., Cao, H. et al. Hypoxia upregulates osteopontin expression in NIH-3T3 cells via a Ras-activated enhancer. Oncogene 24, 6555–6563 (2005). https://doi.org/10.1038/sj.onc.1208800

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