Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Transcriptional regulation of human osteopontin promoter by C/EBPα and AML-1 in metastatic cancer cells

Oncogene volume 23, pages 278–288 (2004)Cite this article

Abstract

Osteopontin (OPN) is a secreted glycoprotein produced by osteoclasts, macrophages, T cells, hematopoietic cells, and vascular smooth muscle cells. It contributes to macrophage homing and cellular immunity. It also mediates neovascularization, inhibits apoptosis, and plays important roles in extracellular matrix remodeling and angiogenesis. These properties are also characteristics of metastatic cancer cells. Consequently, the OPN gene was found to be upregulated among various metastatic cancer cells. This suggests that OPN is involved in tumor metastasis. How the OPN gene is upregulated in metastatic cancer cells remains to be illustrated. Thus, we investigated the transcriptional activation of the OPN promoter in the human metastatic cancer cell line A2058. We cloned the OPN promoter, and serial deletion analysis of the OPN promoter showed that the region between –170 and –127 may act as an enhancer to control the OPN gene in metastatic tumor cells. This region was found to contain overlapped AML-1 and C/EBP binding site motifs. Gel-mobility-shift assays using the A2058 nuclear extract and AML-1a or C/EBPα (CCAAT/enhancer binding protein alpha) recombinant protein indicated that these two transcription factors can bind to the overlapped AML-1 /C/EBP binding site motifs on the OPN regulatory sequence from –147 to –127. Surprisingly, the gel-shift experiments did not show supershift complex formation between AML-1 and C/EBPα. Functional analysis showed that the C/EBPα was more potent than the complex of AML-1 and its cofactor CBFβ to upregulate the OPN promoter. In addition, AML-1 and C/EBPα did not exhibit transactivation additively or synergistically. Our results suggest that AML-1 and C/EBPα play an important role in the upregulation of the OPN gene in metastatic tumor cells.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10
Figure 11

Similar content being viewed by others

References

  • Anna T, Antonietta RF, Ida V, Antonella T, Antonella T, Giancarlo S, Ann FC, Alberto G and Andrew RM . (1998). Int. J. Cancer, 77, 82–93.

  • Baiker A, Maercker C, Piechaczek C, Schmidt SBA, Bode J, Benham C and Lipps HJ . (2000). Nat. Cell Biol., 2, 182–184.

  • Brown LF, Papadopoulos SA, Berse B, Manseau EJ, Tognazzi K, Perruzzi CA, Dvorak HF and Senger DR . (1994). Am. J. of Pathol., 154, 610–623.

  • Chabas D, Baranzini SE, Mitchell D, Bernard CCA, Rittling SR, Denhardt DT, Sobel RA, Lock C, Karpuj M, Pedotti R, Heller R, Oksenberg JR and Steinman L . (2001). Science, 294, 1731–1735.

  • Chirgwin JM, Przybyla AE, MacDonald RJ and Rutter WJ . (1979). Biochemistry, 18, 5294–5299.

  • Crawford HC, Matrisian LM and Liaw L . (1998). Cancer Res., 58, 5206–5215.

  • Dipak PR and Pelagia F . (2002). Biochem. J., 365, 561–575.

  • Dongyan W, Shunsuke Y, Naoki H, Etty NB and Candece LG . (2000). Oncogene, 19, 5801–5809.

  • Fumiyuki T, Shigeru A, Hisao F, Shoji T, Yuichiro O, Kazuhisa T, Yoshinosuke F, Nagahiro S and Kazuto N . (2002). Int. J. Cancer, 98, 707–712.

  • Gang X, Ueki K, Kon S, Maeda M, Naruse T and Nojima Y . (2001). Am. J. Kidney Dis., 37, 374–379.

  • Guan KL and Dixon JE . (1991). Anal. Biochem., 192, 262–267.

  • Hanahan D and Weinberg RA . (2000). Cell, 100, 57–70.

  • Helmut P, Larry S and Peter AH . (2003). Nat. Rev. Mol. Cell. Biol., 4, 33–45.

  • John DD and Russell ML Robert GR . (1983). Nucleic Acids Res., 11, 1475–1489.

  • Kaartinen MT, Linnala-KA and Maenpaa PH . (1999). J. Biol. Chem., 274, 1729–1735.

  • Levanon D, Negreanu V, Bernstein Y, Bar-AI, Avivi L and Groner Y . (1994). Genomics, 23, 425–432.

  • Lisa MC and Zena W . (2002). Nature, 420, 860–867.

  • Liu P, Tarle SA, Hajra A, Claxton DF, Marlton P, Freedman M, Siciliano MJ and Collins FS . (1993). Science, 261, 1041–1044.

  • Lorsbach RB and Downing JR . (2001). Int. J. Hematol, 74, 258–265.

  • Lund AH and Van LM . (2002). Cancer Cell, 3, 213–215.

  • Meyers S, Downing JR and Hiebert SW . (1993). Mol. Cell. Biol., 13, 6336–6345.

  • Meyers S, Lenny N and Hiebert SW . (1995). Mol. Cell. Biol., 15, 1974–1982.

  • Mignatti P and Rifkin DB . (1996). Enzyme Protein, 49, 117–137.

  • Mikala E and Zena W . (2002). Nat. Rev. Cancer, 2, 163–176.

  • Miyoshi H, Ohira M, Shimizu K, Mitani K, Hirai H, Imai T, Yokoyama K, Soeda E and Ohki M . (1995). Nucleic Acids Res., 14, 2762–2769.

  • Nau GJ, Liaw L, Chupp GL, Berman JS, Hogan BL and Young RA . (1999). Infect. Immun., 67, 4223–4230.

  • Per A and Kleanthis GX . (1995). Biochem. Biophys. Res. Commun., 215, 106–113.

  • Petrovick MS, Hiebert SW, Friedman AD, Hetherington CJ, Tenen DG and Zhang DE . (1998). Mol. Cell. Biol., 18, 3915–3925.

  • Rittling SR, Chen Y, Feng F and Wu Y . (2002). J. Biol. Chem., 277, 9175–9182.

  • Rudland PS, Platt-Higgins A, El-Tanani M, De Silva RS, Barraclough R, Winstanley JH, Howitt R and West CR . (2002). Cancer Res., 62, 3417–3427.

  • Sato M, Morii E, Komori T, Kawahata H, Sugimoto M, Terai K, Shimizu H, Yasui T, Ogihara H, Yasui N, Ochi T, Kitamura Y, Ito Y and Nomura S . (1998). Oncogene, 17, 1517–1525.

  • Shi X, Bai S, Li L and Cao X . (2001). J Biol. Chem., 276, 850–855.

  • Sporn MB . (1996). Lancet, 347, 1377–1381.

  • Subha P, Anuradha B and Gopal CK . (2001). J. Biol. Chem., 276, 44926–44935.

  • Tomizawa M, Wang YQ, Ebara M, Saisho H, Watanabe K, Nakagawara A and Tagawa M . (2002). Int. J. Mol. Med., 9, 597–600.

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

  • Weber GF, Ashkar S, Glimcher MJ and Cantor H . (1996). Science, 271, 509–512.

  • Zhang DE, Hetherington CJ, Meyers S, Rhoades KL, Larson CJ, Chen HM, Hiebert SW and Tenen DG . (1996). Mol. Cell Biol., 16, 1231–1240.

Download references

Acknowledgements

We acknowledge the technical assistance of GJ Chen and CH Yuan, Aaron Lai for participation in purification of recombinant AML-1 and C/EBPα. We also thank Drs N Lenny, J Downing, H Miyoshi, and D Tenen for providing reagents, and members of Chen lab for discussions and suggestions. This work was supported by Grants NSC 91-2745-B-320-001 and NSC 91-23320-B-320-012 from National Science Council, Taiwan and Tzu Chi Foundation.

Author information

Authors and Affiliations

  1. Graduate Institute of Human Genetics, Tzu Chi University, Hualien, Taiwan

    Yan-Nien Liu & J H Chen

  2. Department of Medical Technology, College of Medicine, Tzu Chi University, Hualien, Taiwan

    Bean-Bu Kang

  3. Graduate Institute of Molecular and Cell Biology, Tzu Chi University, Hualien, Taiwan

    J H Chen

Authors
  1. Yan-Nien Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bean-Bu Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J H Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J H Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, YN., Kang, BB. & Chen, J. Transcriptional regulation of human osteopontin promoter by C/EBPα and AML-1 in metastatic cancer cells. Oncogene 23, 278–288 (2004). https://doi.org/10.1038/sj.onc.1207022

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207022

Keywords

This article is cited by

Search

Advanced search

Quick links