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:

Restoration of SMAD4 by gene therapy reverses the invasive phenotype in pancreatic adenocarcinoma cells

Oncogene volume 22, pages 6857–6864 (2003)Cite this article

Abstract

SMAD4 is a critical cofactor in signal transduction pathways activated in response to transforming growth factor-beta (TGF-β)-related ligands, regulating cell growth and differentiation. The roles played by SMAD4 inactivation in tumours highlighted it as a tumour-suppressor gene. However, restoration of the TGF-β antiproliferative pathway following SMAD4 gene transfer in null-tumour cell lines is controversial. Herein, we report the inhibitory effects of SMAD4 on pancreatic tumour invasion and angiogenesis. Adenoviral transfer of this gene in a panel of SMAD4 homozygous-deleted human pancreatic tumour cell lines restored SMAD4 protein expression and function. Although it did not affect proliferation significantly in vitro, SMAD4 inhibited in vivo tumour growth in immunodeficient mice. In this xenograft setting, differential suppression of tumour growth in vivo was mediated, at least in part, through downregulation of vascular endothelial growth factor and expression of gelatinases. We documented the reduced invasion and angiogenesis histologically and by intravital microscopy, and gained mechanistic insight at the messenger and protein level. Finally, we found a negative reciprocal regulation between SMAD4 and ETS-1. ETS-1 is considered a marker for tumour invasion. Upon SMAD4 deletion, we detected high expression levels of ETS-1 in pancreatic tumour cells, suggesting the shift of the pancreatic tumour toward an invasive phenotype.

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 4
Figure 2
Figure 3
Figure 5

Similar content being viewed by others

References

  • Attiga FA, Fernandez PM, Weeraratna AT, Manyak MJ and Patierno SR . (2000). Cancer Res., 60, 4629–4637.

  • Behrens P, Rothe M, Florin A, Wellmann A and Wernert N . (2001). Int. J. Mol. Med., 8, 149–154.

  • Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z and Hanahan D . (2000). Nat. Cell Biol., 2, 737–744.

  • Bramhall SR . (1997). Int. J. Pancreatol., 21, 1–12.

  • Calonge MJ and Massague J . (1999). J. Biol. Chem., 274, 33637–33643.

  • Dai JL, Bansal RK and Kern SE . (1999a). Proc. Natl. Acad. Sci. USA, 96, 1427–1432.

  • Dai JL, Schutte M, Bansal RK, Wilentz RE, Sugar AY and Kern SE . (1999b). Mol. Carcinog., 26, 37–43.

  • Davidson B, Reich R, Goldberg I, Gotlieb WH, Kopolovic J, Berner A, Ben-Baruch G, Bryne M and Nesland JM . (2001). Clin. Cancer Res., 7, 551–557.

  • Duda DG, Sunamura M, Lozonschi L, Kodama T, Egawa S, Matsumoto G, Shimamura H, Shibuya K, Takeda K and Matsuno S . (2000). Cancer Res., 60, 1111–1116.

  • Elvert G, Kappel A, Heidenreich R, Englmeier U, Lanz S, Acker T, Rauter M, Sieweke M, Breier G and Flamme I . (2003). J. Biol. Chem., 278, 7520–7530.

  • Eskinazi R, Resibois A, Svoboda M, Peny MO, Adler M, Robberecht P and Van Laethem JL . (1998). Gastroenterology, 114, 1211–1120.

  • Fink SP, Swinler SE, Lutterbaugh JD, Massague J, Thiagalingam S, Kinzler KW, Vogelstein B, Willson JK and Markowitz S . (2001). Cancer Res., 61, 256–260.

  • Friess H, Yamanaka Y, Buchler M, Ebert M, Beger HG, Gold LI and Korc M . (1993). Gastroenterology, 105, 1846–1856.

  • Fukushige S, Furukawa T, Satoh K, Sunamura M, Kobari M, Koizumi M and Horii A . (1998). Cancer Res., 58, 4222–4226.

  • Furukawa T, Duguid WP, Rosenberg L, Viallet J, Galloway DA and Tsao MS . (1996). Am. J. Pathol., 148, 1763–1770.

  • Inoue H, Furukawa T, Sunamura M, Takeda K, Matsuno S and Horii A . (2001). Genes Chromosomes Cancer, 31, 295–299.

  • Ito T, Nakayama T, Ito M, Naito S, Kanematsu T and Sekine I . (1998). Mod. Pathol., 11, 209–215.

  • Itoh T, Tanioka M, Yoshida H, Yoshioka T, Nishimoto H and Itohara S . (1998). Cancer Res., 58, 1048–1051.

  • Iwasaka C, Tanaka K, Abe M and Sato Y . (1996). J. Cell. Physiol., 169, 522–531.

  • Jiang Y, Liang H, Guo W, Kottickal LV and Nagarajan L . (2000). Blood, 95, 3945–3950.

  • Jiao S, Cheng L, Wolff JA and Yang NS . (1993). Biotechnology (N Y), 11, 497–502.

  • Kita D, Takino T, Nakada M, Takahashi T, Yamashita J and Sato H . (2001). Cancer Res., 61, 7985–7991.

  • Kondo E, Horii A and Fukushige S . (1999). J. Biochem. (Tokyo), 125, 818–825.

  • Lagna G, Hata A, Hemmati-Brivanlou A and Massague J . (1996). Nature, 383, 832–836.

  • Lefter LP, Furukawa T, Sunamura M, Duda DG, Takeda K, Kotobuki N, Oshimura M, Matsuno S and Horii A . (2002). Genes Chromosomes Cancer, 34, 234–242.

  • Nakayama T, Ito M, Ohtsuru A, Naito S and Sekine I . (2001). Mod. Pathol., 14, 415–422.

  • Nelson AR, Fingleton B, Rothenberg ML and Matrisian LM . (2000). J. Clin. Oncol., 18, 1135–1149.

  • Nishikawa A, Iwasaki M, Akutagawa N, Manase K, Yamashita S, Endo T and Kudo R . (2000). Gynecol. Oncol., 79, 256–263.

  • Oh SP, Seki T, Goss KA, Imamura T, Yi Y, Donahoe PK, Li L, Miyazono K, ten Dijke P, Kim S and Li E . (2000). Proc. Natl. Acad. Sci. USA, 97, 2626–2631.

  • Peng B, Fleming JB, Breslin T, Grau AM, Fojioka S, Abbruzzese JL, Evans DB, Ayers D, Wathen K, Wu T, Robertson KD and Chiao PJ . (2002). Clin. Cancer Res., 8, 3628–3638.

  • Ramachandra M, Atencio I, Rahman A, Vaillancourt M, Zou A, Avanzini J, Wills K, Bookstein R and Shabram P . (2002). Cancer Res., 62, 6045–6051.

  • Reisdorff J, En-Nia A, Stefanidis I, Floege J, Lovett DH and Mertens PR . (2002). J. Am. Soc. Nephrol., 13, 1568–1578.

  • Sato Y, Teruyama K, Nakano T, Oda N, Abe M, Tanaka K and Iwasaka-Yagi C . (2001). Ann. NY Acad. Sci., 947, 117–123.

  • Schwarte-Waldhoff I, Volpert OV, Bouck NP, Sipos B, Hahn SA, Klein-Scory S, Luttges J, Kloppel G, Graeven U, Eilert-Micus C, Hintelmann A and Schmiegel W . (2000). Proc. Natl. Acad. Sci. USA, 97, 9624–9629.

  • Span PN, Manders P, Heuvel JJ, Thomas CM, Bosch RR, Beex LV and Sweep CG . (2002). Oncogene, 21, 8506–8509.

  • St Croix B, Rago C, Velculescu V, Traverso G, Romans KE, Montgomery E, Lal A, Riggins GJ, Lengauer C, Vogelstein B and Kinzler KW . (2000). Science, 289, 1197–1202.

  • Sun C, Yamato T, Furukawa T, Ohnishi Y, Kijima H and Horii A . (2001). Oncol. Rep., 8, 89–92.

  • Sun Y, Wenger L, Rutter JL, Brinckerhoff CE and Cheung HS . (1999). J. Biol. Chem., 274, 11535–11540.

  • Takaku K, Oshima M, Miyoshi H, Matsui M, Seldin MF and Taketo MM . (1998). Cell, 92, 645–656.

  • Taniguchi N, Miyoshi E, Ko JH, Ikeda Y and Ihara Y . (1999). Biochim. Biophys. Acta., 1455, 287–300.

  • Teruyama K, Abe M, Nakano T, Iwasaka-Yagi C, Takahashi S, Yamada S and Sato Y . (2001). J. Cell Physiol., 188, 243–252.

  • Thant AA, Sein TT, Liu E, Machida K, Kikkawa F, Koike T, Seiki M, Matsuda S and Hamaguchi M . (1999). Oncogene, 18, 6555–6563.

  • Valter MM, Hugel A, Huang HJ, Cavenee WK, Wiestler OD, Pietsch T and Wernert N . (1999). Cancer Res., 59, 5608–5614.

  • Vlaykova T, Muhonen T, Hahka-Kemppinen M, Pyrhonen S and Jekunen A . (1997). Int. J. Cancer, 74, 326–329.

  • Vu TH, Shipley JM, Bergers G, Berger JE, Helms JA, Hanahan D, Shapiro SD, Senior RM and Werb Z . (1998). Cell, 93, 411–422.

  • Wilentz RE, Iacobuzio-Donahue CA, Argani P, McCarthy DM, Parsons JL, Yeo CJ, Kern SE and Hruban RH . (2000). Cancer Res., 60, 2002–2006.

  • Yang X, Castilla LH, Xu X, Li C, Gotay J, Weinstein M, Liu PP and Deng CX . (1999). Development, 126, 1571–1580.

  • Yatsuoka T, Sunamura M, Furukawa T, Fukushige S, Yokoyama T, Inoue H, Shibuya K, Takeda K, Matsuno S and Horii A . (2000). Am. J. Gastroenterol., 95, 2080–2085.

Download references

Acknowledgements

This work was supported in part by two grants-in-aid for scientific research (14657277) from the Japanese Ministry of Education, Culture, Sports, Science and Technology and grants-in-aid for cancer research from the Ministry of Health, Labour and Welfare of Japan. We thank Drs Kohei Miyazono (Department of Biochemistry, The Cancer Institute, Tokyo, Japan) and Yasufumi Sato (Department of Vascular Biology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan) for supplying the adenoviral constructs. We are grateful to Emiko Shibuya, Hiroko Fujimura, and Keiko Inabe at Tohoku University School of Medicine (Sendai, Japan) for their expert technical assistance.

Author information

Author notes

  1. Dan G Duda

    Present address: Steele Laboratory for Tumor Biology, Cox-734, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02214, USA

Authors and Affiliations

  1. Division of Gastroenterological Surgery, Graduate School of Medicine, Tohoku University, Sendai, 980-8574, Japan

    Dan G Duda, Makoto Sunamura, Liviu P Lefter, Tadaaki Yokoyama, Toshimasa Yatsuoka, Tadayoshi Abe, Hiroko Inoue, Fuyuhiko Motoi, Shin-ichi Egawa & Seiki Matsuno

  2. Department of Molecular Pathology, Graduate School of Medicine, Tohoku University, Sendai, 980-8575, Japan

    Toru Furukawa & Akira Horii

Authors
  1. Dan G Duda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Makoto Sunamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Liviu P Lefter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Toru Furukawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tadaaki Yokoyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Toshimasa Yatsuoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tadayoshi Abe
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hiroko Inoue
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Fuyuhiko Motoi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shin-ichi Egawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Seiki Matsuno
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Akira Horii
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Makoto Sunamura.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Duda, D., Sunamura, M., Lefter, L. et al. Restoration of SMAD4 by gene therapy reverses the invasive phenotype in pancreatic adenocarcinoma cells. Oncogene 22, 6857–6864 (2003). https://doi.org/10.1038/sj.onc.1206751

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1206751

Keywords

This article is cited by

Search

Advanced search

Quick links