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 Article
  • Published:

Runx3 is required for the differentiation of lung epithelial cells and suppression of lung cancer

Abstract

Human lung adenocarcinoma, the most prevalent form of lung cancer, is characterized by many molecular abnormalities. K-ras mutations are associated with the initiation of lung adenocarcinomas, but K-ras-independent mechanisms may also initiate lung tumors. Here, we find that the runt-related transcription factor Runx3 is essential for normal murine lung development and is a tumor suppressor that prevents lung adenocarcinoma. Runx3−/− mice, which die soon after birth, exhibit alveolar hyperplasia. Importantly, Runx3−/− bronchioli exhibit impaired differentiation, as evidenced by the accumulation of epithelial cells containing specific markers for both alveolar (that is SP-B) and bronchiolar (that is CC10) lineages. Runx3−/− epithelial cells also express Bmi1, which supports self-renewal of stem cells. Lung adenomas spontaneously develop in aging Runx3+/− mice (18 months after birth) and invariably exhibit reduced levels of Runx3. As K-ras mutations are very rare in these adenomas, Runx3+/− mice provide an animal model for lung tumorigenesis that recapitulates the preneoplastic stage of human lung adenocarcinoma development, which is independent of K-Ras mutation. We conclude that Runx3 is essential for lung epithelial cell differentiation, and that downregulation of Runx3 is causally linked to the preneoplastic stage of lung adenocarcinoma.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Bangsow C, Rubins N, Glusman G, Bernstein Y, Negreanu V, Goldenberg D et al. (2001). The RUNX3 gene—sequence, structure and regulated expression. Gene 279: 221–232.

    Article  CAS  PubMed  Google Scholar 

  • Bea S, Tort F, Pinyol M, Puig X, Hernandez L, Hernandez S et al. (2001). BMI-1 gene amplification and overexpression in hematological malignancies occur mainly in mantle cell lymphomas. Cancer Res 61: 2409–2412.

    CAS  PubMed  Google Scholar 

  • Belinsky SA, Nikula KJ, Palmisano WA, Michels R, Saccomanno G, Gabrielson E et al. (1998). Aberrant methylation of p16 (INK4a) is an early event in lung cancer and a potential biomarker for early diagnosis. Proc Natl Acad Sci USA 95: 11891–11896.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bishop AE . (2004). Pulmonary epithelial stem cells. Cell Prolif 37: 89–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cardoso WV, Lu J . (2006). Regulation of early lung morphogenesis: questions, facts and controversies. Development 133: 1611–1624.

    CAS  PubMed  Google Scholar 

  • Cazorla M, Hernandez L, Fernandez PL, Fabra A, Peinado MA, Dasenbrock C et al. (1998). Ki-ras gene mutations and absence of p53 gene mutations in spontaneous and urethane-induced early lung lesions in CBA/J mice. Mol Carcinog 21: 251–260.

    Article  CAS  PubMed  Google Scholar 

  • Chi XZ, Kim J, Lee YH, Lee JW, Lee KS, Wee H et al. (2009). Runt-related transcription factor RUNX3 is a target of MDM2-mediated ubiquitination. Cancer Res 69: 8111–8119.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chi XZ, Yang JO, Lee KY, Ito K, Sakakura C, Li QL et al. (2005). RUNX3 suppresses gastric epithelial cell growth by inducing p21(WAF1/Cip1) expression in cooperation with transforming growth factor {beta}-activated SMAD. Mol Cell Biol 25: 8097–8107.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Collins LG, Haines C, Perkel R, Enck RE . (2007). Lung cancer: diagnosis and management. Am Fam Physician 75: 56–63.

    PubMed  Google Scholar 

  • Costa RH, Kalinichenko VV, Lim L . (2001). Transcription factors in mouse lung development and function. Am J Physiol Lung Cell Mol Physiol 280: L823–L838.

    Article  CAS  PubMed  Google Scholar 

  • Dimri GP, Martinez JL, Jacobs JJ, Keblusek P, Itahana K, Van Lohuizen M et al. (2002). The Bmi-1 oncogene induces telomerase activity and immortalizes human mammary epithelial cells. Cancer Res 62: 4736–4745.

    CAS  PubMed  Google Scholar 

  • Dovey JS, Zacharek SJ, Kim CF, Lees JA . (2008). Bmi1 is critical for lung tumorigenesis and bronchioalveolar stem cell expansion. Proc Natl Acad Sci USA 105: 11857–11862.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fainaru O, Woolf E, Lotem J, Yarmus M, Brenner O, Goldenberg D et al. (2004). Runx3 regulates mouse TGF-beta-mediated dendritic cell function and its absence results in airway inflammation. EMBO J 23: 969–979.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goel A, Arnold CN, Tassone P, Chang DK, Niedzwiecki D, Dowell JM et al. (2004). Epigenetic inactivation of RUNX3 in microsatellite unstable sporadic colon cancers. Int J Cancer 112: 754–759.

    Article  CAS  PubMed  Google Scholar 

  • Guo WH, Weng LQ, Ito K, Chen LF, Nakanishi H, Tatematsu M et al. (2002). Inhibition of growth of mouse gastric cancer cells by Runx3, a novel tumor suppressor. Oncogene 21: 8351–8355.

    Article  CAS  PubMed  Google Scholar 

  • Inoue K, Ozaki S, Shiga T, Ito K, Masuda T, Okado N et al. (2002). Runx3 controls the axonal projection of proprioceptive dorsal root ganglion neurons. Nat Neurosci 5: 946–954.

    Article  CAS  PubMed  Google Scholar 

  • Ito K, Inoue KI, Bae SC, Ito Y . (2009). Runx3 expression in gastrointestinal tract epithelium: resolving the controversy. Oncogene 28: 1379–1384.

    Article  CAS  PubMed  Google Scholar 

  • Ito K, Lim AC, Salto-Tellez M, Motoda L, Osato M, Chuang LS et al. (2008). RUNX3 attenuates beta-catenin/T cell factors in intestinal tumorigenesis. Cancer Cell 14: 226–237.

    Article  CAS  PubMed  Google Scholar 

  • Ito K, Liu Q, Salto-Tellez M, Yano T, Tada K, Ida H et al. (2005). RUNX3, a novel tumor suppressor, is frequently inactivated in gastric cancer by protein mislocalization. Cancer Res 65: 7743–7750.

    Article  CAS  PubMed  Google Scholar 

  • Jackson EL, Willis N, Mercer K, Bronson RT, Crowley D, Montoya R et al. (2001). Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras. Genes Dev 15: 3243–3248.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kang GH, Lee S, Lee HJ, Hwang KS . (2004). Aberrant CpG island hypermethylation of multiple genes in prostate cancer and prostatic intraepithelial neoplasia. J Pathol 202: 233–240.

    Article  CAS  PubMed  Google Scholar 

  • Kato N, Tamura G, Fukase M, Shibuya H, Motoyama T . (2003). Hypermethylation of the RUNX3 gene promoter in testicular yolk sac tumor of infants. Am J Pathol 163: 387–391.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim CF, Jackson EL, Woolfenden AE, Lawrence S, Babar I, Vogel S et al. (2005a). Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell 121: 823–835.

    Article  CAS  PubMed  Google Scholar 

  • Kim WJ, Kim EJ, Jeong P, Quan C, Kim J, Li QL et al. (2005b). RUNX3 inactivation by point mutations and aberrant DNA methylation in bladder tumors. Cancer Res 65: 9347–9354.

    Article  CAS  PubMed  Google Scholar 

  • Lau QC, Raja E, Salto-Tellez M, Liu Q, Ito K, Inoue M et al. (2006). RUNX3 is frequently inactivated by dual mechanisms of protein mislocalization and promoter hypermethylation in breast cancer. Cancer Res 66: 6512–6520.

    Article  CAS  PubMed  Google Scholar 

  • Lee B, Thirunavukkarasu K, Zhou L, Pastore L, Baldini A, Hecht J et al. (1997). Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFA1 in cleidocranial dysplasia. Nat Genet 16: 307–310.

    Article  CAS  PubMed  Google Scholar 

  • Leung C, Lingbeek M, Shakhova O, Liu J, Tanger E, Saremaslani P et al. (2004). Bmi1 is essential for cerebellar development and is overexpressed in human medulloblastomas. Nature 428: 337–341.

    Article  CAS  PubMed  Google Scholar 

  • Levanon D, Bettoun D, Harris-Cerruti C, Woolf E, Negreanu V, Eilam R et al. (2002). The Runx3 transcription factor regulates development and survival of TrkC dorsal root ganglia neurons. EMBO J 21: 3454–3463.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li QL, Ito K, Sakakura C, Fukamachi H, Inoue K, Chi XZ et al. (2002). Causal relationship between the loss of RUNX3 expression and gastric cancer. Cell 109: 113–124.

    Article  CAS  PubMed  Google Scholar 

  • Li QL, Kim HR, Kim WJ, Choi JK, Lee YH, Kim HM et al. (2004). Transcriptional silencing of the RUNX3 gene by CpG hypermethylation is associated with lung cancer. Biochem Biophys Res Commun 314: 223–228.

    Article  CAS  PubMed  Google Scholar 

  • Licchesi JD, Westra WH, Hooker CM, Machida EO, Baylin SB, Herman JG . (2008). Epigenetic alteration of Wnt pathway antagonists in progressive glandular neoplasia of the lung. Carcinogenesis 29: 895–904.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Look AT . (1997). Oncogenic transcription factors in the human acute leukemias. Science 278: 1059–1064.

    Article  CAS  PubMed  Google Scholar 

  • Lund AH, van Lohuizen M . (2002). RUNX: a trilogy of cancer genes. Cancer Cell 1: 213–215.

    Article  CAS  PubMed  Google Scholar 

  • Malkinson AM . (1998). Molecular comparison of human and mouse pulmonary adenocarcinomas. Exp Lung Res 24: 541–555.

    Article  CAS  PubMed  Google Scholar 

  • Minna JD, Roth JA, Gazdar AF . (2002). Focus on lung cancer. Cancer Cell 1: 49–52.

    Article  CAS  PubMed  Google Scholar 

  • Mori M, Kaji M, Tezuka F, Takahashi T . (1998). Comparative ultrastructural study of atypical adenomatous hyperplasia and adenocarcinoma of the human lung. Ultrastruct Pathol 22: 459–466.

    Article  CAS  PubMed  Google Scholar 

  • Mundlos S, Olsen BR . (1997). Heritable diseases of the skeleton Part I: molecular insights into skeletal development-transcription factors and signaling pathways. FASEB J 11: 125–132.

    Article  CAS  PubMed  Google Scholar 

  • Okuda T, Fisher R, Downing JR . (1996). Molecular diagnostics in pediatric acute lymphoblastic leukemia. Mol Diagn 1: 139–151.

    Article  CAS  PubMed  Google Scholar 

  • Osanai M, Igarashi T, Yoshida Y . (2001). Unique cellular features in atypical adenomatous hyperplasia of the lung: ultrastructural evidence of its cytodifferentiation. Ultrastruct Pathol 25: 367–373.

    Article  CAS  PubMed  Google Scholar 

  • Otto F, Thornell AP, Crompton T, Denzel A, Gilmour KC, Rosewell IR et al. (1997). Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell 89: 765–771.

    Article  CAS  PubMed  Google Scholar 

  • Peacock CD, Watkins DN . (2008). Cancer stem cells and the ontogeny of lung cancer. J Clin Oncol 26: 2883–2889.

    Article  CAS  PubMed  Google Scholar 

  • Petersen I, Petersen S . (2001). Towards a genetic-based classification of human lung cancer. Anal Cell Pathol 22: 111–121.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Reddy R, Buckley S, Doerken M, Barsky L, Weinberg K, Anderson KD et al. (2004). Isolation of a putative progenitor subpopulation of alveolar epithelial type 2 cells. Am J Physiol Lung Cell Mol Physiol 286: L658–L667.

    Article  CAS  PubMed  Google Scholar 

  • Sato K, Tomizawa Y, Iijima H, Saito R, Ishizuka T, Nakajima T et al. (2006). Epigenetic inactivation of the RUNX3 gene in lung cancer. Oncol Rep 15: 129–135.

    CAS  PubMed  Google Scholar 

  • Taniuchi I, Sunshine MJ, Festenstein R, Littman DR . (2002). Evidence for distinct CD4 silencer functions at different stages of thymocyte differentiation. Mol Cell 10: 1083–1096.

    Article  CAS  PubMed  Google Scholar 

  • Tozawa T, Tamura G, Honda T, Nawata S, Kimura W, Makino N et al. (2004). Promoter hypermethylation of DAP-kinase is associated with poor survival in primary biliary tract carcinoma patients. Cancer Sci 95: 736–740.

    Article  CAS  PubMed  Google Scholar 

  • Tuveson DA, Shaw AT, Willis NA, Silver DP, Jackson EL, Chang S et al. (2004). Endogenous oncogenic K-ras(G12D) stimulates proliferation and widespread neoplastic and developmental defects. Cancer Cell 5: 375–387.

    Article  CAS  PubMed  Google Scholar 

  • Valk-Lingbeek ME, Bruggeman SW, van Lohuizen M . (2004). Stem cells and cancer; the polycomb connection. Cell 118: 409–418.

    Article  CAS  PubMed  Google Scholar 

  • Vonlanthen S, Heighway J, Altermatt HJ, Gugger M, Kappeler A, Borner MM et al. (2001). The bmi-1 oncoprotein is differentially expressed in non-small cell lung cancer and correlates with INK4A-ARF locus expression. Br J Cancer 84: 1372–1376.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Weisenberger DJ, Siegmund KD, Campan M, Young J, Long TI, Faasse MA et al. (2006). CpG island methylator phenotype underlies sporadic microsatellite instability and is tightly associated with BRAF mutation in colorectal cancer. Nat Genet 38: 787–793.

    Article  CAS  PubMed  Google Scholar 

  • Westra WH, Baas IO, Hruban RH, Askin FB, Wilson K, Offerhaus GJ et al. (1996). K-ras oncogene activation in atypical alveolar hyperplasias of the human lung. Cancer Res 56: 2224–2228.

    CAS  PubMed  Google Scholar 

  • Wistuba II, Gazdar AF . (2006). Lung cancer preneoplasia. Annu Rev Pathol 1: 331–348.

    Article  CAS  PubMed  Google Scholar 

  • Woolf E, Xiao C, Fainaru O, Lotem J, Rosen D, Negreanu V et al. (2003). Runx3 and Runx1 are required for CD8T cell development during thymopoiesis. Proc Natl Acad Sci USA 100: 7731–7736.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wuenschell CW, Sunday ME, Singh G, Minoo P, Slavkin HC, Warburton D . (1996). Embryonic mouse lung epithelial progenitor cells co-express immunohistochemical markers of diverse mature cell lineages. J Histochem Cytochem 44: 113–123.

    Article  CAS  PubMed  Google Scholar 

  • Xiao WH, Liu WW . (2004). Hemizygous deletion and hypermethylation of RUNX3 gene in hepatocellular carcinoma. World J Gastroenterol 10: 376–380.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yano T, Ito K, Fukamachi H, Chi XZ, Wee HJ, Inoue K et al. (2006). The RUNX3 tumor suppressor upregulates Bim in gastric epithelial cells undergoing transforming growth factor beta-induced apoptosis. Mol Cell Biol 26: 4474–4488.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zochbauer-Muller S, Minna JD . (2000). The biology of lung cancer including potential clinical applications. Chest Surg Clin N Am 10: 691–708.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by research grants from the Korea Science and Engineering Foundation (R16-2003–002–01001–02006 to S-C Bae and R13-2003–013–05001–0 to H-S Jung) and a grant from the Basic Research Promotion Fund of the Korea Research Foundation (KRF-2005–217-E00002 to K-S Lee).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to K-S Lee or S-C Bae.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, KS., Lee, YS., Lee, JM. et al. Runx3 is required for the differentiation of lung epithelial cells and suppression of lung cancer. Oncogene 29, 3349–3361 (2010). https://doi.org/10.1038/onc.2010.79

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2010.79

Keywords

This article is cited by

Search

Quick links