RUNX3 Downregulation in Human Lung Adenocarcinoma is Independent of p53, EGFR or KRAS Status

Mohd Feroz Mohd Omar; Kosei Ito; Min En Nga; Ross Soo; Bee Keow Peh; Tuty Muliana Ismail; Bhavin Thakkar; Richie Soong; Yoshiaki Ito; Manuel Salto-Tellez

doi:10.1007/s12253-011-9485-5

Pathology & Oncology Research

October 2012, Volume 18, Issue 4, pp 783–792

RUNX3 Downregulation in Human Lung Adenocarcinoma is Independent of p53, EGFR or KRAS Status

Authors
Authors and affiliations

Mohd Feroz Mohd Omar
Kosei Ito
Min En Nga
Ross Soo
Bee Keow Peh
Tuty Muliana Ismail
Bhavin Thakkar
Richie Soong
Yoshiaki ItoEmail author
Manuel Salto-TellezEmail author

Research

First Online:: 24 June 2012

Received:: 30 November 2010
Accepted:: 24 November 2011

DOI: 10.1007/s12253-011-9485-5

Cite this article as:: Omar, M.F.M., Ito, K., Nga, M.E. et al. Pathol. Oncol. Res. (2012) 18: 783. doi:10.1007/s12253-011-9485-5

Abstract

RUNX3 aberrations play a pivotal role in the oncogenesis of breast, gastric, colon, skin and lung tissues. The aim of this study was to characterize further the expression of RUNX3 in lung cancers. To achieve this, a lung cancer tissue microarray (TMA), frozen lung cancer tissues and lung cell lines were examined for RUNX3 expression by immunohistochemistry, while the TMA was also examined for EGFR and p53 expression. RUNX3 promoter methylation status, and EGFR and KRAS mutation status were also investigated. Inactivation of RUNX3 was observed in 70% of the adenocarcinoma samples, and this was associated with promoter hypermethylation but not biased to EGFR/KRAS mutations. Our results suggest a central role of RUNX3 downregulation in pulmonary adenocarcinoma, which may not be dependent of other established cancer-causing pathways and may have important diagnostic and screening implications.

Keywords

EGFR KRAS Lung adenocarcinoma p53 Promoter hypermethylation RUNX3

References

1.
Brambilla E, Travis WD, Colby TV, Corrin B, Shimosato Y (2001) The new World Health Organization classification of lung tumours. Eur Respir J 18(6):1059–1068PubMedCrossRefGoogle Scholar
2.
Mattson ME, Pollack ES, Cullen JW (1987) What are the odds that smoking will kill you? Am J Public Health 77(4):425–431PubMedCrossRefGoogle Scholar
3.
Hessel PA, Gamble JF, McDonald JC (2005) Asbestos, asbestosis, and lung cancer: a critical assessment of the epidemiological evidence. Thorax 60(5):433–436PubMedCrossRefGoogle Scholar
4.
Lubin JH, Boice JD Jr (1997) Lung cancer risk from residential radon: meta-analysis of eight epidemiologic studies. J Natl Cancer Inst 89(1):49–57PubMedCrossRefGoogle Scholar
5.
Zhu CQ, Shih W, Ling CH, Tsao MS (2006) Immunohistochemical markers of prognosis in non-small cell lung cancer: a review and proposal for a multiphase approach to marker evaluation. J Clin Pathol 59(8):790–800PubMedCrossRefGoogle Scholar
6.
Belinsky SA, Nikula KJ, Palmisano WA, Michels R, Saccomanno G, Gabrielson E et al (1998) Aberrant methylation of p16INK4a is an early event in lung cancer and a potential biomarker for early diagnosis. Proc Natl Acad Sci 95(20):11891–11896PubMedCrossRefGoogle Scholar
7.
Bouchardy C, Benhamou S, Dayer P (1996) The effect of tobacco on lung cancer risk depends on CYP2D6 activity. Cancer Res 56(2):251–253PubMedGoogle Scholar
8.
El-Zein RA, Schabath MB, Etzel CJ, Lopez MS, Franklin JD, Spitz MR (2006) Cytokinesis-blocked micronucleus assay as a novel biomarker for lung cancer risk. Cancer Res 66(12):6449–6456PubMedCrossRefGoogle Scholar
9.
Marchand LL, Sivaraman L, Pierce L, Seifried A, Lum A, Wilkens LR et al (1998) Associations of CYP1A1, GSTM1, and CYP2E1 polymorphisms with lung cancer suggest cell type specificities to tobacco carcinogens. Cancer Res 58(21):4858–4863PubMedGoogle Scholar
10.
Schwartz AG, Prysak GM, Bock CH, Cote ML (2007) The molecular epidemiology of lung cancer. Carcinogenesis 28(3):507–518PubMedCrossRefGoogle Scholar
11.
Sellers TA, Potter JD, Bailey-Wilson JE, Rich SS, Rothschild H, Elston RC (1992) Lung Cancer Detection and Prevention: Evidence for an Interaction between Smoking and Genetic Predisposition. Cancer Res. 52(9_Supplement):2694s-7Google Scholar
12.
Shaw GL, Falk RT, Frame JN, Weiffenbach B, Nesbitt JC, Pass HI et al (1998) Genetic polymorphism of CYP2D6 and lung cancer risk. Cancer Epidemiol Biomarkers Prev 7(3):215–219PubMedGoogle Scholar
13.
Wistuba II, Mao L, Gazdar AF (2002) Smoking molecular damage in bronchial epithelium. Oncogene 21(48):7298–7306PubMedCrossRefGoogle Scholar
14.
Yamabuki T, Takano A, Hayama S, Ishikawa N, Kato T, Miyamoto M et al (2007) Dikkopf-1 as a novel serologic and prognostic biomarker for lung and esophageal carcinomas. Cancer Res 67(6):2517–2525PubMedCrossRefGoogle Scholar
15.
Arenberg D (2004) In search of the holy grail: lung cancer biomarkers. Chest 126(2):325–326PubMedCrossRefGoogle Scholar
16.
Altshuler D, Daly MJ, Lander ES (2008) Genetic mapping in human disease. Science 322(5903):881–888PubMedCrossRefGoogle Scholar
17.
Duffy MJ, Crown J (2008) A personalized approach to cancer treatment: how biomarkers can help. Clin Chem 54(11):1770–1779PubMedCrossRefGoogle Scholar
18.
Brambilla C, Fievet F, Jeanmart M, de Fraipont F, Lantuejoul S, Frappat V et al (2003) Early detection of lung cancer: role of biomarkers. Eur Respir J. 21(39_suppl):36S-44Google Scholar
19.
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(2):221–232PubMedCrossRefGoogle Scholar
20.
Ito Y (2008) RUNX genes in development and cancer: regulation of viral gene expression and the discovery of RUNX family genes. Adv Cancer Res 99:33–76PubMedCrossRefGoogle Scholar
21.
Hanai J-i, Chen LF, Kanno T, Ohtani-Fujita N, Kim WY, Guo W-H et al (1999) Interaction and Functional Cooperation of PEBP2/CBF with Smads. Synergistic induction of the immunoglobulin germline Cα promoter. J Biol Chem 274(44):31577–31582PubMedCrossRefGoogle Scholar
22.
Ito K, Lim AC-B, Salto-Tellez M, Motoda L, Osato M, Chuang LSH et al (2008) RUNX3 attenuates β-Catenin/T cell factors in intestinal tumorigenesis. Cancer Cell 14(3):226–237PubMedCrossRefGoogle Scholar
23.
Licchesi JDF, 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(5):895–904PubMedCrossRefGoogle Scholar
24.
Homma N, Tamura G, Honda T, Matsumoto Y, Nishizuka S, Kawata S et al (2006) Spreading of methylation within RUNX3 CpG island in gastric cancer. Cancer Sci 97(1):51–56PubMedCrossRefGoogle Scholar
25.
Lee JM, Kwon HJ, Bae SC, Jung HS (2010) Lung tissue regeneration after induced injury in Runx3 KO mice. Cell Tissue Res 341(3):465–470PubMedCrossRefGoogle Scholar
26.
Lee KS, Lee YS, Lee JM, Ito K, Cinghu S, Kim JH et al (2010) Runx3 is required for the differentiation of lung epithelial cells and suppression of lung cancer. Oncogene 29(23):3349–3361PubMedCrossRefGoogle Scholar
27.
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(13):6512–6520PubMedCrossRefGoogle Scholar
28.
Subramaniam MM, Chan JY, Soong R, Ito K, Ito Y, Yeoh KG et al (2008) RUNX3 inactivation by frequent promoter hypermethylation and protein mislocalization constitute an early event in breast cancer progression. Breast Cancer Res Treat
29.
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(17):7743–7750PubMedGoogle Scholar
30.
Ku J-L, Kang S-B, Shin Y-K, Kang HC, Hong S-H, Kim I-J et al (2004) Promoter hypermethylation downregulates RUNX3 gene expression in colorectal cancer cell lines. Oncogene 23(40):6736–6742PubMedCrossRefGoogle Scholar
31.
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(1):113–124PubMedCrossRefGoogle Scholar
32.
Salto-Tellez M, Peh BK, Ito K, Tan SH, Chong PY, Han HC et al (2006) RUNX3 protein is overexpressed in human basal cell carcinomas. Oncogene 25(58):7646–7649PubMedCrossRefGoogle Scholar
33.
Kim TY, Lee HJ, Hwang KS, Lee M, Kim JW, Bang YJ et al (2004) Methylation of RUNX3 in various types of human cancers and premalignant stages of gastric carcinoma. Lab Invest 84(4):479–484PubMedCrossRefGoogle Scholar
34.
Yanada M, Yaoi T, Shimada J, Sakakura C, Nishimura M, Ito K et al (2005) Frequent hemizygous deletion at 1p36 and hypermethylation downregulate RUNX3 expression in human lung cancer cell lines. Oncol Rep 14(4):817–822PubMedGoogle Scholar
35.
Lane D (2004) p53 from pathway to therapy. Carcinogenesis 25(7):1077–1081PubMedCrossRefGoogle Scholar
36.
Cheng Y-L, Lee S-C, Harn H-J, Chen C-J, Chang Y-C, Chen J-C et al (2003) Prognostic prediction of the immunohistochemical expression of p53 and p16 in resected non-small cell lung cancer. Eur J Cardiothorac Surg 23(2):221–228PubMedCrossRefGoogle Scholar
37.
Maniwa Y, Yoshimura M, Obayashi C, Inaba M, Kiyooka K, Kanki M et al (2001) Association of p53 gene mutation and telomerase activity in resectable non-small cell lung cancer*. Chest 120(2):589–594PubMedCrossRefGoogle Scholar
38.
Piyathilake CJ, Frost AR, Manne U, Weiss H, Heimburger DC, Grizzle WE (2003) Nuclear accumulation of p53 is a potential marker for the development of squamous cell lung cancer in smokers*. Chest 123(1):181–186PubMedCrossRefGoogle Scholar
39.
Ettinger DS (2006) Clinical implications of EGFR expression in the development and progression of solid tumors: focus on non-small cell lung cancer. Oncologist 11(4):358–373PubMedCrossRefGoogle Scholar
40.
Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW et al (2004) Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N Engl J Med 350(21):2129–2139PubMedCrossRefGoogle Scholar
41.
Jang TW, Oak CH, Chang HK, Suo SJ, Jung MH (2009) EGFR and KRAS mutations in patients with adenocarcinoma of the lung. Korean J Intern Med 24(1):48–54PubMedCrossRefGoogle Scholar
42.
Shigematsu H, Lin L, Takahashi T, Nomura M, Suzuki M, Wistuba II et al (2005) Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers. J Natl Cancer Inst 97(5):339–346PubMedCrossRefGoogle Scholar
43.
Schmid K, Oehl N, Wrba F, Pirker R, Pirker C, Filipits M (2009) EGFR/KRAS/BRAF mutations in primary lung adenocarcinomas and corresponding locoregional lymph node metastases. Clin Cancer Res 15(14):4554–4560PubMedCrossRefGoogle Scholar
44.
Eberhard DA, Johnson BE, Amler LC, Goddard AD, Heldens SL, Herbst RS et al (2005) Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib. J Clin Oncol 23(25):5900–5909PubMedCrossRefGoogle Scholar
45.
Tsunematsu T, Kudo Y, Iizuka S, Ogawa I, Fujita T, Kurihara H et al (2009) RUNX3 has an oncogenic role in head and neck cancer. PLoS One 4(6):e5892PubMedCrossRefGoogle Scholar
46.
Yanagawa N, Tamura G, Oizumi H, Endoh M, Sadahiro M, Motoyama T (2011) Inverse correlation between EGFR mutation and FHIT, RASSF1A and RUNX3 methylation in lung adenocarcinoma: relation with smoking status. Anticancer Res 31(4):1211–1214PubMedGoogle Scholar
47.
Das K, Mohd Omar MF, Ong CW, Bin Abdul Rashid S, Peh BK, Putti TC et al (2008) TRARESA: a tissue microarray-based hospital system for biomarker validation and discovery. Pathology 40(5):441–449PubMedCrossRefGoogle Scholar
48.
Yano T, Ito K, Fukamachi H, Chi X-Z, Wee H-J, Inoue K-i et al (2006) The RUNX3 tumor suppressor upregulates bim in gastric epithelial cells undergoing transforming growth factor β-induced apoptosis. Mol Cell Biol 26(12):4474–4488PubMedCrossRefGoogle Scholar
49.
Soong R, Robbins PD, Dix BR, Grieu F, Lim B, Knowles S et al (1996) Concordance between p53 protein overexpression and gene mutation in a large series of common human carcinomas. Hum Pathol 27(10):1050–1055PubMedCrossRefGoogle Scholar
50.
Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB (1996) Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA 93(18):9821–9826PubMedCrossRefGoogle Scholar
51.
Krypuy M, Newnham G, Thomas D, Conron M, Dobrovic A (2006) High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer. BMC Cancer 6(1):295PubMedCrossRefGoogle Scholar
52.
Chin TM, Anuar D, Soo R, Salto-Tellez M, Li WQ, Ahmad B et al (2007) Detection of epidermal growth factor receptor variations by partially denaturing HPLC. Clin Chem 53(1):62–70PubMedCrossRefGoogle Scholar
53.
Araki K, Osaki M, Nagahama Y, Hiramatsu T, Nakamura H, Ohgi S et al (2005) Expression of RUNX3 protein in human lung adenocarcinoma: implications for tumor progression and prognosis. Cancer Sci 96(4):227–231PubMedCrossRefGoogle Scholar
54.
Ito K, Inoue Ki, Bae SC, Ito Y (2009) Runx3 expression in gastrointestinal tract epithelium: resolving the controversy. Oncogene 28(10):1379–1384PubMedCrossRefGoogle Scholar
55.
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(1):223–228PubMedCrossRefGoogle Scholar
56.
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(1):129–135PubMedGoogle Scholar
57.
Yanagawa N, Tamura G, Oizumi H, Takahashi N, Shimazaki Y, Motoyama T (2003) Promoter hypermethylation of tumor suppressor and tumor-related genes in non-small cell lung cancers. Cancer Sci 94(7):589–592PubMedCrossRefGoogle Scholar
58.
Ibrahim AEK, Arends MJ, Silva A-L, Wyllie AH, Greger L, Ito Y et al (2010) Sequential DNA methylation changes are associated with DNMT3B overexpression in colorectal neoplastic progression. Gut
59.
Soung YH, Lee JW, Kim SY, Seo SH, Park WS, Nam SW et al (2005) Mutational analysis of EGFR and K-RAS genes in lung adenocarcinomas. Virchows Arch 446(5):483–488PubMedCrossRefGoogle Scholar

Copyright information

Authors and Affiliations

Mohd Feroz Mohd Omar
- 1
- 2
Kosei Ito
- 3
Min En Nga
- 4
Ross Soo
- 1
- 5
Bee Keow Peh
- 1
- 4
Tuty Muliana Ismail
- 4
- 6
Bhavin Thakkar
- 1
Richie Soong
- 1
- 4
Yoshiaki Ito
- 1
- 2
Email author
Manuel Salto-Tellez
- 1
- 6
- 7
Email author

1.Cancer Science Institute of SingaporeCentre for Translational Medicine, National University of SingaporeSingaporeSingapore
2.NUS Graduate School for Integrative Sciences & EngineeringNational University of SingaporeSingaporeSingapore
3.Nagasaki University Graduate School of Biomedical ScienceNagasakiJapan
4.Department of PathologyNational University HospitalSingaporeSingapore
5.Department of Hematology-OncologyNational University HospitalSingaporeSingapore
6.Diagnostic Molecular Oncology Centre, #03-06, Clinical Research Centre, MD 11National University of SingaporeSingaporeSingapore
7.Centre for Cancer Research and Cell BiologyQueen’s University BelfastBT9 7BL BelfastUnited Kingdom

Personalised recommendations

Article actions

Unlimited access to the full article
Instant download
Include local sales tax if applicable

Subscribe to Journal

Get Access to

Pathology & Oncology Research

for the whole of 2017

RUNX3 Downregulation in Human Lung Adenocarcinoma is Independent of p53, EGFR or KRAS Status

Abstract

Keywords

Copyright information

Authors and Affiliations

Personalised recommendations

Cookies