Share This Article:

Promoter Methylation of the CADM1 and 4.1B Genes Occurs Independently of the EGFR or the KRAS2 Mutation in Non-Small Cell Lung Cancer

Abstract Full-Text HTML XML Download Download as PDF (Size:469KB) PP. 273-285
DOI: 10.4236/jct.2015.63030    2,808 Downloads   3,279 Views  


Objective: Targeting mutated EGFR by EGFR-tyrosine kinase inhibitors (EGFR-TKI) is a potent approach to a subset of non-small cell lung cancer (NSCLC). However, the response to EGFR-TKI varies in individual cases even among tumors carrying the same EGFR mutation, suggesting the involvement of modifying factors. To characterize possible modifiers, we examined mutation state of the EGFR and the KRAS genes in Japanese NSCLC and compared them with the methylation state of lung tumor suppressors, the CADM1 and 4.1B, whose products have potentials to modify the functions of EGFR or KRAS. Materials and methods: A total of 103 Japanese NSCLC and 11 NSCLC cell lines were examined. Genomic DNA of exons 18–21 of the EGFR and exons 1 and 2 of the KRAS were amplified by polymerase chain reaction (PCR), followed by single-strand conformation polymorphism analysis and direct sequencing. Methylation status of gene promoters in NSCLC cells were examined by methylation-specific PCR. Results: Mutations of the EGFR and KRAS were detected mutually exclusively in 27 and 11 out of 103 NSCLC cases, respectively. EGFR mutations were observed exclusively in adenocarcinoma (27 of 69, 41%) and preferentially in tumors from female and non-smokers (p < 0.00001). Eight (30%) and 12 (44%) of 27 tumors carrying mutated EGFR and 4 (36%) and 8 (73%) of 11 tumors carrying mutated KRAS showed methylation of the CADM1 and 4.1B, respectively. EGFR-mutated tumors with methylation of either CADM1 or 4.1showed more malignant features than those with unmethylated CADM1 and 4.1B (p < 0.05). Conclusion: Methylation state of the CADM1 and 4.1B are independent of the mutation status of the EGFR or KRAS but play roles in the malignant progression of NSCLC. Integration of epigenetic information would be useful for identifying possible modifiers to predict the response or recurrence of lung adenocarcinoma to the EGFR-TKI therapy.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Kogai, H. , Kikuchi, S. , Obana, T. , Tsuboi, Y. , Maruyama, T. , Sakurai-Yageta, M. , Asamura, H. , Kanai, Y. and Murakami, Y. (2015) Promoter Methylation of the CADM1 and 4.1B Genes Occurs Independently of the EGFR or the KRAS2 Mutation in Non-Small Cell Lung Cancer. Journal of Cancer Therapy, 6, 273-285. doi: 10.4236/jct.2015.63030.


[1] Minna, J.D., Roth, J.A. and Gazdar, A.F. (2002) Focus on Lung Cancer. Cancer Cell, 1, 49-52.
[2] Yokota, J. and Kohno, T. (2004) Molecular Footprints of Human Lung Cancer Progression. Cancer Sci, 95, 197-204.
[3] Mitsudomi, T. and Yatabe, Y. (2007) Mutations of the Epidermal Growth Factor Receptor Gene and Related Genes as Determinants of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors Sensitivity in Lung Cancer. Cancer Science, 98, 1817-1824.
[4] Soda, M., Choi, Y.L., Enomoto, M., Takada, S., Yamashita, Y., Ishikawa, S., Fujiwara, S., Watanabe, H., Kurashina, K., Hatanaka, H., Bando, M., Ohno, S., Ishikawa, Y., Aburatani, H., Niki, T., Sohara, Y., Sugiyama, Y. and Mano, H. (2007) Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer. Nature, 448, 561-566.
[5] Siddiqui, A.D. and Piperdi, B. (2010) KRAS Mutation in Colon Cancer: A Marker of Resistance to EGFR-I Therapy. Annals of Surgical Oncology, 17, 1168-1176.
[6] Bivona, T.G., Hieronymus, H., Parker, J., Chang, K., Taron, M., Rosell, R., Moonsamy, P., Dahlman, K., Miller, V.A., Costa, C., Hannon, G. and Sawyers, C.L. (2011) FAS and NF-κB Signalling Modulate Dependence of Lung Cancers on Mutant EGFR. Nature, 471, 523-526.
[7] Kuramochi, M., Fukuhara, H., Nobukuni, T., Kanbe, T., Maruyama, T., Ghosh, H.P., Pletcher, M., Isomura, M., Onizuka, M., Kitamura, T., Sekiya, T., Reeves, R.H. and Murakami, Y. (2001) TSLC1 Is a Tumor-Suppressor Gene in Human Non-Small-Cell Lung Cancer. Nature Genetics, 27, 427-430.
[8] Yageta, M., Kuramochi, M., Masuda, M., Fukami, T., Fukuhara, H., Maruyama, T., Shibuya, M. and Murakami, Y. (2002) Direct Association of TSLC1 and DAL-1, Two Distinct Tumor Suppressor Proteins in Lung Cancer. Cancer Research, 62, 5129-5133.
[9] Tran, Y.K., Bogler, O., Gorse, K.M., Wieland, I. and Green, M.R. (1999) A Novel Member of the NF2/ERM/4.1 Superfamily with Growth Suppressing Properties in Lung Cancer. Cancer Research, 59, 35-43.
[10] Kikuchi, S., Yamada, D., Fukami, T., Maruyama, T., Ito, A., Asamura, H., Matsuno, Y., Onizuka, M. and Murakami. Y. (2006) Hypermethylation of the TSLC1/IGSF4 Promoter Is Associated with Tobacco Smoking and a Poor Prognosis in Primary Non-Small Cell Lung Cancer. Cancer, 106, 1751-1758.
[11] Kikuchi, S., Yamada, D., Fukami, T., Masuda, M., Sakurai-Yageta, M., Williams, Y.N., Maruyama, T., Asamura, H., Matsuno, Y., Onizuka, M. and Murakami, Y. (2005) Promoter Methylation of the DAL-1/4.1B Predicts Poor Prognosis in Non-Small Cell Lung Cancer. Clinical Cancer Research, 11, 2954-2961.
[12] Murakami, Y. (2005) Involvement of a Cell Adhesion Molecule, TSLC1/IGSF4, in Human Oncogenesis. Cancer Science, 96, 543-552.
[13] Sun, C.X., Robb, V.A. and Gutmann, D.H. (2002) Protein 4.1 Tumor Suppressors: Getting a FERM Grip on Growth Regulation. Journal of Cell Science, 115, 3991-4000.
[14] Yamada, D., Kikuchi, S., Williams, Y.N., Sakurai-Yageta, M., Masuda, M., Maruyama, T., Tomita, K., Gutmann, D.H., Kakizoe, T., Kitamura, T., Kanai, Y. and Murakami, Y. (2006) Promoter Hypermethylation of the Potential Tumor Suppressor DAL-1/4.1B Gene in Renal Clear Cell Carcinoma. International Journal of Cancer, 118, 916-923.
[15] Masuda, M., Kikuchi, S., Maruyama, T., Sakurai-Yageta, M., Williams, Y.N., Ghosh, H.P. and Murakami, Y. (2005) Tumor Suppressor in Lung Cancer (TSLC)1 Suppresses Epithelial Cell Scattering and Tubulogenesis. Journal of Biological Chemistry, 280, 42164-42171.
[16] Sakurai-Yageta, M., Masuda, M., Tsuboi, Y., Ito, A. and Murakami, Y. (2009) Tumor Suppressor CADM1 Is Involved in Epithelial Cell Structure. Biochemical and Biophysical Research Communications, 390, 977-982.
[17] Ishimura, M., Sakurai-Yageta, M., Maruyama, T., Ando, T., Fukayama, M., Goto, A. and Murakami, Y. (2012) Involvement of miR-214 and miR-375 in Malignant Features of Non-Small-Cell Lung Cancer by Down-Regulating CADM1. Journal of Cancer Therapy, 3, 379-387.
[18] Shingai, T., Ikeda, W., Kakunaga, S., Morimoto, K., Takekuni, K., Takai, Y., et al. (2003) Implications of Nectin-Like Molecule-2/IGSF4/RA175/SgIGSF/TSLC1/SynCAM1 in Cell-Cell Adhesion and Transmembrane Protein Localization in Epithelial Cells. Journal of Biological Chemistry, 278, 35421-35427.
[19] Sweet-Cordero, A., Tseng, G.C., You, H., Douglass, M., Huey, B., Albertson, D. and Jacks, T. (2006) Comparison of Gene Expression and DNA Copy Number Changes in a Murine Model of Lung Cancer. Genes, Chromosomes and Cancer, 45, 338-348.
[20] Ebihara, Y., Iwai, M., Akashi, K., Ito, T., Omura, G., Saito, Y., Yoshida, M., Ando, M., Asakage, T., Yamasoba, T. and Murakami, Y. (2014) High Incidence of Null-Type Mutations of the TP53 Gene in Japanese Patients with Head and Neck Squamous Cell Carcinoma. Journal of Cancer Therapy, 5, 664-671.
[21] Burbee, D.G., Forgacs, E., Z?chbauer-Müller, S., Shivakumar, L., Fong, K., Gao, B., Randle, D., Kondo, M., Virmani, A., Bader, S., Sekido, Y., Latif, F., Milchgrub, S., Toyooka, S., Gazdar, A.F., Lerman, M.I., Zabarovsky, E., White, M. and Minna, J.D. (2001) Epigenetic Inactivation of RASSF1A in Lung and Breast Cancers and Malignant Phenotype Suppression. Journal of the National Cancer Institute, 93, 691-699.
[22] Herman, J.G., Graff, J.R., My?h?nen, S., Nelkin, B.D. and Baylin, S.B. (1996) Methylation-Specific PCR: A Novel PCR Assay for Methylation Status of CpG Islands. Proceedings of the National Academy of Sciences of the United States of America, 93, 9821-9826.
[23] House, M.G., Guo, M., Iacobuzio-Donahue, C. and Herman, J.G. (2003) Molecular Progression of Promoter Methylation in Intraductal Papillary Mucinous Neoplasms (IPMN) of the Pancreas. Carcinogenesis, 24, 193-198.
[24] Yanagawa, N., Tamura, G., Oizumi, H., Takahashi, N., Shimazaki, Y. and Motoyama, T. (2003) Promoter Hypermethylation of Tumor Suppressor and Tumor-Related Genes in Non-Small Cell Lung Cancers. Cancer Science, 94, 589-592.
[25] Guo, M., House, M.G., Akiyama, Y., Qi, Y., Capagna, D., Harmon, J., Baylin, S.B., Brock, M.V. and Herman, J.G. (2006) Hypermethylation of the GATA Gene Family in Esophageal Cancer. International Journal of Cancer, 119, 2078-2083.
[26] Suda, K., Mizuuchi, H., Maehara, Y. and Mitsudomi, T. (2012) Acquired Resistance Mechanisms to Tyrosine Kinase Inhibitors in Lung Cancer with Activating Epidermal Growth Factor Receptor Mutation-Diversity, Ductility, and Destiny. Cancer and Metastasis Reviews, 31, 807-814.
[27] Zhang, L.Q., Yin, R., Li, M., Yang, X., Jiang, F. and Xu, L. (2013) The Prognostic Value of Epigenetic Silencing of p16 Gene in NSCLC Patients: A Systematic Review and Meta-Analysis. PLoS ONE, 8, e54970.
[28] Murakami, S., Sakurai-Yageta, M., Maruyama, T. and Murakami, Y. (2014) Trans-Homophilic Interaction of CADM1 Activates PI3K by Forming a Complex with MAGuK-Family Proteins MPP3 and Dlg. PLoS ONE, 9, e82894.
[29] Toyota, M., Ahuja, N., Ohe-Toyota, M., Herman, J.G., Baylin, S.B. and Issa, J.P. (1999) CpG Island Methylator Phenotype in Colorectal Cancer. Proceedings of the National Academy of Sciences of the United States of America, 96, 8681-8686.
[30] Meng, D., Yuan, M., Li, X., Chen, L., Yang, J., Zhao, X., Ma, W. and Xin, J. (2013) Prognostic Value of K-RAS Mutations in Patients with Non-Small Cell Lung Cancer: A Systematic Review with Meta-Analysis. Lung Cancer, 81, 1-10.
[31] Jackson, E.L., Willis, N., Mercer, K., Bronson, R.T., Crowley, D., Montoya, R., Jacks, T. and Tuveson, D.A. (2001) Analysis of Lung Tumor Initiation and Progression Using Conditional Expression of Oncogenic K-Ras. Genes & Development, 15, 3243-3248.
[32] DuPage, M., Dooley, A.L. and Jacks, T. (2009) Conditional Mouse Lung Cancer Models Using Adenoviral or Lentiviral Delivery of Cre Recombinase. Nature Protocols, 4, 1064-1072.

comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.