JCT  Vol.6 No.8 , August 2015
No Association between p53 Immunohistochemical Staining and RASSF1 or DAPK1 Hypermethylation in Non-Small Cell Lung Cancer
Abstract: p53 mutations have been linked with shortened survival rates in non-small cell lung cancer (NSCLC). Hypermethylation of RASSF1 and DAPK1 genes, which are downstream targets of p53, has also been linked to a poor prognosis in lung cancer patients. We investigated whether p53 mutations, assessed as p53 stabilization by immunohistochemistry (IHC), were independent of DAPK1 and RASSF1 promoter hypermethylation. We examined 103 resected NSCLC tumors for which we had p53 IHC and RASSF1 and DAPK1 methylation data. p53 protein expression was determined by IHC and graded using a semi-quantitative scoring method. DAPK1 and RASSF1 methylations were determined on tumor blocks by MethyLight real-time PCR assays represented as the percent of methylated reference DNA (PMR). Our primary results found no evidence for an association between the p53 IHC score and RASSF1 and DAPK1 PMR values, P = 0.46 and P = 0.68, respectively.
Cite this paper: Colombara, D. , Eidsmoe, D. , Stern, J. , Feng, Q. , Vesselle, H. and Hawes, S. (2015) No Association between p53 Immunohistochemical Staining and RASSF1 or DAPK1 Hypermethylation in Non-Small Cell Lung Cancer. Journal of Cancer Therapy, 6, 631-637. doi: 10.4236/jct.2015.68069.

[1]   Tian, Y., Hou, Y., Zhou, X., et al. (2011) Tumor Suppressor RASSF1A Promoter: p53 Binding and Methylation. PLoS ONE, 6, e17017.

[2]   Zhang, H., He, J., Li, J., et al. (2013) Methylation of RASSF1A Gene Promoter Is Regulated by p53 and DAXX. FASEB Journal, 27, 232-242.

[3]   Murakami, I., Hiyama, K., Ishioka, S., et al. (2000) p53 Gene Mutations Are Associated with Shortened Survival in Patients with Advanced Non-Small Cell Lung Cancer: An Analysis of Medically Managed Patients. Clinical Cancer Research, 6, 526-530.

[4]   Feng, Q., Hawes, S.E., Stern, J.E., et al. (2008) DNA Methylation in Tumor and Matched Normal Tissues from Non-Small Cell Lung Cancer Patients. Cancer Epidemiology, Biomarkers Prevention, 17, 645-654.

[5]   De Fraipont, F., Levallet, G., Creveuil, C., et al. (2012) An Apoptosis Methylation Prognostic Signature for Early Lung Cancer in the IFCT-0002 Trial. Clinical Cancer Research, 18, 2976-2986.

[6]   Tang, X., Khuri, F.R., Lee, J.J., et al. (2000) Hypermethylation of the Death-Associated Protein (DAP) Kinase Promoter and Aggressiveness in Stage I Non-Small-Cell Lung Cancer. Journal of the National Cancer Institute, 92, 1511-1516.

[7]   Martoriati, A., Doumont, G., Alcalay, M., et al. (2005) dapk1, Encoding an Activator of a p19ARF-p53-Mediated Apoptotic Checkpoint, Is a Transcription Target of p53. Oncogene, 24, 1461-1466.

[8]   Vesselle, H., Freeman, J.D., Wiens, L., et al. (2007) Fluorodeoxyglucose Uptake of Primary Non-Small Cell Lung Cancer at Positron Emission Tomography: New Contrary Data on Prognostic Role. Clinical Cancer Research, 13, 3255-3263.

[9]   Hawes, S.E., Stern, J.E., Feng, Q., et al. (2010) DNA Hypermethylation of Tumors from Non-Small Cell Lung Cancer (NSCLC) Patients Is Associated with Gender and Histologic Type. Lung Cancer, 69, 172-179.

[10]   Kandioler-Eckersberger, D., Kappel, S., Mittlbock, M., et al. (1999) The TP53 Genotype but Not Immunohistochemical Result Is Predictive of Response to Cisplatin-Based Neoadjuvant Therapy in Stage III Non-Small Cell Lung Cancer. Journal of Thoracic and Cardiovascular Surgery, 117, 744-750.

[11]   Robles, A.I. and Harris, C.C. (2010) Clinical Outcomes and Correlates of TP53 Mutations and Cancer. Cold Spring Harbor Perspectives in Biology, 2, Article ID: a001016.

[12]   Brockenbrough, J.S., Morihara, J.K., Hawes, S.E., Stern, J.E., Rasey, J.S., Wiens, L.W., et al. (2009) Thymidine Kinase 1 and Thymidine Phosphorylase Expression in Non-Small-Cell Lung Carcinoma in Relation to Angiogenesis and Proliferation. Journal of Histochemistry and Cytochemistry, 57, 1087-1097.

[13]   Weisenberger, D.J., Campan, M., Long, T.I., Kim, M., Woods, C., Fiala, E., et al. (2005) Analysis of Repetitive Element DNA Methylation by MethyLight. Nucleic Acids Research, 33, 6823-6836.

[14]   Eads, C.A., Lord, R.V., Wickramasinghe, K., Long, T.I., Kurumboor, S.K., Bernstein, L., et al. (2001) Epigenetic Patterns in the Progression of Esophageal Adenocarcinoma. Cancer Research, 61, 3410-3418.

[15]   Ogino, S., Kawasaki, T., Brahmandam, M., Cantor, M., Kirkner, G.J., Spiegelman, D., et al. (2006) Precision and Performance Characteristics of Bisulfite Conversion and Real-Time PCR (MethyLight) for Quantitative DNA Methylation Analysis. The Journal of Molecular Diagnostics, 8, 209-217.

[16]   Yemelyanova, A., Vang, R., Kshirsagar, M., Lu, D., Marks, M.A., Shih, I.M. and Kurman, R.J. (2011) Immunohistochemical Staining Patterns of P53 Can Serve as a Surrogate Marker for TP53 Mutations in Ovarian Carcinoma: An Immunohistochemical and Nucleotide Sequencing Analysis. Modern Pathology, 24, 1248-1253.

[17]   Liu, Y., Gao, W., Siegfried, J.M., Weissfeld, J.L., Luketich, J.D. and Keohavong, P. (2007) Promoter Methylation of RASSF1A and DAPK and Mutations of K-Ras, P53, and EGFR in Lung Tumors from Smokers and Never-Smokers. BMC Cancer, 7, 74.

[18]   Lee, S.M., Lee, W.K., Kim, D.S. and Park, J.Y. (2012) Quantitative Promoter Hypermethylation Analysis of RASSF1A in Lung Cancer: Comparison with Methylation-Specific PCR Technique and Clinical Significance. Molecular Medicine Reports, 5, 239-244.

[19]   Burbee, D.G., Forgacs, E., Zochbauer-Müller, S., Shivakumar, L., Fong, K., Gao, B., et al. (2001) Epigenetic Inactivation of RASSF1A in Lung and Breast Cancers and Malignant Phenotype Suppression. Journal of the National Cancer Institute, 93, 691-699.

[20]   Kim, D.H., Nelson, H.H., Wiencke, J.K., Christiani, D.C., Wain, J.C., Mark, E.J. and Kelsey, K.T. (2001) Promoter Methylation of DAP-Kinase: Association with Advanced Stage in Non-Small Cell Lung Cancer. Oncogene, 20, 1765-1770.