DNA methylation of the p14ARF, RASSF1A and APC1A genes as an independent prognostic factor in colorectal cancer patients

Nilsson,Torbjörn  K.; Löf-Öhlin,Zarah  M.; Sun,Xiao-Feng

doi:10.3892/ijo.2012.1682

DNA methylation of the p14ARF, RASSF1A and APC1A genes as an independent prognostic factor in colorectal cancer patients

Authors:
- Torbjörn K. Nilsson
- Zarah M. Löf-Öhlin
- Xiao-Feng Sun
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Affiliations: Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden, Department of Oncology, Institute of Clinical and Experimental Medicine, Faculty of Health Sciences, University of Linköping, Linköping, Sweden
Published online on: October 30, 2012 https://doi.org/10.3892/ijo.2012.1682
Pages: 127-133
Copyright: © Nilsson et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

We quantitated the methylated fraction of CpG sites in the promoter regions of O6-MGMT, p14ARF, p16INK4a, RASSF1A and APC1A in tumor tissue from patients with colorectal cancer (CRC) in order to determine if promoter hypermethylation of any of these genes predicts survival. DNA was isolated from 111 primary CRC and 46 matched normal colorectal mucosa samples from the same patients, obtained at primary surgery and DNA methylation was examined by Pyrosequencing®. Follow-up time was up to 20 years. Patients showed partial promoter methylation in the following frequencies: O6-MGMT, 34%; p14ARF, 29%; p16INK4a, 28%; RASSF1A, 14%; and APC1A, 27%. Normal mucosa was always unmethylated. CRC patients with methylated p14ARF gene promoter had significantly worse prognosis (p=0.036), whereas those with methylated O6-MGMT had significantly better prognosis through the first 60 months post-treatment (RR 0.36; p=0.023). Methylation of one or more of the genes from the set p14ARF, RASSF1A and APC1A, was significantly (p=0.021) associated with worse prognosis even adjusting for tumor stage and differentiation (RR 2.2, p=0.037). Thus, DNA methylation of the p14ARF, RASSF1A and APC1A genes, diagnosed by Pyrosequencing, defines a poor prognosis subset of CRC patients independently of both tumor stage and differentiation. O6-MGMT methylation may play a protective role.

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1.	Das PM and Singal R: DNA methylation and cancer. J Clin Oncol. 22:4632–4642. 2004. View Article : Google Scholar : PubMed/NCBI
2.	Esteller M, Corn PG, Baylin SB and Herman JG: A gene hyper-methylation profile of human cancer. Cancer Res. 61:3225–3229. 2001.PubMed/NCBI
3.	van Engeland M, Roemen GM, Brink M, et al: K-ras mutations and RASSF1A promoter methylation in colorectal cancer. Oncogene. 21:3792–3795. 2002.
4.	Preuss I, Eberhagen I, Haas S, Eibl RH, Kaufmann M, von Minckwitz G and Kaina B: O⁶-methylguanine-DNA methyltransferase activity in breast and brain tumors. Int J Cancer. 61:321–326. 1995.
5.	Newcomb EW, Alonso M, Sung T and Miller DC: Incidence of p14^ARF gene deletion in high-grade adult and pediatric astrocytomas. Hum Pathol. 31:115–119. 2000.
6.	Serrano M, Hannon GJ and Beach D: A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature. 366:704–707. 1993. View Article : Google Scholar : PubMed/NCBI
7.	Song MS, Song SJ, Ayad NG, et al: The tumor suppressor RASSF1A regulates mitosis by inhibiting the APC-Cdc20 complex. Nat Cell Biol. 6:129–137. 2006. View Article : Google Scholar : PubMed/NCBI
8.	Rubinfeld B, Souza B, Albert I, et al: Association of the APC gene product with beta-catenin. Science. 262:1731–1734. 1993. View Article : Google Scholar : PubMed/NCBI
9.	Su LK, Vogelstein B and Kinzler KW: Association of the APC tumor suppressor protein with catenins. Science. 262:1734–1737. 1993. View Article : Google Scholar : PubMed/NCBI
10.	van den Donk M, Pellis L, Crott JW, et al: Folic acid and vitamin B-12 supplementation does not favorably influence uracil incorporation and promoter methylation in rectal mucosa DNA of subjects with previous colorectal adenomas. J Nutr. 137:2114–2120. 2007.
11.	van Engeland M, Weijenberg MP, Roemen GM, et al: Effects of dietary folate and alcohol intake on promoter methylation in sporadic colorectal cancer: the Netherlands cohort study on diet and cancer. Cancer Res. 63:3133–3137. 2003.PubMed/NCBI
12.	van den Bent MJ, Dubbink HJ, Sanson M, et al: MGMT promoter methylation is prognostic but not predictive for outcome to adjuvant PCV chemotherapy in anaplastic oligodendroglial tumors: a report from EORTC Brain Tumor Group Study 26951. J Clin Oncol. 27:5881–5886. 2009.PubMed/NCBI
13.	Löf-Öhlin ZM and Nilsson TK: Pyrosequencing assays to study promoter CpG site methylation of the O⁶-MGMT, hMLH1, p14^ARF, p16^INK4a, RASSF1A and APC1A genes. Oncol Rep. 21:721–729. 2009.PubMed/NCBI
14.	Aoyagi H, Iida S, Uetake H, et al: Effect of classification based on combination of mutation and methylation in colorectal cancer prognosis. Oncol Rep. 25:789–794. 2011.PubMed/NCBI
15.	Esteller M, Tortola S, Toyota M, Capella G, Peinado MA, Baylin SB and Herman JG: Hypermethylation-associated inactivation of p14(ARF) is independent of p16(INK4a) methylation and p53 mutational status. Cancer Res. 60:129–133. 2000.PubMed/NCBI
16.	Kamiyama H, Noda H, Takata O, Suzuki K, Kawamura Y and Konishi F: Promoter hypermethylation of tumor-related genes in peritoneal lavage and the prognosis of patients with colorectal cancer. J Surg Oncol. 100:69–74. 2009. View Article : Google Scholar : PubMed/NCBI
17.	Chen SP, Chiu SC, Wu CC, et al: The association of methylation in the promoter of APC and MGMT and the prognosis of Taiwanese CRC patients. Genet Test Mol Biomarkers. 13:67–71. 2009. View Article : Google Scholar : PubMed/NCBI
18.	Noda H, Mashima R, Kamiyama H, Okada S, Kawamura YJ and Konishi F: Promoter hypermethylation of tumor-related genes in sporadic colorectal cancer in young patients. J Exp Clin Cancer Res. 26:521–526. 2007.PubMed/NCBI
19.	Krtolica K, Krajnovic M, Usaj-Knezevic S, Babic D, Jovanovic D and Dimitrijevic B: Comethylation of p16 and MGMT genes in colorectal carcinoma: correlation with clinicopathological features and prognostic value. World J Gastroenterol. 13:1187–1194. 2007. View Article : Google Scholar : PubMed/NCBI
20.	Kohonen-Corish MR, Daniel JJ, Chan C, et al: Low microsatellite instability is associated with poor prognosis in stage C colon cancer. J Clin Oncol. 23:2318–2324. 2005. View Article : Google Scholar : PubMed/NCBI
21.	Nakayama G, Hibi K, Kodera Y, Koike M, Fujiwara M and Nakao A: P16 methylation in serum as a potential marker for the malignancy of colorectal carcinoma. Anticancer Res. 27:3367–3370. 2007.PubMed/NCBI
22.	Lee M, Han WS, Kim OK, Sung SH, Cho MS, Lee SN and Koo H: Prognostic value of p16^INK4a and p14^ARF gene hypermethylation in human colon cancer. Pathol Res Pract. 202:415–424. 2006.
23.	Sanz-Casla MT, Maestro ML, Vidaurreta M, Maestro C, Arroyo M and Cerdan J: p16 Gene methylation in colorectal tumors: correlation with clinicopathological features and prognostic value. Dig Dis. 23:151–155. 2005. View Article : Google Scholar : PubMed/NCBI
24.	Zou HZ, Yu BM, Wang ZW, et al: Detection of aberrant p16 methylation in the serum of colorectal cancer patients. Clin Cancer Res. 8:188–191. 2002.
25.	Dominguez G, Silva J, Garcia JM, et al: Prevalence of aberrant methylation of p14^ARF over p16^INK4a in some human primary tumors. Mutat Res. 530:9–17. 2003. View Article : Google Scholar : PubMed/NCBI
26.	Olschwang S, Hamelin R, Laurent-Puig P, et al: Alternative genetic pathways in colorectal carcinogenesis. Proc Natl Acad Sci USA. 94:12122–12127. 1997. View Article : Google Scholar : PubMed/NCBI
27.	Shen L, Catalano PJ, Benson AB III, O’Dwyer P, Hamilton SR and Issa JP: Association between DNA methylation and shortened survival in patients with advanced colorectal cancer treated with 5-fluorouracil based chemotherapy. Clin Cancer Res. 13:6093–6098. 2007. View Article : Google Scholar : PubMed/NCBI
28.	Hoffmann AC, Vallböhmer D, Prenzel K, et al: Methylated DAPK and APC promoter DNA detection in peripheral blood is a significantly associated with apparent residual tumor and outcome. J Cancer Res Clin Oncol. 135:1231–1237. 2009. View Article : Google Scholar : PubMed/NCBI
29.	Kim JC, Choi JS, Roh SA, Cho DH, Kim TW and Kim YS: Promoter methylation of specific genes is associated with the phenotype and progression of colorectal adenocarcinomas. Ann Surg Oncol. 17:1767–1776. 2010. View Article : Google Scholar : PubMed/NCBI
30.	Wettergren Y, Odin E, Nilsson S, Carlsson G and Gustavsson B: p16^INK4a gene promoter hypermethylation in mucosa as a prognostic factor for patients with colorectal cancer. Mol Med. 14:412–421. 2008. View Article : Google Scholar
31.	Tommasi S, Pinto R, Petriella D, et al: Oncosuppressor methylation: a possible key role in colon metastatic progression. J Cell Physiol. 226:1934–1939. 2011. View Article : Google Scholar : PubMed/NCBI
32.	Weller M, Felsberg J, Hartmann C, et al: Molecular predictors of progression-free and overall survival in patients with newly diagnosed glioblastoma: a prospective translational study of the German Glioma Network. J Clin Oncol. 27:5743–5750. 2009. View Article : Google Scholar
33.	Hegi ME, Diserens AC, Gorlia T, et al: MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med. 352:997–1003. 2005. View Article : Google Scholar : PubMed/NCBI
34.	Esteller M, Garcia-Foncillas J, Andion E, et al: Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. N Engl J Med. 343:1350–1354. 2000. View Article : Google Scholar
35.	Jaeckle KA, Eyre HJ, Townsend JJ, et al: Correlation of tumor O⁶ methylguanine-DNA methyltransferase levels with survival of malignant astrocytoma patients treated with bis-chloroethylnitrosourea: a Southwest Oncology Group study. J Clin Oncol. 16:3310–3315. 1998.
36.	Hegi ME, Diserens AC, Godard S, et al: Clinical trial substantiates the predictive value of O-6-methylguanine-DNA methyltransferase promoter methylation in glioblastoma patients treated with temozolomide. Clin Cancer Res. 10:1871–1874. 2004. View Article : Google Scholar : PubMed/NCBI
37.	Stupp R and Hegi ME: Methylguanine methyltransferase testing in glioblastoma: when and how? J Clin Oncol. 25:1459–1460. 2007. View Article : Google Scholar : PubMed/NCBI
38.	Shima K, Morikawa T, Baba Y, et al: MGMT promoter methylation, loss of expression and prognosis in 855 colorectal cancers. Cancer Causes Control. 22:301–309. 2011. View Article : Google Scholar : PubMed/NCBI
39.	Löf-Öhlin ZM, Sorbe B, Wingren S and Nilsson TK: Hypermethylation of promoter regions of the APC1A and p16^INK4a genes in relation to prognosis and tumor characteristics in cervical cancer patients. Int J Oncol. 39:683–688. 2011.
40.	Carmona FJ and Esteller M: Moving closer to a prognostic DNA methylation signature in colon cancer. Clin Cancer Res. 17:1215–1217. 2011. View Article : Google Scholar : PubMed/NCBI
41.	Yi JM, Dhir M, Van Neste L, et al: Genomic and epigenomic integration identifies a prognostic signature in colon cancer. Clin Cancer Res. 17:1535–1545. 2011. View Article : Google Scholar : PubMed/NCBI
42.	Cui T, Chen Y, Yang L, Knösel T, Zöller K, Huber O and Petersen I: DSC3 expression is regulated by p53, and methylation of DSC3 DNA is a prognostic marker in human colorectal cancer. Br J Cancer. 104:1013–1019. 2011. View Article : Google Scholar : PubMed/NCBI
43.	Tong JD, Jiao NL, Wang YX, Zhang YW and Han F: Downregulation of fibulin-3 gene by promoter methylation in colorectal cancer predicts adverse prognosis. Neoplasma. 58:441–448. 2011. View Article : Google Scholar : PubMed/NCBI
44.	van Engeland M, Derks S, Smits KM, Meijer GA and Herman JG: Colorectal cancer epigenetics: complex simplicity. J Clin Oncol. 29:1382–1391. 2011.PubMed/NCBI
45.	Jover R, Nguyen TP, Pérez-Carbonell L, et al: 5-Fluorouracil adjuvant chemotherapy does not increase survival in patients with CpG island methylator phenotype colorectal cancer. Gastroenterology. 140:1174–1181. 2011. View Article : Google Scholar : PubMed/NCBI