Methylation status of p16 and p14 genes in locally advanced rectal cancer: potential clinical implication
Keywords:rectal cancer, p16, p14, DNA methylation, biomarkers, VEGF, EGFR
- Patients’ response to standard treatment of locally advanced rectal cancer is variable. Many tested molecular parameters have failed to become reliable predictive biomarkers. Promotor hypermethylation of p16 and p14 genes has been frequently reported as an early event in colorectal cancer, but its clinical relevance in rectal cancer, as a distinct entity, has not been established.
- Combined analysis of p16 and p14 methylation and VEGF expression status could potentially predict a more aggressive phenotype of locally advanced rectal cancer.
- Aberrant p16 or p14 methylation, together with high vascular endothelial growth factor (VEGF) expression, is significantly associated with a worse treatment response.
Abstract: Methylation of p16 and p14 genes is a common event in colorectal cancers; however, their exact role in the prediction of patients’ outcome is unclear. We conducted this retrospective study to evaluate their potential predictive and/or prognostic roles. Methylation-specific PCR was used to examine the methylation status of p16 and p14 in pretherapeutic and preoperative biopsy specimens of 60 patients with locally advanced rectal cancer. The methylation status of the examined genes did not affect the response to preoperative chemoradiotherapy (CRT), recurrence rate and overall survival. However, patients with a simultaneous presence of either p16 or p14 methylation and high vascular endothelial growth factor (VEGF) expression showed a significantly worse response to CRT (p=0.005 and p=0.038, respectively). Moreover, patients with both p16 methylation and high VEGF expression had significantly shorter overall survival (p=0.010), while no such association was found in patients with p14 methylation and high VEGF expression. On the other hand, a subgroup of patients with p16 methylation and low VEGF and high epidermal growth factor receptor (EGFR) expression showed a significantly better response to CRT (p=0.024). The obtained results point to the importance of p16 and p14 methylation analyses in combination with VEGF and EGFR expression, aimed at better predicting treatment response and patient outcome.
Received: March 16, 2018; Revised: June 13, 2018; Accepted: June 13, 2018; Published online: June 19, 2018
How to cite this article: Kožik B, Kokanov N, Knežević-Ušaj S, Nikolić I, Davidović R, Jovanović-Ćupić S, Krajnović M. Methylation status of p16 and p14 genes in locally advanced rectal cancer: Potential clinical implication. Arch Biol Sci. 2018;70(4):681-90.
Schmoll HJ, Cutsem E Van, Stein A, Valentini V, Glimelius B, Haustermans K, Nordlinger B, van de Velde CJ, Balmana J, Regula J, Nagtegaal ID, Beets-Tan RG, Arnold D, Ciardiello F, Hoff P, Kerr D, Köhne CH, Labianca R, Price T, Scheithauer W, Sobrero A, Tabernero J, Aderka D, Barroso S, Bodoky G, Douillard JY, El Ghazaly H, Gallardo J, Garin A, Glynne-Jones R, Jordan K, Meshcheryakov A, Papamichail D, Pfeiffer P, Souglakos I, Turhal S, Cervantes A. ESMO Consensus Guidelines for management of patients with colon and rectal cancer. A personalized approach to clinical decision making. Ann Oncol. 2012;23:2479-516.
Glimelius B. Neo-adjuvant radiotherapy in rectal cancer. World J Gastroenterol. 2013;19(46):8489-501.
Martin ST, Heneghan HM, Winter DC. Systematic review and meta-analysis of outcomes following pathological complete response to neoadjuvant chemoradiotherapy for rectal cancer. Br J Surg. 2012;99:918-28.
Milgrom SA, García-Aguilar J. Molecular biomarkers as predictors of response to neoadjuvant chemoradiation therapy in rectal cancer. Semin Colon Rectal Surg. 2013;119-24.
Zeestraten EC, Kuppen PJ, van de Velde CJ, Marijnen CA. Prediction in rectal cancer. Semin Radiat Oncol. 2012;22(2):175-83.
Sun W. Angiogenesis in metastatic colorectal cancer and the benefits of targeted therapy. J Hematol Oncol. 2012;5:1-9.
Coppedè F. Epigenetic biomarkers of colorectal cancer: Focus on DNA methylation. Cancer Lett. 2014;342:238-47.
Sakai E, Nakajima A, Kaneda A. Accumulation of aberrant DNA methylation during colorectal cancer development. World J Gastroenterol. 2014;20:978-87.
Ng J, Yu J. Promoter Hypermethylation of Tumour Suppressor Genes as Potential Biomarkers in Colorectal Cancer. Int J Mol Sci. 2015;16:2472-96.
Draht MX, Riedl RR, Niessen H, Carvalho B, Meijer GA, Herman JG, van Engeland M, Melotte V, Smits KM. Promoter CpG island methylation markers in colorectal cancer: the road ahead. Epigenomics. 2012;4(2):179-94.
Sharpless N. INK4a/ARF: a multifunctional tumor suppressor locus. Mutat Res. 2005;576(1-2):22-38.
Sharpless N, DePinho R. The INK4A/ARF locus and its two gene products. Curr Opin Genet Dev. 1999;9:22-30.
Lu Y, Zhang X, Zhang J. Inhibition of Breast Tumor Cell Growth by Ectopic Expression of p16/INK4A Via Combined Effects of Cell Cycle Arrest, Senescence and Apoptotic Induction, and Angiogenesis Inhibition. J Cancer. 2012;3:333-44.
Kawagishi H, Nakamura H, Maruyama M, Mizutani S, Sugimoto K, Takagi M, Sugimoto M. ARF suppresses tumor angiogenesis through translational control of VEGFA mRNA. Cancer Res. 2010;70:4749-58.
Veganzones-de-Castro S, Rafael-Fernández S, Vidaurreta-Lázaro M, Orden V, Mediero-Valeros B, Fernández C, Maestro-de las Casas ML. p16 gene methylation in colorectal cancer patients with long-term follow-up. Rev Esp Enferm Dig. 2012;104:111-7.
Nyiraneza C, Sempoux C, Detry R, Kartheuser A, Dahan K. Hypermethylation of the 5’ CpG island of the p14ARF flanking exon 1β in human colorectal cancer displaying a restricted pattern of p53 overexpression concomitant with increased MDM2 expression. Clin Epigenetics. 2012;4:9.
Sambrook J, Fritsch EF, Maniatis T. Analysis and cloning of eukariotic genomic DNA. In: Ford N, Nolan C, Ferguson M, editors. Molecular Cloning, A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press; 1989. p.16-9.
Herman J, Graff J, Myöhänen S, Nelkin B, Baylin S. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A. 1996;93:9821-6.
Esteller M, Tortola S, Toyota M, Capella G, Peinado MA, Baylin SB, Herman JG. Hypermethylation-associated inactivation of p14(ARF) is independent of p16(INK4a) methylation and p53 mutational status. Cancer Res. 2000;60:129-33.
Krajnović M, Marković B, Knežević-Ušaj S, Nikolić I, Stanojević M, Nikolić V, Šiljić M, Jovanović Ćupić S, Dimitrijević B. Locally advanced rectal cancers with simultaneous occurrence of KRAS mutation and high VEGF expression show invasive characteristics. Pathol Res Pract. 2016;212:598-603.
Azad N, Zahnow CA, Rudin CM, Baylin SB. The future of epigenetic therapy in solid tumours-lessons from the past. Nat Rev Clin Oncol. 2013;10:256-66.
Xing X, Cai W, Shi H, Wang Y, Li M, Jiao J, Chen M. The prognostic value of CDKN2A hypermethylation in colorectal cancer: a meta-analysis. Br J Cancer. 2013;108:2542-8.
Hibi K, Nakayama H, Koike M, Kasai Y, Ito K, Akiyama S, Nakao A. Colorectal Cancers with both p16 and p14 Methylation Show Invasive Characteristics. Jpn J Cancer Res. 2002;93:883-7.
Kang MY, Lee BB, Ji YI, Jung EH, Chun HK, Song SY, Park SE, Park J, Kim DH. Association of interindividual differences in p14ARF promoter methylation with single nucleotide polymorphism in primary colorectal cancer. Cancer. 2008;112(8):1699-707.
Lee M, Sup Han W, Kyoung Kim O, Hee Sung S, Sun Cho M, Lee SN, Koo H. Prognostic value of p16INK4a and p14ARF gene hypermethylation in human colon cancer. Pathol Res Pract. 2006;202:415-24.
Jiang W, Wang PG, Zhan Y, Zhang D. Prognostic value of p16 promoter hypermethylation in colorectal cancer: a meta-analysis. Cancer Invest. 2014;32(2):43-52.
Kim JC, Choi JS, Roh SA, Cho DH, Kim TW, Kim YS. Promoter methylation of specific genes is associated with the phenotype and progression of colorectal adenocarcinomas. Ann Surg Oncol. 2010;17:1767-76.
Zhou Z, Zhang H, Lai J, Diao D, Li W, Dang C, Song Y. Relationships between p14ARF gene methylation and clinicopathological features of colorectal cancer: a meta-analysis. PLoS One. 2016;11(3):e0152050.
Nilsson TKK, Löf-Öhlin ZM, Sun X-FF. DNA methylation of the p14ARF, RASSF1A and APC1A genes as an independent prognostic factor in colorectal cancer patients. Int J Oncol. 2013;42:127-33.
Hibi K, Nakao A. Lymph node metastasis is infrequent in patients with highly-methylated colorectal cancer. Anticancer Res. 2006;26:55-8.
Lee MS, Menter DG, Kopetz S. Right Versus Left Colon Cancer Biology: Integrating the Consensus Molecular Subtypes. J Natl Compr Canc Netw. 2017;15(3):411-9.
Kohonen-Corish MR, Tseung J, Chan C, Currey N, Dent OF, Clarke S, Bokey L, Chapuis PH. KRAS mutations and CDKN2A promoter methylation show an interactive adverse effect on survival and predict recurrence of rectal cancer. Int J Cancer. 2014;134:2820-8.
Giatromanolaki A, Sivridis E, Koukourakis MI. Angiogenesis in colorectal cancer: prognostic and therapeutic implications. Am J Clin Oncol. 2006;29:408-17.
Giralt J, Navalpotro B, Hermosilla E, de Torres I, Espin E, Reyes V, Cerezo L, de las Heras M, Ramon y Cajal S, Armengol M, Benavente S. Prognostic significance of vascular endothelial growth factor and cyclooxygenase-2 in patients with rectal cancer treated with preoperative radiotherapy. Oncology. 2007;71:312-9.
Figueras A, Arbos MA, Quiles MT, Vinals F, Germa JR, Capella G. The impact of KRAS mutations on VEGF-A production and tumour vascular network. BMC Cancer. 2013;13:125.
Amelio I, Melino G. The p53 family and the hypoxia-inducible factors (HIFs): determinants of cancer progression. Trends Biochem Sci. 2015;40(8):425-34.
Kapiteijn E, Liefers GJ, Los LC, Kranenbarg EK, Hermans J, Tollenaar RA, Moriya Y, van de Velde CJ, van Krieken JH. Mechanisms of oncogenesis in colon versus rectal cancer. J Pathol. 2001;195:171-8.
Takeuchi H, Ozawa S, Shih C-H, Ando N, Kitagawa Y, Ueda M, Kitajima M. Loss of p16INK4a expression is associated with vascular endothelial growth factor expression in squamous cell carcinoma of the esophagus. Int J Cancer J Int Du Cancer. 2004;109:483-90.
Kim JS, Kim JM, Li S, Yoon WH, Song KS, Kim KH, Yeo SG, Nam JS, Cho MJ. Epidermal growth factor receptor as a predictor of tumor downstaging in locally advanced rectal cancer patients treated with preoperative chemoradiotherapy. Int J Radiat Oncol Biol Phys. 2006;66:195-200.
Giralt J, de las Heras M, Cerezo L, Eraso A, Hermosilla E, Velez D, Lujan J, Espin E, Rosello J, Majó J, Benavente S, Armengol M, de Torres I; Grupo Español de Investigacion Clinica en Oncologia Radioterápica (GICOR). The expression of epidermal growth factor receptor results in a worse prognosis for patients with rectal cancer treated with preoperative radiotherapy: a multicenter, retrospective analysis. Radiother Oncol. 2005;74(2):101-8.
Li S, Kim JS, Kim JM, Cho MJ, Yoon WH, Song KS, Yeo SG, Kim JS. Epidermal growth factor receptor as a prognostic factor in locally advanced rectal-cancer patients treated with preoperative chemoradiation. Int J Radiat Oncol Biol Phys. 2006;65:705-12.
Zlobec I, Vuong T, Compton CC, Lugli A, Michel RP, Hayashi S, Jass JR. Combined analysis of VEGF and EGFR predicts complete tumour response in rectal cancer treated with preoperative radiotherapy. Br J Cancer. 2008;98:450-6.
Chakravarti A, Winter K, Wu CL, Kaufman D, Hammond E, Parliament M, Tester W, Hagan M, Grignon D, Heney N, Pollack A, Sandler H, Shipley W. Expression of the epidermal growth factor receptor and Her-2 are predictors of favorable outcome and reduced complete response rates, respectively, in patients with muscle-invading bladder cancers treated by concurent radiation and cisplatin-based chemotherapy: a report from the radiation therapy oncology group. Int J Radiation Oncology Biol Phys. 2005;62:309-17.
Roberts PJ, Der CJ. Targeting the Raf - MEK - ERK mitogen-activated protein kinase cascade for the treatment of cancer. Oncogene. 2007;26:3291-310.
Ciardiello F, Troiani T, Bianco R, Orditura M, Morgillo F, Martinelli E, Morelli MP, Cascone T, Tortora G. Interaction between the epidermal growth factor receptor (EGFR) and the vascular endothelial growth factor (VEGF) pathways: a rational approach for multi-target anticancer therapy. Ann Oncol. 2006;17:109-14.