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Published in: Pathology & Oncology Research 1/2017

01-01-2017 | Original Article

Alterations of p14 ARF , p15 INK4b , and p16 INK4a Genes in Primary Laryngeal Squamous Cell Carcinoma

Authors: Fernando López, Teresa Sampedro, José L. Llorente, Mario Hermsen, César Álvarez-Marcos

Published in: Pathology & Oncology Research | Issue 1/2017

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Abstract

The 9p21 gene cluster, harboring growth suppressive genes p14 ARF , p15 INK4b , and p16 INK4a , is one of the major aberration hotspots in head and neck cancers. We try to elucidate specific aberrations affecting this region, throughout methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay. Methylation of the gene was investigated by MS-MLPA in a well-characterized series of 27 laryngeal squamous cell carcinomas and 20 samples of healthy mucosa. Aberrant promoter hypermethylation was confirmed using and methylation-specific. All samples studied except 3 (11 %) presented losses at 9p21 segment. The most common finding was the small deletion (exon 1α) of the p16 INK4a locus (44 %). Deletion of the 9p21 gene cluster was identified in 5 cases (18 %). We only found methylation in 8 samples (30 %) for p15 IK4b -exon 1. Promoter methylation of p14 ARF , p15 IK4b and p16 INK4a was not detected in any tumor sample. Methylation-specific polymerase chain reaction confirmed the results. Our data indicate that there may be a subgroup of patients in which epigenetic regulation of 9p21 segment might have little relevance. Nevertheless, MS-MLPA could not be suitable for the study of methylation at this region and further research is required.
Literature
1.
go back to reference Worsham MJ, Chen KM, Tiwari N, Pals G, Schouten JP, Sethi S, et al. (2006) Fine-mapping loss of gene architecture at the CDKN2B (p15INK4b), CDKN2A (p14ARF, p16INK4a), and MTAP genes in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg 132:409–415CrossRefPubMed Worsham MJ, Chen KM, Tiwari N, Pals G, Schouten JP, Sethi S, et al. (2006) Fine-mapping loss of gene architecture at the CDKN2B (p15INK4b), CDKN2A (p14ARF, p16INK4a), and MTAP genes in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg 132:409–415CrossRefPubMed
2.
go back to reference Marcos CÁ, Alonso-Guervós M, Prado NR, Gimeno TS, Iglesias FD, Hermsen M, et al. (2011) Genetic model of transformation and neoplastic progression in laryngeal epithelium. Head Neck 33:216–224CrossRefPubMed Marcos CÁ, Alonso-Guervós M, Prado NR, Gimeno TS, Iglesias FD, Hermsen M, et al. (2011) Genetic model of transformation and neoplastic progression in laryngeal epithelium. Head Neck 33:216–224CrossRefPubMed
3.
go back to reference Laytragoon-Lewin N, Chen F, Castro J, Elmberger G, Rutqvist LE, Lewin F, et al. (2010) DNA content and methylation of p16, DAPK and RASSF1A gene in tumour and distant, normal mucosal tissue of head and neck squamous cell carcinoma patients. Anticancer Res 30:4643–4648PubMed Laytragoon-Lewin N, Chen F, Castro J, Elmberger G, Rutqvist LE, Lewin F, et al. (2010) DNA content and methylation of p16, DAPK and RASSF1A gene in tumour and distant, normal mucosal tissue of head and neck squamous cell carcinoma patients. Anticancer Res 30:4643–4648PubMed
4.
go back to reference Xing EP, Nie Y, Song Y, Yang GY, Cai YC, Wang LD, et al. (1999) Mechanisms of inactivation of p14ARF, p15INK4b, and p16INK4a genes in human esophageal squamous cell carcinoma. Clin Cancer Res 5:2704–2713PubMed Xing EP, Nie Y, Song Y, Yang GY, Cai YC, Wang LD, et al. (1999) Mechanisms of inactivation of p14ARF, p15INK4b, and p16INK4a genes in human esophageal squamous cell carcinoma. Clin Cancer Res 5:2704–2713PubMed
6.
go back to reference Chen K, Sawhney R, Khan M, Benninger MS, Hou Z, Sethi S, et al. (2007) Methylation of multiple genes as diagnostic and therapeutic markers in primary head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg 133:1131–1138CrossRefPubMed Chen K, Sawhney R, Khan M, Benninger MS, Hou Z, Sethi S, et al. (2007) Methylation of multiple genes as diagnostic and therapeutic markers in primary head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg 133:1131–1138CrossRefPubMed
7.
go back to reference López F, Sampedro T, Llorente JL, Domínguez F, Hermsen M, Suárez C, et al. (2014) Utility of MS-MLPA in DNA methylation profiling in primary laryngeal squamous cell carcinoma. Oral Oncol 50:291–297CrossRefPubMed López F, Sampedro T, Llorente JL, Domínguez F, Hermsen M, Suárez C, et al. (2014) Utility of MS-MLPA in DNA methylation profiling in primary laryngeal squamous cell carcinoma. Oral Oncol 50:291–297CrossRefPubMed
8.
go back to reference López F, Llorente JL, García-Inclán C, Alonso-Guervós M, Cuesta-Albalad MP, Fresno MF, et al. (2011) Genomic profiling of sinonasal squamous cell carcinoma. Head Neck 33:145–153CrossRefPubMed López F, Llorente JL, García-Inclán C, Alonso-Guervós M, Cuesta-Albalad MP, Fresno MF, et al. (2011) Genomic profiling of sinonasal squamous cell carcinoma. Head Neck 33:145–153CrossRefPubMed
9.
go back to reference Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G (2002) Relative quantification of 40 nucleic acid sequences by multiplex ligation dependent probe amplification. Nucl Acids Res 30:e57CrossRefPubMedPubMedCentral Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G (2002) Relative quantification of 40 nucleic acid sequences by multiplex ligation dependent probe amplification. Nucl Acids Res 30:e57CrossRefPubMedPubMedCentral
10.
go back to reference Nygren AO, Ameziane N, Duarte HM, Vijzelaar RN, Waisfisz Q, Hess CJ, et al. (2005) Methylation-specific MLPA (MS-MLPA): simultaneous detection of CpG methylation and copy number changes of up to 40 sequences. Nucl Acids Res 33:e128CrossRefPubMedPubMedCentral Nygren AO, Ameziane N, Duarte HM, Vijzelaar RN, Waisfisz Q, Hess CJ, et al. (2005) Methylation-specific MLPA (MS-MLPA): simultaneous detection of CpG methylation and copy number changes of up to 40 sequences. Nucl Acids Res 33:e128CrossRefPubMedPubMedCentral
11.
go back to reference Furonaka O, Takeshima Y, Awaya H, Ishida H, Kohno N, Inai K (2004) Aberrant methylation of p 14(ARF), p 15(INK4b) and p 16(INK4a) Genes and location of the primary site in pulmonary squamous cell carcinoma. Pathol Int 54:549–555CrossRefPubMed Furonaka O, Takeshima Y, Awaya H, Ishida H, Kohno N, Inai K (2004) Aberrant methylation of p 14(ARF), p 15(INK4b) and p 16(INK4a) Genes and location of the primary site in pulmonary squamous cell carcinoma. Pathol Int 54:549–555CrossRefPubMed
12.
go back to reference Geradts J, Wilson PA (1996) High frequency of aberrant p16(INK4A) expression in human breast cancer. Am J Pathol 149:15–20PubMedPubMedCentral Geradts J, Wilson PA (1996) High frequency of aberrant p16(INK4A) expression in human breast cancer. Am J Pathol 149:15–20PubMedPubMedCentral
13.
go back to reference Cabanillas R, Rodrigo JP, Ferlito A, Rinaldo A, Fresno MF, Aguilar C, et al. (2007) Is there an epidemiological link between human papillomavirus DNA and basaloid squamous cell carcinoma of the pharynx? Oral Oncol 43:327–332CrossRefPubMed Cabanillas R, Rodrigo JP, Ferlito A, Rinaldo A, Fresno MF, Aguilar C, et al. (2007) Is there an epidemiological link between human papillomavirus DNA and basaloid squamous cell carcinoma of the pharynx? Oral Oncol 43:327–332CrossRefPubMed
14.
go back to reference Danahey DG, Tobin EJ, Schuller DE, Bier-Laning CM, Weghorst CM, Lang JC (1999) p16 Mutation frequency and clinical correlation in head and neck cancer. Acta Otolaryngol 119:285–288CrossRefPubMed Danahey DG, Tobin EJ, Schuller DE, Bier-Laning CM, Weghorst CM, Lang JC (1999) p16 Mutation frequency and clinical correlation in head and neck cancer. Acta Otolaryngol 119:285–288CrossRefPubMed
15.
go back to reference Shintani S, Nakahara Y, Mihara M, Ueyama Y, Matsumura T (2001) Inactivation of the p14(ARF), p15(INK4B) And p16(INK4A) genes is a frequent event in human oral squamous cell carcinomas. Oral Oncol 37:498–504CrossRefPubMed Shintani S, Nakahara Y, Mihara M, Ueyama Y, Matsumura T (2001) Inactivation of the p14(ARF), p15(INK4B) And p16(INK4A) genes is a frequent event in human oral squamous cell carcinomas. Oral Oncol 37:498–504CrossRefPubMed
16.
go back to reference Stephen JK, Chen KM, Shah V, Havard S, Kapke A, Lu M, et al. (2010) DNA hypermethylation markers of poor outcome in laryngeal cancer. Clin Epigenetics 1:61–69CrossRefPubMedPubMedCentral Stephen JK, Chen KM, Shah V, Havard S, Kapke A, Lu M, et al. (2010) DNA hypermethylation markers of poor outcome in laryngeal cancer. Clin Epigenetics 1:61–69CrossRefPubMedPubMedCentral
17.
go back to reference Wong TS, Man MWL, Lam AKY, Wei WI, Kwong YL, Yuen APW (2003) The study of p16 and p15 gene methylation in head and neck squamous cell carcinoma and their quantitative evaluation in plasma by real-time PCR. Eur J Cancer 39:1881–1887CrossRefPubMed Wong TS, Man MWL, Lam AKY, Wei WI, Kwong YL, Yuen APW (2003) The study of p16 and p15 gene methylation in head and neck squamous cell carcinoma and their quantitative evaluation in plasma by real-time PCR. Eur J Cancer 39:1881–1887CrossRefPubMed
18.
go back to reference Pierini S, Jordanov SH, Mitkova AV, Chalakov IJ, Melnicharov MB, Kunev KV, et al. (2014) Promoter hypermethylation of CDKN2A, MGMT, MLH1 and DAPK genes in laryngeal squamous cell carcinoma and their associations with clinical profiles of the patients. Head Neck 36:1103–1108CrossRefPubMed Pierini S, Jordanov SH, Mitkova AV, Chalakov IJ, Melnicharov MB, Kunev KV, et al. (2014) Promoter hypermethylation of CDKN2A, MGMT, MLH1 and DAPK genes in laryngeal squamous cell carcinoma and their associations with clinical profiles of the patients. Head Neck 36:1103–1108CrossRefPubMed
19.
go back to reference Li X, Gao L, Li H, Gao J, Yang Y, Zhou F, et al. (2013) Human papillomavirus infection and laryngeal cancer risk: a systematic review and meta-analysis. J Infect Dis 207:479–488CrossRefPubMed Li X, Gao L, Li H, Gao J, Yang Y, Zhou F, et al. (2013) Human papillomavirus infection and laryngeal cancer risk: a systematic review and meta-analysis. J Infect Dis 207:479–488CrossRefPubMed
20.
go back to reference Rodrigo JP, Hermsen MA, Fresno MF, Brakenhoff RH, García-Velasco F, Snijders PJ, et al. (2015) Prevalence of human papillomavirus in laryngeal and hypopharyngeal squamous cell carcinomas in northern Spain. Cancer Epidemiol 39:37–41CrossRefPubMed Rodrigo JP, Hermsen MA, Fresno MF, Brakenhoff RH, García-Velasco F, Snijders PJ, et al. (2015) Prevalence of human papillomavirus in laryngeal and hypopharyngeal squamous cell carcinomas in northern Spain. Cancer Epidemiol 39:37–41CrossRefPubMed
21.
go back to reference Ohta S, Uemura H, Matsui Y, Ishiguro H, Fujinami K, Kondo K, et al. (2009) Alterations of p16 and p14ARF genes and their 9p21 locus in oral squamous cell carcinoma. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:81–91CrossRefPubMed Ohta S, Uemura H, Matsui Y, Ishiguro H, Fujinami K, Kondo K, et al. (2009) Alterations of p16 and p14ARF genes and their 9p21 locus in oral squamous cell carcinoma. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:81–91CrossRefPubMed
22.
go back to reference Ignatov A, Bischoff J, Schwarzenau C, Krebs T, Kuester D, Herrmann K, et al. (2008) p16 alterations increase the metastatic potential of endometrial carcinoma. Gynecol Oncol 111:365–371CrossRefPubMed Ignatov A, Bischoff J, Schwarzenau C, Krebs T, Kuester D, Herrmann K, et al. (2008) p16 alterations increase the metastatic potential of endometrial carcinoma. Gynecol Oncol 111:365–371CrossRefPubMed
23.
go back to reference Kobayashi N, Toyooka S, Yanai H, Soh J, Fujimoto N, Yamamoto H, et al. (2008) Frequent p16 inactivation by homozygous deletion or methylation is associated with a poor prognosis in Japanese patients with pleural mesothelioma. Lung Cancer 62:120–125CrossRefPubMed Kobayashi N, Toyooka S, Yanai H, Soh J, Fujimoto N, Yamamoto H, et al. (2008) Frequent p16 inactivation by homozygous deletion or methylation is associated with a poor prognosis in Japanese patients with pleural mesothelioma. Lung Cancer 62:120–125CrossRefPubMed
24.
go back to reference 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 U S A 93:9821–9826CrossRefPubMedPubMedCentral 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 U S A 93:9821–9826CrossRefPubMedPubMedCentral
25.
go back to reference Wojdacz TK, Dobrovic A (2007) Methylation-sensitive high resolution melting (MS-HRM): a new approach for sensitive and high-throughput assessment of methylation. Nucleic Acids Res 35(6):e41CrossRefPubMedPubMedCentral Wojdacz TK, Dobrovic A (2007) Methylation-sensitive high resolution melting (MS-HRM): a new approach for sensitive and high-throughput assessment of methylation. Nucleic Acids Res 35(6):e41CrossRefPubMedPubMedCentral
26.
go back to reference Pavicic W, Perkiö E, Kaur S, Peltomäki P (2011) Altered methylation at microRNA-associated CpG islands in hereditary and sporadic carcinomas: a methylation-specific multiplex ligation dependent probe amplification (MS-MLPA)-based approach. Mol Med 17:726–735CrossRefPubMedPubMedCentral Pavicic W, Perkiö E, Kaur S, Peltomäki P (2011) Altered methylation at microRNA-associated CpG islands in hereditary and sporadic carcinomas: a methylation-specific multiplex ligation dependent probe amplification (MS-MLPA)-based approach. Mol Med 17:726–735CrossRefPubMedPubMedCentral
Metadata
Title
Alterations of p14 ARF , p15 INK4b , and p16 INK4a Genes in Primary Laryngeal Squamous Cell Carcinoma
Authors
Fernando López
Teresa Sampedro
José L. Llorente
Mario Hermsen
César Álvarez-Marcos
Publication date
01-01-2017
Publisher
Springer Netherlands
Published in
Pathology & Oncology Research / Issue 1/2017
Print ISSN: 1219-4956
Electronic ISSN: 1532-2807
DOI
https://doi.org/10.1007/s12253-016-0083-4

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