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Aberrant promoter methylation and reduced expression of p16 gene in esophageal squamous cell carcinoma from Kashmir valley: a high-risk area

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Abstract

Esophageal squamous cell carcinoma (ESCC) is one of the most prevalent cancer in Jammu and Kashmir region of India and has multi-factorial etiology involving dietary habits, genetic factors, and gene environmental interactions. Inactivation of the p16 gene expression by aberrant promoter methylation plays an important role in the progression of esophageal carcinoma. In the present investigation, we have studied the role of p16 promoter methylation in 69 histopathologically confirmed ESCC tissues and compared it with corresponding normal adjacent tissues for DNA methylation in the CpG island in the p16 promoter region by methylation-specific polymerase chain reaction (MSP) and p16 protein expression by immunoblotting. The results showed loss of p16 expression in 67% (46/69) of tumor tissues compared to only 3% in control tissues (2/69). Promoter methylation was observed in 52% (36/69) of tumor tissues and it gradually increased with the increasing severity of histological grades of the cancer (P = 0.0001). Loss of p16 expression with promoter methylation was observed in 26 of 36 cases (72%). Analysis of patients dietary habits revealed a strong association between promoter methylation and high consumption of hot salted tea (P < 0.05) which is a most favourite drink commonly consumed by Kashmiri people.

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References

  1. Parkin DM, Bray F, Ferlay J, Pisani P (2005) Global cancer statistics, 2002. CA Cancer J Clin 55:74–108. doi:10.3322/canjclin.55.2.74

    Article  PubMed  Google Scholar 

  2. Phukan RK, Ali MS, Chetia CK, Mahanta J (2001) Betel nut and tobacco chewing; potential risk factors of cancer of oesophagus in Assam, India. Br J Cancer 85:661–667. doi:10.1054/bjoc.2001.1920

    Article  CAS  PubMed  Google Scholar 

  3. Khuroo MS, Zargar SA, Mahajan R, Banday MA (1992) High incidence of oesophageal and gastric cancer in Kashmir in a population with special personal and dietary habits. Gut 33:11–15. doi:10.1136/gut.33.1.11

    Article  CAS  PubMed  Google Scholar 

  4. Kumar R, Mende P, Wacker CD, Spiegelhalder B, Preussmann R, Siddiqi M (1992) Caffeine-derived N-nitroso compounds–I: nitrosatable precursors from caffeine and their potential relevance in the etiology of oesophageal and gastric cancers in Kashmir, India. Carcinogenesis 13:2179–2182. doi:10.1093/carcin/13.11.2179

    Article  CAS  PubMed  Google Scholar 

  5. Mir MM, Dar NA, Gochhait S, Zargar SA, Ahangar AG, Bamezai RN (2005) p53 Mutation profile of squamous cell carcinomas of the esophagus in Kashmir (India): a high-incidence area. Int J Cancer 116:62–68. doi:10.1002/ijc.21002

    Article  CAS  PubMed  Google Scholar 

  6. Katiyar S, Hedau S, Jain N et al (2005) p53 Gene mutation and human papillomavirus (HPV) infection in esophageal carcinoma from three different endemic geographic regions of India. Cancer Lett 218:69–79. doi:10.1016/j.canlet.2004.09.003

    Article  CAS  PubMed  Google Scholar 

  7. Murtaza I, Mushtaq D, Margoob MA, Dutt A, Wani NA, Ahmad I, Bhat ML (2006) A study on p53 gene alterations in esophageal squamous cell carcinoma and their correlation to common dietary risk factors among population of the Kashmir valley. World J Gastroenterol 12:4033–4037

    CAS  PubMed  Google Scholar 

  8. Jen J, Harper JW, Bigner SH, Bigner DD, Papadopoulos N, Markowitz S, Willson JK, Kinzler KW, Vogelstein B (1994) Deletion of p16 and p15 genes in brain tumors. Cancer Res 54:6353–6358

    CAS  PubMed  Google Scholar 

  9. Kamb A, Gruis NA, Weaver-Feldhaus J et al (1994) A cell cycle regulator potentially involved in genesis of many tumor types. Science 264:436–440. doi:10.1126/science.8153634

    Article  CAS  PubMed  Google Scholar 

  10. Cairns P, Polascik TJ, Eby Y et al (1995) Frequency of homozygous deletion at p16/CDKN2 in primary human tumours. Nat Genet 11:210–212. doi:10.1038/ng1095-210

    Article  CAS  PubMed  Google Scholar 

  11. Myohanen SK, Baylin SB, Herman JG (1998) Hypermethylation can selectively silence individual p16ink4A alleles in neoplasia. Cancer Res 58:591–593

    CAS  PubMed  Google Scholar 

  12. Lam AKY (2000) Molecular biology of esophageal squamous cell carcinoma. Crit Rev Oncol Hematol 33:71–90. doi:10.1016/S1040-8428(99)00054-2

    Article  CAS  PubMed  Google Scholar 

  13. Smeds J, Berggren P, Ma X, Xu Z, Hemminki K, Kumar R (2002) Genetic status of cell cycle regulators in squamous cell carcinoma of the oesophagus: the CDKN2A (p16(INK4a) and p14(ARF)) and p53 genes are major targets for inactivation. Carcinogenesis 23:645–655. doi:10.1093/carcin/23.4.645

    Article  CAS  PubMed  Google Scholar 

  14. Ralhan R, Mathew R, Arora S, Bahl R, Shukla NK, Mathur M (2000) Frequent alterations in the expression of tumor suppressor genes p16INK4A and pRb in esophageal squamous cell carcinoma in the Indian population. J Cancer Res Clin Oncol 126:655–660. doi:10.1007/s004320000147

    Article  CAS  PubMed  Google Scholar 

  15. Mori T, Miura K, Aoki T, Nishihira T, Mori S, Nakamura Y (1994) Frequent somatic mutation of the MTS1/CDK4I (multiple tumor suppressor/cyclin-dependent kinase 4 inhibitor) gene in esophageal squamous cell carcinoma. Cancer Res 54:3396–3397

    CAS  PubMed  Google Scholar 

  16. Xing EP, Nie Y, Song Y, Yang GY, Cai YC, Wang LD, Yang CS (1999) Mechanisms of inactivation of p14ARF, p15INK4b, and p16INK4a genes in human esophageal squamous cell carcinoma. Clin Cancer Res 5:2704–2713

    CAS  PubMed  Google Scholar 

  17. Abbaszadegan MR, Raziee HR, Ghafarzadegan K, Shakeri MT, Afsharnezhad S, Ghavamnasiry MR (2005) Aberrant p16 methylation, a possible epigenetic risk factor in familial esophageal squamous cell carcinoma. Int J Gastrointest Cancer 36:47–54. doi:10.1385/IJGC:36:1:047

    Article  CAS  PubMed  Google Scholar 

  18. Hayashi K, Metzger R, Salonga D et al (1997) High frequency of simultaneous loss of p16 and p16beta gene expression in squamous cell carcinoma of the esophagus but not in adenocarcinoma of the esophagus or stomach. Oncogene 15:1481–1488. doi:10.1038/sj.onc.1201295

    Article  CAS  PubMed  Google Scholar 

  19. Xing EP, Nie Y, Wang LD, Yang GY, Yang CS (1999) Aberrant methylation of p16INK4a and deletion of p15INK4b are frequent events in human esophageal cancer in Linxian, China. Carcinogenesis 20:77–84. doi:10.1093/carcin/20.1.77

    Article  CAS  PubMed  Google Scholar 

  20. Tokugawa T, Sugihara H, Tani T, Hattori T (2002) Modes of silencing of p16 in development of esophageal squamous cell carcinoma. Cancer Res 62:4938–4944

    CAS  PubMed  Google Scholar 

  21. Ito S, Ohga T, Saeki H, Watanabe M, Kakeji Y, Morita M, Yamada T, Maehara Y (2007) Promoter hypermethylation and quantitative expression analysis of CDKN2A (p14ARF and pl6INK4a) gene in esophageal squamous cell carcinoma. Anticancer Res 27:3345–3353

    CAS  PubMed  Google Scholar 

  22. Fujiwara S, Noguchi T, Takeno S, Kimura Y, Fumoto S, Kawahara K (2008) Hypermethylation of p16 gene promoter correlates with loss of p16 expression that results in poorer prognosis in esophageal squamous cell carcinomas. Dis Esophagus 21:125–131. doi:10.1111/j.1442-2050.2007.00735.x

    Article  CAS  PubMed  Google Scholar 

  23. Coia LR, Myerson RJ, Tepper JE (1995) Late effects of radiation therapy on the gastrointestinal tract. Int J Radiat Oncol Biol Phys 31:1213–1236. doi:10.1016/0360-3016(94)00419-L

    CAS  PubMed  Google Scholar 

  24. Esophagus (1992) In: American Joint Committee on Cancer (ed) Manual for the staging of cancer. JB Lippincott, Philadelphia, pp 57–59

  25. Das BC, Sharma JK, Gopalakrishna V, Luthra UK (1992) Analysis by polymerase chain reaction of the physical state of human papillomavirus type 16 DNA in cervical preneoplastic and neoplastic lesions. J Gen Virol 73(Pt 9):2327–2336. doi:10.1099/0022-1317-73-9-2327

    Article  CAS  PubMed  Google Scholar 

  26. 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 USA 93:9821–9826. doi:10.1073/pnas.93.18.9821

    Article  CAS  PubMed  Google Scholar 

  27. Mishra A, Bharti AC, Varghese P, Saluja D, Das BC (2006) Differential expression and activation of NF-kappaB family proteins during oral carcinogenesis: role of high risk human papillomavirus infection. Int J Cancer 119:2840–2850. doi:10.1002/ijc.22262

    Article  CAS  PubMed  Google Scholar 

  28. Eads CA, Lord RV, Kurumboor SK et al (2000) Fields of aberrant CpG island hypermethylation in Barrett’s esophagus and associated adenocarcinoma. Cancer Res 60:5021–5026

    CAS  PubMed  Google Scholar 

  29. Reed JA, Loganzo F Jr, Shea CR et al (1995) Loss of expression of the p16/cyclin-dependent kinase inhibitor 2 tumor suppressor gene in melanocytic lesions correlates with invasive stage of tumor progression. Cancer Res 55:2713–2718

    CAS  PubMed  Google Scholar 

  30. Geradts J, Wilson PA (1996) High frequency of aberrant p16(INK4A) expression in human breast cancer. Am J Pathol 149:15–20

    CAS  PubMed  Google Scholar 

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Acknowledgments

The work was supported by institutional grants from Indian Council of Medical Research (ICMR), New Delhi. Fellowship support to IS by Sher-I-Kashmir Institute of Medical Science, Kashmir, India and to SH by ICMR. This work is carried out by IS and SH—who collected samples, and performed all experiments and primary manuscript writing.

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Author notes

  1. Bhudev C. Das

    Present address: Dr. B.R. Ambedkar Research Centre for Biomedical Research (ACBR), University of Delhi (North Campus), Delhi, 110007, India

Authors and Affiliations

  1. Department of Clinical Biochemistry, Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India

    Irfana Salam, Mohammad Muzaffar Mir, Nazir Ahmad Dar & Safiya Abdullah

  2. Division of Molecular Oncology, Institute of Cytology and Preventive Oncology (ICMR), I-7, Sector-39, Noida, India

    Showket Hussain, Suresh Hedau, Alok Chandra Bharti & Bhudev C. Das

  3. Department of Immunology and Molecular Medicine, Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India

    Mushtaq Ahmad Siddiqi

  4. Department of Cardiovascular Thoracic Surgery, Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India

    Riyaz Ahmad Lone

  5. Department of Gastroenterology, Sher-I-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India

    Showkat Ahmad Zargar

  6. Department of Statistics, Institute of Cytology and Preventive Oncology (ICMR), I-7, Sector-39, Noida, 201301, India

    Shashi Sharma

  7. Department of Biosciences, Jamia Millia Islamia, New Delhi, India

    Seemi Farhat Basir

Authors
  1. Irfana Salam
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  2. Showket Hussain
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  3. Mohammad Muzaffar Mir
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  8. Showkat Ahmad Zargar
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  9. Shashi Sharma
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  10. Suresh Hedau
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  11. Seemi Farhat Basir
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  13. Bhudev C. Das
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Corresponding author

Correspondence to Bhudev C. Das.

Additional information

Irfana Salam and Showket Hussain contributed equally to this paper.

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Salam, I., Hussain, S., Mir, M.M. et al. Aberrant promoter methylation and reduced expression of p16 gene in esophageal squamous cell carcinoma from Kashmir valley: a high-risk area. Mol Cell Biochem 332, 51–58 (2009). https://doi.org/10.1007/s11010-009-0173-7

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  • DOI: https://doi.org/10.1007/s11010-009-0173-7

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