Journal of Cancer Research and Clinical Oncology

, Volume 129, Issue 11, pp 642–650 | Cite as

Differential alterations of the genes in the CDKN2A-CCND1-CDK4-RB1 pathway are associated with the development of head and neck squamous cell carcinoma in Indian patients

  • Anusri Tripathi (Bhar)
  • Soma Banerjee
  • Neelanjana Chunder
  • Anup Roy
  • Arunava Sengupta
  • Bidyut Roy
  • Susanta Roychowdhury
  • Chinmay Kumar Panda
Original Paper



The aim of this study was to analyse the alterations of the genes in the CDKN2A/CCND1/CDK4/RB1 pathway in the G1-S phase of the cell cycle during development of head and neck squamous cell carcinoma (HNSCC).


The alterations of these genes were analysed in 22 dysplastic lesions, 26 stage-I/II and 33 stage-III/IV HNSCC tumours of Indian patients.


The alterations [mutation, hypermethylation, homozygous deletion and loss of heterozygosity/microsatellite size alteration (LOH/MA)] in the CDKN2A were found to be highest in 57% of the samples, followed by CCND1 amplification and LOH/MA at the RB1 locus in 14% and 8.5% of the samples, respectively. No dominant CDK4 Arg24Cys mutation was seen in our samples. Comparatively high frequency of CDKN2A alterations (except homozygous deletion) was found in dysplastic head and neck lesions and remained almost constant or increased during progression of the tumour, whereas the homozygous deletion of CDKN2A and the alterations in CCND1 and RB1 genes were seen mainly in the later stages of the tumour.


Our study suggested that mutation/hypermethylation/allelic alterations (LOH/MA) of CDKN2A were associated with the development of dysplastic head and neck lesions. All the other alterations might provide some cumulative effect during progression of later stages of the tumour to have selective growth advantages.


Head and neck squamous cell carcinoma CDKN2A-CCND1-CDK4-RB1 Mutation Hypermethylation Homozygous deletion Loss of heterozygosity Microsatellite size alteration 



We are grateful to the Director of Chittaranjan National Cancer Institute (CNCI), Calcutta, Dr. S. Gupta, Director of the Cancer Centre and Welfare Home, Calcutta, and Drs. E. Zabarovsky, A Dam and S Mondal for their active support during this work. Financial support for this work was provided by grant no. BT/MB/05/002/94 from the Department of Biotechnology of the Government of India.


  1. Callender T, El-Naggar AK, Lee MS, Frankenthaler R, Luna MA, Batsakis JG (1994) Prad-1 (CCND1)/Cyclin D1 oncogene amplification in primary head and neck squamous cell carcinoma. Cancer 74:152–158PubMedGoogle Scholar
  2. Cerilli LA, Swartzbaugh JR, Saadut R, Marshall CE, Rumpel CA, Moskaluk CA Jr, Frierson H (1999) Analysis of chromosome 9p21 deletion and p16 mutation in salivary gland carcinomas. Hum Pathol 30:1242–1246PubMedGoogle Scholar
  3. Dasgupta S, Mukherjee N, Roy S, Roy A, Sengupta A, Roychowdhury S, Panda CK (2002) Mapping of the candidate tumor suppressor genes' loci on human chromosome 3 in head and neck squamous cell carcinoma of Indian patient population. Oral Oncol 38:6–15CrossRefPubMedGoogle Scholar
  4. Heyman M, Rasool O, Brandter LB, Liu Y, Grander D, Soderhall S, Gustavsson G, Einhorn S (1996) Prognostic importance of p15INK4B and p16INK4A gene inactivation in childhood acute lymphocytic leukemia. J Clin Oncol 14:1512–1520PubMedGoogle Scholar
  5. Kannan K, Munirajan AK, Krishnamurthy J, Bhuvarahamurthy V, Mohanprasad BK, Panishankar KH, Tsuchida N, Sanmugam G (2000) The p16 INK4 alpha/p19ARF gene mutations are infrequent and are mutually exclusive to p53 mutations in Indian oral squamous cell carcinomas. Int J Oncol 16:585–590PubMedGoogle Scholar
  6. Klangby BU, Okan I, Magnusson KP, Wendland M, Lind P, Wiman KG (1998) P16/INK4a and p15/INK4b gene methylation and absence of p16/INK4a mRNA and protein expression in Burkitt's lymphoma. Blood 91:1680–1687PubMedGoogle Scholar
  7. Knudson AG Jr (1995) Mutation and cancer: a personal odyssey. Adv Cancer Res 67:1–23PubMedGoogle Scholar
  8. Kyomoto R, Kumazawa H, Toda Y, Sakaida N, Okamura A, Inawaga M, Shintaku M, Yamashita T, Hiai H, Fukumoto M (1997) Cyclin-D1-gene amplification is a more potent prognostic factor than its protein overexpression in human head-and-neck squamous-cell carcinoma. Int J Cancer 74:576–581Google Scholar
  9. Li X, Lee NK, Ye YW, Waber PG, Schweitzer C, Cheng QC, Nisen PD (1994) Allelic loss at chromosome 3p, 8p, 13q and 17p associated with poor prognosis in head and neck cancer. J. Natl. Cancer Inst 86:1524–1529Google Scholar
  10. Liew CL, Li HM, Lo KW, Leow CK, Chan JY, Hin LY, Lau WY, Lai PBS, Lim BK, Huang J, Leung WT, Wu S, Lee JCK (1999) High frequency of p16INK4A gene alterations in hepatocellular carcinoma. Oncogene 18:789–795PubMedGoogle Scholar
  11. Lin SC, Chang KW, Chang CS, Liu TY, Tzeng YS, Yang FS, Wong YK (2000) Alterations of p16/MTS1 gene in oral squamous cell carcinomas from Taiwanese. J Oral Pathol Med 29:159–166CrossRefPubMedGoogle Scholar
  12. Lunderberg AS, Weinberg RA (1999) Control of the cell cycle and apoptosis. Eur J Cancer 35:531–539CrossRefPubMedGoogle Scholar
  13. MacGrogan D, Levy A, Bostwick D, Wagner M, Wells D, Bookstein R (1994) Loss of chromosome arm 8p loci in prostate cancer: mapping by quantitative allelic imbalance. Genes Chromosomes Cancer 10:151–159PubMedGoogle Scholar
  14. Mao L, El-Naggar AK (1999) Molecular changes in the multistage pathogenesis of head and neck cancer. In: Srivastava S, et al. (eds) Molecular pathology of early cancer. IOS Press, pp 189–205Google Scholar
  15. Matsuda H, Konishi N, Hiasa Y, Hayashi Y, Tsuzuki T, Tao M, Kitahori Y, Yoshioka N, Kirita T, Sugimura M (1996) Alterations of p16/CDKN2, p53 and ras genes in oral squamous cell carcinoma and premalignant lesions. J Oral Pathol Med 25:232–238PubMedGoogle Scholar
  16. Nagai MA (1999) Genetic alterations in head and neck squamous cell carcinomas. Braz J Med Biol Res 32:897–904PubMedGoogle Scholar
  17. Nakahara Y, Shintani S, Mihara M, Ueyama Y, Matsumura T (2001) High frequency of homozygous deletion and methylation of p16(INK4A) gene in oral squamous cell carcinomas. Cancer Lett 163:221–228CrossRefPubMedGoogle Scholar
  18. Papadimitrakopoulou V, Izzo J, Lippman SM, Lee JS, Fan YH, Clayman G, Ro JY, Hittelman WN, Lotan R, Hong WK, Mao L (1997) Frequent inactivation of p16INK4a in oral premalignant lesions. Oncogene 14:1799–1803CrossRefPubMedGoogle Scholar
  19. Park JS, Dong SM, Kim HS, Lee JY, Um SJ, Park IS, Kim SJ, Namkoong SE (1999) Detection of p16 gene alteration in cervical cancer using tissue microdissection and LOH study. Cancer Lett 136:101–108CrossRefPubMedGoogle Scholar
  20. Pollock PK, Pearson JV, Hayward NK (1996) Compilation of somatic mutations of the CDKN2 gene in human cancers: non-random distribution of base substitutions. Genes Chromosomes Cancer 15:77–88CrossRefPubMedGoogle Scholar
  21. Saranath D, Bhoite LT, Deo MG (1993) Molecular lesions in human oral cancer: the Indian scene. Oral Oncol 29B:107–112Google Scholar
  22. Schuyer M, Staveren I, Klijn JGM, Burg MEL, Stoter G, Henzen-Logmans SC, Foekens JA, Berns EMJJ (1996) Sporadic CDKN2A gene alterations in human ovarian tumors. Br J Cancer 74:1069–1073PubMedGoogle Scholar
  23. Shahnavaz SA, Bradley G, Regezi JA, Thakker N, Gao L, Hogg D, Jordan RC (2001) Patterns of CDKN2A gene loss in sequential oral epithelial dysplasias and carcinomas. Cancer Res 61:2371–2375PubMedGoogle Scholar
  24. 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 37:498–504CrossRefPubMedGoogle Scholar
  25. Tsai CH, Yang CC, Chou LS, Chou MY (2001) The correlation between alteration of p16 gene and clinical status in oral squamous cell carcinoma. J Oral Pathol Med 30:527–531CrossRefPubMedGoogle Scholar
  26. van der Riet P, Nawroz H, Hruban RH, Corio R, Tokino K, Koch W, Sidransky D (1994) Frequent loss of chromosome 9p21–22 early in head and neck cancer progression. Cancer Res 54:1156–1158PubMedGoogle Scholar
  27. Wu CL, Roz L, Mckown S, Sloan P, Read AP, Holland S, Porter S, Scully C, Paterson I, Tavasoli M, Thakker N (1999) DNA studies underestimate the major role of CDKN2A inactivation in oral and oropharyngeal squamous cell carcinomas. Genes Chromosomes Cancer 25:16–25CrossRefPubMedGoogle Scholar
  28. Yoo GH, Xu HJ, Brennan JA, Westra W, Hruban RH, Koch W, Benedict F, Sidransky D (1994) Infrequent inactivation of the retinoblastoma gene despite frequent loss of chromosome 13q in head and neck squamous cell carcinoma. Cancer Res 54:4603–4606PubMedGoogle Scholar
  29. Zabarovsky ER, Lerman MI, Minna JD (2002) Tumor suppressor genes on chromosome 3p involved in the pathogenesis of lung and other cancers. Oncogene 21:6915–6935CrossRefPubMedGoogle Scholar
  30. Zhang SY, Klein-Szanto AJP, Sauter ER, Safarenko M, Mitsunaga S, Nobori T, Carson DA, Ridge JA, Goodrow TL (1994) Higher frequency of alterations in the p16/CDKN2 gene in squamous cell carcinoma cell lines than in primary tumors of head and neck. Cancer Res 54:5050–5053Google Scholar
  31. Zuo L, Weger J, Yang Q, Goldstein AM, Tucker MA, Waker GJ, Hayward N, Dracopoli NC (1996) Germline mutations in the p16INK4a binding domain of CDK4 in familial melanoma. Nat Genet 12:97–99PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Anusri Tripathi (Bhar)
    • 1
  • Soma Banerjee
    • 2
  • Neelanjana Chunder
    • 1
  • Anup Roy
    • 3
  • Arunava Sengupta
    • 4
  • Bidyut Roy
    • 5
  • Susanta Roychowdhury
    • 2
  • Chinmay Kumar Panda
    • 1
  1. 1.Department of Oncogene RegulationChittaranjan National Cancer InstituteCalcutta 700026India
  2. 2.Department of Human Genetics and GenomicsIndian Institute of Chemical BiologyCalcutta 700032India
  3. 3.Department of PathologyBankura Sammilani Medical CollegeWest BengalIndia
  4. 4.Cancer Centre and Welfare HomeCalcutta 700063India
  5. 5.Anthropology and Human Genetics unitIndian Statistical InstituteCalcutta 700108India

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