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Expression of p53 protein in laryngeal squamous cell carcinoma and dysplasia: possible correlation with human papillomavirus infection and clinicopathological findings

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Abstract

In order to evaluate the expression of p53 protein in 28 premalignant and 40 malignant squamous cell proliferations of the larynx and its relationship to tobacco consumption, human papillomavirus infection and differentiation grade of the lesions, p53 expression was examined by means of a microwave post-fixation immunohistochemical method using the PAb 240 and PAb 1801 monoclonal antibodies. HPV infection was assessed by non-isotopic in situ hybridization (NISH) and polymerase chain reaction (PCR). A large proportion of carcinomas (77.5%) and dysplasias (61%) expressed p53. No difference was found between differentiation grades of the lesions regarding p53 detection (P>0.1), but moderate or intense p53 expression was more frequent in the carcinomas (P<0.05). A statistical correlation was found between cigarette consumption and both p53 detection and p53 staining intensity (P<0.05 in each case). HPV study revealed HPV 16 and 18 infection only in carcinomas. The frequency was 28% and the physical state of the virus as demonstrated by NISH was integration into the genome. We observed an inverse relationship between HPV infection and p53 expression (P=0.006). Our findings suggest that p53 overexpression is a common and early event which increases in frequency with progression of laryngeal squamous cell carcinoma. The expression of p53 is influenced by tobacco and high-risk types of HPV.

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References

  1. Anwar K, Nakakuki K, Naiki H, Inuzuka M (1993) ras gene mutations and HPV infection are common in human laryngeal carcinoma. Int J Cancer 53:22–28

    Google Scholar 

  2. Anwar K, Nakakuki K, Imai H, Naiki H, Inuzuka M (1993) Over-expression of p53 protein in human laryngeal carcinoma. Int J Cancer 53:952–956

    Google Scholar 

  3. Banks L, Matlashewski G, Crawford L (1986) Isolation of human-p53-specific antibodies and their use in studies of human p53 expression. Eur J Biochem 159:529–534

    Google Scholar 

  4. Batsakis JG (1979) Tumors of the head and neck: clinical and pathological considerations, 2nd edn. Williams and Wilkins, Baltimore, p 574

    Google Scholar 

  5. Borresen AL, Helland A, Nesland J, Holm R, Trope C, Kaern J (1992) Papillomaviruses, p53 and cervical cancer. Lancet 339:1350–1351

    Google Scholar 

  6. Boshart M, Gissmann L, Ikenberg H, Kleinheinz A, Scheurlen W, ZurHausen H (1984) A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. EMBO J 3:1151–1157

    Google Scholar 

  7. Boyle JO, Hakim J, Koch W, Riet P van der, Hruban RH, Roa RA, Correo R, Eby YJ, Ruppert JM, Sidransky D (1993) The incidence of p53 mutations increases with progression of head and neck cancer. Cancer Res 53:4477–4480

    Google Scholar 

  8. Brandsma JL, Steinberg BM, Abramson AL, Winkler B (1986) Presence of human papilloma virus type 16 related sequences in verrucous carcinoma of the larynx. Cancer Res 46:2185–2188

    CAS  PubMed  Google Scholar 

  9. Braun L, Durst M, Mimuko R, Grupposo P (1990) Differential response of nontumorigenic and tumorigenic human papillomavirus type 16-positive epithelial cells to transforming growth factor β. Cancer Res 50:7324–7332

    Google Scholar 

  10. Cattoretti G, Pileri S, Parravicini C, Becker MHG, Poggi S, Bifulco C, Key G, D'amato L, Sabattini E, Feudale E, Reynolds F, Gerdes J, Rilke F (1993) Antigen unmasking on formalin-fixed, paraffin-embedded tissue sections. J Pathol 171:83–98

    Google Scholar 

  11. Cole ST, Streeck RE (1986) Genome organization and nucleotide sequence of human papillomavirus type 33, which is associated with cervical cancer. J Virol 58:991–995

    Google Scholar 

  12. Cooper K, Herrington CS, Graham AK, Evans MF, McGee JO'D (1991) In situ evidence for HPV 16, 18, 33 integration in cervical squamous cell cancer in Britain and South Africa. J Clin Pathol 44:406–409

    Google Scholar 

  13. Crook T, Wrede D, Tidy JA, Mason WP, Evans DJ, Vousden KH (1992) Clonal p53 mutation in primary cervical cancer: association with human-papillomavirus-negative tumours. Lancet 339:1070–1073

    Google Scholar 

  14. Dartmann K, Schwarz E, Gissmann L, ZurHausen H (1989) The nucleotide sequence and genome organization of human papillomavirus 11. Virology 151:124–130

    Google Scholar 

  15. Deb S, Jackson CT, Subler MA, Martin DW (1992) Modulation of cellular and viral promoters by mutant human p53 proteins found in tumor cells. J Virol 66:6164–6170

    Google Scholar 

  16. Dekmezian RH, Batsakis JG, Goepfert H (1987) In situ hybridization of papillomavirus DNA in head and neck squamous cell carcinomas. Arch Otolaryngol Head Neck Surg 113:819–821

    Google Scholar 

  17. Demers GW, Halbert CL, Galloway DA (1994) Elevated wild-type p53 protein levels in human epithelial cell lines immortalized by the human papillomavirus type 16 E6 gene. Virology 198:169–174

    Google Scholar 

  18. DeVilliers EM (1989) Heterogeneity of the human papillomavirus group. J Virol 63:4898–4903

    Google Scholar 

  19. Dolcetti R, Doglioni C, Maestro R, Gasparotto T, Barzan L, Pastore A, Romanelli M, Boiocchi M (1992) p53 overexpression is an early event in the development of human squamous cell carcinoma of the larynx; genetic and prognostic implications. Int J Cancer 52:178–182

    Google Scholar 

  20. Edmundson WF (1948) Microscopic grading of cancer and its practical implication. Arch Dermatol Syphilol 57:141–150

    Google Scholar 

  21. Field JK, Spandidos DA, Malliri A, Gosney JR, Yiagnisis M, Stell PM (1991) Elevated p53 expression correlates with a history of heavy smoking in squamous cell carcinoma of the head and neck. Br J Cancer 64:573–577

    Google Scholar 

  22. Fuzita M, Inoue M, Tanizawa O, Iwamoto S, Enomoto T (1992) Alterations of the p53 gene in human primary cervical carcinoma with and without human papillomavirus infection. Cancer Res 52:5223–5328

    Google Scholar 

  23. Gannon JV, Greaves R, Iggo R, Lane DP (1990) Activating mutations in p53 produce a common conformational effect. A monoclonal antibody specific for the mutant form. EMBO J 9:1595–1602

    Google Scholar 

  24. Gloss B, Bernard HV, Seedorf K, Klock G (1987) The up-stream regulatory region of human papilloma virus 16 contains an E2 protein-independent enhancer with is specific for cervical cells and regulated by glucocorticoid hormones. EMBO J 6:3735–3743

    Google Scholar 

  25. Gorgoulis V, Giatromanolaki A, Karameris A, Tsatsanis C, Aninos D, Ozanne B, Velsemes M, Jordanoglou J, Trigidou R, Papastamatiou H, Spandidos DA (1993) Epidermal growth factor receptor expression in squamous cell lung carcinomas: an immunohistochemical and gene analysis in formalin-fixed, paraffin-embedded material. Virchows Arch [A] 423:295–302

    Google Scholar 

  26. Hainaut P, Milner J (1992) Interaction of heat-shock protein 70 with p53 translated in vitro: evidence for interaction with dimeric p53 and for a role in the regulation of p53 conformation. EMBO J 11:3513–3520

    Google Scholar 

  27. Halevy O, Michalowitz D, Oren M (1990) Different tumor-derived p53 mutants exhibit distinct biological activities. Science 250:113–116

    Google Scholar 

  28. Harris AL (1990) Mutant p53 — the commonest genetic abnormally in human cancer; (editorial) J Pathol 162:5–6

    Google Scholar 

  29. Harris CC (1993) p53: At the crossroads of molecular carcinogenesis and risk assessment. Science 262:1980–1981

    Google Scholar 

  30. Hellquist H, Lundgren J, Olofsson J (1982) Hyperplasia, keratosis, dysplasia and carcinoma in situ of the vocal cords: a follow-up study. Clin Otolaryngol 7:11–27

    Google Scholar 

  31. Herrington CS, Burns J, Graham AK, Bhatt B, McGee JO'D (1989) Interphase cytogenetic using biotin and digoxigenin labelled probes. I. Relative sensitivity of both reporter molecules for the detection of HPV 16 in CaSki cells. J Clin Pathol 42:591–600

    Google Scholar 

  32. Hollstein MC, Metcalf RA, Welsh A, Montesano R, Harris CC (1990) Frequent mutation of the p53 gene in human esophageal cancer. Proc Natl Acad Sci USA 887:9958–9961

    Google Scholar 

  33. Hollstein MC, Peri L, Mandard AM, Wellsh JA, Montesano R, Metcalf RA, Bak M, Harris CC (1991) Genetic analysis of human esophageal tumors from two high incidence geographic areas: frequent p53 base substitutions and absence of ras mutations. CancerRes 51:4102–4106

    Google Scholar 

  34. Hong W, Yu-chun L, Xiu-shui K, Gang-zhong L, Jie L, Baoyin L, Li-juan Q, Ke-ring M (1991) Detection of human papillomarivus (HPV) in laryngeal carcinoma tissue. Chin Med J 104:523–525

    Google Scholar 

  35. Hoshikawa T, Nakajima T, Uhara H, Gotoh M, Shimosato Y, Tsutsumi K, Ono I, Ebihara S (1990) Detection of human papillomavirus DNA in laryngeal squamous cell carcinomas by polymerase chain reaction. Laryngoscope 100:450–647

    Google Scholar 

  36. Iggo R, Gatter K, Bartek J, Lane D, Harris AL (1990) Increased expression of mutant forms of p53 oncogene in primary lung cancer. Lancet 335:675–679

    Google Scholar 

  37. Kahn T, Schwarz E, ZurHausen H (1986) Molecular cloning and characterization of the DNA of a new human papillomavirus (HPV 30) from a laryngeal carcinoma. Int J Cancer 37:61–65

    Google Scholar 

  38. Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW (1991) Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51:6304–6311

    CAS  PubMed  Google Scholar 

  39. Levin AJ, Momand J, Finlay CA (1991) The p53 tumour suppressor gene. Nature 351:453–456

    Google Scholar 

  40. Lorincz AT, Lancaster WD, Temple GF (1986) Cloning and characterization of the DNA of a new human papillomavirus from a woman with dysplasia of the uterine cervix. J Virol 58:225–229

    Google Scholar 

  41. Lorincz AT, Quinn AP, Lancaster WD, Temple GP (1987) A new papillomavirus associated with cancer of the uterine cervix. Virology 159:187–190

    Google Scholar 

  42. Maestro R, Dolcetti R, Gasparotto D, Doglioni C, Pelucchi S, Barzan L, Grandi E, Boiocchi M (1992) High frequency of p53 gene alterations associated with protein overexpression in human squamous cell carcinoma of the larynx. Oncogene 7:1159–1166

    Google Scholar 

  43. McCoy DG, Wynder EL (1979) Etiological and preventive implications in alcohol carcinogenesis. Cancer Res 39:2844–2850

    Google Scholar 

  44. Mercer WE, Baserga R (1985) Expression of the p53 protein during the cell cycle of human peripheral blood lymphocytes. Exp Cell Res 160:31–46

    Google Scholar 

  45. Mercer WE, Shields MT, Lin D, Appela E, Ullrich SJ (1991) Growth suppression induced by wild-type p53 protein is accompanied by selective down-regulation of proliferating cell nuclear antigen expression. Proc Natl Acad Sci USA 88:1958–1962

    Google Scholar 

  46. Michaels L (1984) Pathology of the larynx, chapter 34. Squamous cell carcinoma: epidemilogy, early lesions, biopsy diagnosis. Springer, Berlin Heidelberg New York, pp 199–204

    Google Scholar 

  47. Milner J (1991) A conformational hypothesis for the suppressor and promoter functions of p53 in cell growth control and in cancer. Proc R Soc Lond [B] 245:139–145

    Google Scholar 

  48. Momand J, Zambetti GP, Olson DC, George D, Levine AJ (1992) The mdm-2 oncogene product forms a complex with p53 protein and inhibits p53-mediated transactivation. Cell 69:1237–1245

    Google Scholar 

  49. Morgan DW, Abdullah V, Quiney R, Myint S (1991) Human papilloma virus and carcinoma of the laryngopharynx. J Laryngol Otol 105:288–290

    Google Scholar 

  50. Mounts P, Shah K, Kashima H (1982) Viral etiology of juvenile- and adult-onset squamous papilloma of the larynx. Proc Natl Acad Sci USA 79:5425–5429

    Google Scholar 

  51. Perez-Ayala M, Ruiz-Cabello F, Esteban F, Concha A, Redondo M, Oliva MR, Cabrera T, Carrido F (1990) Presence of HPV 16 sequences in laryngeal carcinomas. Int J Cancer 46:8–11

    Google Scholar 

  52. Pfister H (1987) Human papillomaviruses and genital cancer. Adv Cancer Res 48:113–117

    Google Scholar 

  53. Rodrigues NR, Rowan A, Smith M et al. (1990) p53 mutations in colorectal cancer. Proc Natl Acad Sci USA 87:7555–7559

    Google Scholar 

  54. Scheffner M, Werness BA, Huibregste SM, Levine AJ, Howley PM (1990) The E6 oncoprotein encoded by HPV types 16 and 18 promotes the degradation of p53. Cell 63:1129–1136

    Google Scholar 

  55. Scheurlen W, Stremlau A, Gissmann L, Hohn D, Zenner H-P, ZurHausen H (1986) Rearranged HPV 16 molecules in an anal and in a laryngeal carcinoma. Int J Cancer 38:671–676

    Google Scholar 

  56. Schmidt W, Popham RE (1981) The role of drinking and smoking in mortality from cancer and other causes in male alcoholics. Cancer 47:1030–1041

    Google Scholar 

  57. Schneider A, Hotz M, Gissman L (1987) Increased prevalence of human papillomaviruses in the lower genital tract of pregnant women. Int J Cancer 40:198–201

    Google Scholar 

  58. Schuller DE, Abou-Issa H, Parrish R (1984) Estrogen and progesterone receptors in head and neck cancer. Arch Otolaryngol 110:725–727

    Google Scholar 

  59. Schwarz E, Durst M, Demankowski C et al. (1983) DNA sequence and genome organization of genital human papillomavirus type 6b. EMBO J 2:2341–2348

    Google Scholar 

  60. Seedorf K, Krammer G, Durst M, Suhai S, Rowekamp WG (1985) Human papillomavirus type 16 DNA sequence. Virology 145:181–185

    Google Scholar 

  61. Suler MA, Martin DW, Deb S (1992) Inhibition of viral and cellular promoters by human wild-type p53. J Virol 66:4757–4762

    Google Scholar 

  62. Syrjanen S, Syrajanen K, Mantyjarvi R, et al. (1987) Human papillomavirus DNA in squamous cell carcinomas of larynx demonstrated by in situ hybridization. ORL J Otorhinolaryngol Relat Spec 49:175–186

    Google Scholar 

  63. Tsutsumi K, Nakajima T, Gotoh M, et al. (1989) In situ hybridization and immunohistochemical study of human papillomavirus infection in adult laryngeal papillomas. Laryngoscope 99:80–85

    Google Scholar 

  64. Tuyns AJ (1991) Alcohol and cancer. An instructive association. Br J Cancer 64:415–416

    Google Scholar 

  65. Varley GM, Brammar W, Lane DP, Swallow JE, Dolan C, Walker RA (1991) Loss of chromosome 17p13 sequences and mutation of p53 in human breast carcinomas. Oncogene 6:413–421

    Google Scholar 

  66. Watts S, Brewer EE, Fry TL (1991) HPV DNA in head and neck carcinomas. Oral Surg Oral Med Oral Pathol 71:701–707

    Google Scholar 

  67. Wynder EL, Stellman SD (1977) Comparative epidemiology of tobacco-related cancers. Cancer Res 37:4608–4622

    Google Scholar 

  68. Wynford-Thomas D (1992) p53 in tumour pathology: can we trust immunochemistry? (editorial) J Pathol 166:329–330

    Google Scholar 

  69. Young LS, Bevan IS, Johnson MA, Blomfield PI, Maitland NJ, Woodman CBJ (1989). The polymerase chain reaction: a new epidemiological tool for investigating cervical human papillomavirus infection. BMJ 298:14–18

    Google Scholar 

  70. Zambetti GP, Levine AJ (1993) A comparison of the biological activities of wild-type and mutant p53. FASEB J 7:855–865

    Google Scholar 

  71. ZurHausen H (1989) Papillomaviruses in anogenital cancer as a model to understand the role of viruses in human cancers. Cancer Res 49:4677–4681

    Google Scholar 

  72. ZurHausen H (1991) Human papillomaviruses in the pathogenesis of anogenital cancer. Virology 184:9–13

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Histology and Embryology, University of Athens School of Medicine, Athens, Greece

    V. Gorgoulis, G. Rassidakis & C. Kittas

  2. Department of Pathology, “Red Cross” Hospital, Athens, Greece

    V. Gorgoulis, A. Giatromanolaki & C. Barbatis

  3. Department of Pathology, 401 General Army Hospital, Athens, Greece

    A. Karameris

  4. Antaiou 53, Lamprini, Ano Patisia, GR-11146, Athens, Greece

    V. Gorgoulis

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  1. V. Gorgoulis
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Gorgoulis, V., Rassidakis, G., Kittas, C. et al. Expression of p53 protein in laryngeal squamous cell carcinoma and dysplasia: possible correlation with human papillomavirus infection and clinicopathological findings. Vichows Archiv A Pathol Anat 425, 481–489 (1994). https://doi.org/10.1007/BF00197551

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