Abstract
To examine the prognostic significance of the immunohistochemical expression of p63 and Ki-67 oncoproteins in patients with laryngeal squamous cell carcinoma, a retrospective evaluation was carried out on a cohort of 108 patients with primary laryngeal squamous cell carcinoma (LSCC) treated by primary surgery. For the immunohistochemical evaluation, tissue section obtained by formalin-fixed and paraffin-embedded tissue blocks from resection of each patient was used. Clinicopathologic data were associated with the immunostaining results. The association among the considered variables was assessed by Fisher’s exact test, Mann–Whitney test, non-parametric χ 2 test, and Spearman’s rho rank test was used to assess the relations among them. Differences in p63 and Ki-67 immunoreactivity among the different groups were compared via Kruskal–Wallis test and post hoc tests were performed using Mann–Whitney test with Bonferroni correction. The overall survival rate was estimated via Kaplan–Meier method, and the cumulative incidence functions for different groups were compared using log-rank statistics. Cox proportional hazard model was employed in a multivariate analysis to assess the effect of prognostic factors in the overall survival rate. Furthermore, taking into account death due to other causes, we estimated LSCC-related survival and disease-free survival rates using competing risk analysis. The results of immunohistochemical examination showed a statistically significant relationship between the up-regulation of P63 and Ki-67, an increase in histological grading, and primary tumours associated with lymph node metastases. p63 and Ki-67 up-regulation was related to a shorter disease-free survival and a significant association was found between p63 and Ki-67 percentage of positive cells and patient survival. Finally, we noticed a significant relation between p63 and Ki-67 (ρ = 0.87). On the other hand, no statistically significant associations were found between p63 and Ki-67 down-regulation and clinicopathologic data. Our findings suggest that abnormal p63 and Ki-67 immunoreactivity may be involved in the early phases of laryngeal tumorigenesis and may become a significant prognostic predictor for both overall and disease-free survivals. These biomarkers could thus help in the selection of high-risk patients with LSCC who may benefit from more aggressive therapy or chemoprevention.
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References
Taxy JB (1996) Upper respiratory tract. In: Damjanov I, Linder J (eds) Anderson’s pathology, 10th edn. Mosby, St. Louis, p 1463
Pradier R, Gonzalez A, Matos E, Loria D, Adan R, Saco P, Califano L (1993) Prognostic factors in laryngeal carcinoma: experience in 296 male patients. Cancer 71:2472–2476
Pera E, Moreno A, Galindo L (1986) Prognostic factors in laryngeal carcinoma: a multifactorial study of 416 cases. Cancer 58:928–934
Vlachtis K, Nikolau A, Markou K, Fountzilas G, Daniilidis I (2005) Clinical and molecular prognostic factors in operable laryngeal cancer. Eur Arch Otorhinolaryngol 262(11):890–898
Gallo O, Libonati GA, Gallina E, Fini-Storchi O, Giannini A, Urso C, Bondi R (1991) Langerhans cells related to prognosis in patients with laryngeal carcinoma. Arch Otolaryngol Head Neck Surg 117:1007–1010
Gerdes J, Schwab U, Lemke H et al (1983) Production of a monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer 31:13–20
Brown DC, Gatter KC (1990) Monoclonal antibody KI-67: its use in histopathology. Histopathology 17:489–503
Wilson GD, Saunders MI, Dische S et al (1996) Direct comparison of bromodeoxyuridine and Ki-67 labelling indices in human tumours. Cell Prolif 29:141–152
Levrero M, De Laurenzi V, Costanzo A, Gong J, Wang JY, Melino G (2000) The p53/p63/p73 family of transcription factors : overlapping and distinct functions. J Cell Sci 113:1661–1670
Nishi H, Isaka K, Sagawa Y, Ususa S, Fujito A, Ito H, Senoo M, Kato H, Takayama M (1999) Mutation and transcription analyses of the p63 gene in cervical carcinoma. Int J Oncol 15:1149–1153
Tani M, Shimizu K, Kawahara C, Kohno T, Ishimoto O, Ikawa S, Yokota J (1999) Mutation and expression of the p51 gene in human lung cancer. Neoplasia 1:71–79
Hagiwara K, McMenamin MG, Miura K, Harris CC (1999) Mutation analysis of the p63/p73L/p51/p40/CUSP/KET gene in human cancer cell lines using intronic primers. Cancer Res 59:4165–4169
Barbareschi M, Pecciarini L, Cangi MG, Macri E, Rizzo A, Viale G, Doglioni C (2001) P63, a p53 homologue, is a selective nuclear marker of myoepithelial cells of the human breast. Am J Surg Pathol 25:1054–1060
Glickman JN, Yang A, Shahsafaei A, McKeon F, Odze RD (2001) Expression of p53-related protein p63 in the gastrointestinal tract and in esophageal metaplastic and neoplastic disorders. Hum Pathol 32:1157–1165
Park BJ, Lee SJ, Kim JI, Lee SJ, Lee CH, Chang SG, Park JH, Chi SG (2000) Frequent alteration of p63 expression in human primary bladder carcinomas. Cancer Res 60:3370–3774
Yamaguchi K, Wu L, Caballero OL, Hibi K, Trink B, Resto V, Cairns P, Okami K, Koch WM, Sidransky D, Jen J (2000) Frequent gain of the p40/p51/p63 gene locus in primary head and neck squamous cell carcinoma. Int J Cancer 86:684–689
Lo Muzio L, Santarelli A, Caltabiano R, Rubini C, Pieramici T, Trevisiol L, Carinci F, Leonardi R, De Lillo A, Lanzafame S, Bufo P, Piattelli A (2005) P63 overexpression associates with poor prognosis in head and neck squamous cell carcinoma. Hum Pathol 36(2):187–194
Jia H, Wang J, Shan Y, Hou J (2006) The clinical significance of the expression of p63 gene in laryngeal squamous cell carcinoma. Lin Chuang Er Bi Yan Hou Ke Za Zhi 20(3):119–121
de Oliveira LR, Ribeiro-Silva A, Zucoloto S (2007) Prognostic impact of p53 and p63 immunoexpression in oral squamous cell carcinoma. J Oral Pathol Med 36(4):191–197
Cardesa A, Gale N, Nadal A, Zidar N (2005) Squamous cell carcinoma. In: Head and Neck tumors: Pathology & Genetics. World Health Organization Classification on Tumors (WHO), WHO press, Geneva, pp 118–119
Edge SB, Compton CC (2010) The American Joint Committee on Cancer: the 7th edition of the AJCC Cancer Staging Manual and the future of TNM. Ann Surg Oncol 2010(17):1471–1474
Fine JP, Gray RJ (1999) A proportional hazards model for the subdistribution of a competing risk. JASA 94:496–509
Gray RJ (1988) A class of K-sample tests for comparing the cumulative incidence of a competing risk. Ann Stat 16:1141–1154
R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, http://www.R-project.org/ (R program)
Langer CJ (2012) Exploring biomarkers in head and neck cancer. Cancer 118(16):3882–3892
Shin DM, Khuri FR (2013) Advances in the management of recurrent or metastatic squamous cell carcinoma of the head and neck. Head Neck 35(3):443–453
Re M, Magliulo G, Ferrante L, Zizzi A, Santarelli A, Stramazzotti D, Lo Muzio L, Goteri G, Rubini C (2013) p63 expression in laryngeal squamous cell carcinoma is related to tumor extension, histologic grade, lymph node involvement and clinical stage. J Biol Regul Homeost Agents 27(1):121–129
Re M, Magliulo G, Salvolini E, Orciani M, Gioacchini FM, Goteri G, Rubini C (2010) Prognostic significance of p53 and KAI-1 expression in patients with Laryngeal squamous cell carcinoma. Anal Quant Cytol Histol 32(5):247–253
Artico M, Bianchi E, Magliulo G, De Vincentis M, De Santis E, Orlandi A, Santoro A, Pastore FS, Giangaspero F, Caruso R, Re M, Fumagalli L (2012) Neurotrophins, their receptors and KI-67 in human GH secreting pituitary adenomas: an immunohistochemical analysis. Int J Imunopathol Pharmacol 25(1):117–125
Xie X, Clausen OP, De Angelis P, Boysen M (1999) The prognostic value of spontaneous apoptosis, Bax, Bcl-2, and p53 in oral squamous cell carcinoma of the tongue. Cancer 86:913–920
Re M, Magliulo G, Tarchini P, Mallardi V, Rubini C, Santarelli A, Lo Muzio L (2011) p53 and BCL-2 over-expression on inversely correlate with histological differentiation in occupational ethmoidal intestinal type sinonasal adenocarcinoma. Int J Imunopathol Pharmacol 24:603–609
Nylander K, Dabelsteen E, Hall PA (2000) The p53 molecule and its role in squamous cell carcinomas of the head and neck. J Oral Pathol Med 29:413–425
Lo Muzio L, Santarelli A, Caltabiano R et al (2005) p63 overexpression associates with poor prognosis in head and neck squamous cell carcinoma. Hum Pathol 36:187–194
Lo Muzio L, Campisi G, Farina A et al (2007) Effect of p63 expression on survival in oral squamous cell carcinoma. Cancer Investig 25:466–469
Ramer N, HaiShan W, Sabo E, Ramer Y, Emanuel P, Orta L, Burstein DE (2010) Prognostic value of quantitative p63 immunostaining in adenoid cystic carcinoma of salivary gland assessed by computerized image analysis. Cancer 1:77–83
Asioli S, Righi A, Volante M, Eusebi V, Bussolati G (2007) p63 Expression a new prognostic marker in Merkel cell carcinoma. Cancer 110:640–647
Rocco JW, Leong CO, Kuperwasser N et al (2006) p63 mediates survival in squamous cell carcinoma by suppression of p73-dependent apoptosis. Cancer Cell 9:45–56
Zangen R, Ratovitski E, Sidransky D (2005) DeltaNp63alpha levels correlate with clinical tumour response to cisplatin. Cell Cycle 4:1313–1315
Takahashi Y, Noguchi T, Takeno S, Kimura Y, Okubo M, Kawahara K (2006) Reduced expression of p63 has prognostic implications for patients with esophageal squamous cell carcinoma. Oncol Rep 15:323–328
Urist MJ, DiComo CJ, Lu ML et al (2002) Loss of p63 expression is associated with tumor progression in bladder cancer. Am J Pathol 161:1199–2006
Koga F, Kawakami S, Fujii Y et al (2003) Impaired p63 expression associates with poor prognosis and uroplakin III expression in invasive urothelial carcinoma of the bladder. Clin Cancer Res 9:5501–5507
Pruneri G, Pignataro L, Manzotti M, Carboni N, Ronchetti D, Neri A, Cesana M, Viale G (2002) p63 in laryngeal squamous cell carcinoma: evidence for a role of TA-p63 down regulation in tumorigenesis and lack of prognostic implications of p63 immunoreactivity. Lab Invest 82(10):1327–1334
Dong P, Li X, Zhu Z, Yu Z, Lu G, Sun Z, Wang S (2005) Expression of S-100 positive dendritic cell, TIMP-1 and p63 and its significance in the primary laryngeal carcinoma. Lin Chuang Er Bi Yan Hou Ke Za Zhi 19(3):311–322
Lazaris AC, Rigopoulou A, Tseleni-Balafouta S, Kavantzas N, Thimara I, Zorzos HS, Eutychiadis CA, Petraki K, Kandiloros D, Davaris P (2002) Immunodetection and clinico-pathological correlates of two tumour growth regulators in laryngeal carcinoma. Histol Histopathol 17:131–138
Wintzer HO, Zipfel I, Schulte-Mönting J, Hellerich U, von Kleist S (1991) Ki-67 immunostaining of human breast tumors and its relationship to prognosis. Cancer 67:421–428
Tungekar MF, Gatter KC, Dunnill MS, Masan DY (1991) Ki-67 immunostaining and survival in operable lung cancer. Histopathology 19:545–550
Youssef EM, Matsuda T, Takada N, Osugi H, Higashino M, Kinoshita H, Watanabe T, Katsura Y, Wanibushi H, Fukushima S (1995) Prognostic significance of the MIB-1 proliferation index for patients with squamous cell carcinoma of the esophagus. Cancer 76:358–366
Aaltomaa S, Lipponen P, Ala-Opas M, Eskelinen M, Syrjanen K (1997) Prognostic value of Ki-67 expression in renal cell carcinomas. Eur Urol 31:350–355
Holte H, de Lange DC, Beiske K, Stokke T, Marton PF, Smeland EB, Hoie J, Kvaloy S (1989) Ki-67 and 4F2 antigen expression as well as DNA synthesis predict survival at relapse/tumor progression in low-grade B-cell lymphoma. Int J Cancer 44:975–980
Golusinski W, Olofsson J, Szmeja Z, Majewski P (1996) P53, PCNA and Ki-67 in laryngeal cancer. Pol J Pathol 47:175–182
Perez Carro Rios A, Garcia Caballero T, Lozano Ramirez A, Arias Santos MD, Soto Varela A, Forteza Vila J, Labella Caballero T (2000) The relationship between survival and cell proliferation index in squamous cell carcinoma of larynx. Rev Laryngol Otol Rhinol (Bord) 121:121–125
Sittel C, Eckel HE, Damm M, von Pritzbuer E, Kvasnicka HM (2000) Ki-67 (MIB-1), p53, and Lewis-X (LeuM1) as prognostic factors of recurrence in T1 and T2 laryngeal carcinoma. Laryngoscope 110:1012–1017
Grzanka A, Sujkowska R, Janiak A, Adamska M (2000) Immunogold labelling of PCNA and Ki-67 antigen at the ultra-structural level in laryngeal squamous cell carcinoma and its correlation with lymph node metastasis and histological grade. Acta Histochem 102:139–149
Hagedorn HG, Tubel J, Wiest I, Nerlich AG (1998) In situ apoptotic and proliferation index in laryngeal squamous cell carcinomas. Anal Cell Pathol 16:177–184
Pulkkinen JO, Penttinen M, Jalkanen M, Klemi P, Grenman R (1997) Syndecan-1: a new prognostic marker in laryngeal cancer. Acta Otolaryngol 117:312–315
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Re, M., Zizzi, A., Ferrante, L. et al. p63 and Ki-67 immunostainings in laryngeal squamous cell carcinoma are related to survival. Eur Arch Otorhinolaryngol 271, 1641–1651 (2014). https://doi.org/10.1007/s00405-013-2833-1
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DOI: https://doi.org/10.1007/s00405-013-2833-1