Immunohistochemical expression of P16 and cyclin D1 in oral lesions


  • José Ronildo Lins do Carmo Filho Universidade Federal do Ceará
  • Dennys Ramon de Melo Fernandes Universidade Federal do Ceará
  • Paulo Goberlânio Paulo Goberlânio de Barros Silva Universidade Federal do Ceará
  • Régia Maria do Socorro Vidal Patrocínio Universidade Federal do Ceará
  • Maria Elisa Quezado Lima Verde Universidade Federal do Ceará
  • Mário Rogério Mário Rogério Lima Mota Universidade Federal do Ceará
  • Fabrício Bitu Sousa Universidade Federal do Ceará
  • Ana Paula Negreiros Nunes Alves Universidade Federal do Ceará



Mouth Neoplasms, p16, Cyclin D1, Ki-67 Antigen


Aim: To evaluate the immunohistochemical expression of p16, cyclin D1 and Ki67 in oral lesions (fibroepithelial hyperplasia, squamous papilloma and epidermoid carcinoma). Study type: This is a retrospective, quantitative and cross-sectional study, carried out by the survey of paraffin blocks in the Oral Pathology Laboratory of the Dentistry course of the Federal University of Ceará, from 2008 to 2014. Methods: A total of 89 samples were selected, grouped into: 25 fibroepithelialhyperplasias (FHE); 16 oral squamous papillomas (OP); 28 oral squamous cell carcinomas (SCC), subdivided between patients below 50 years (SCC-50) and above 50 years (SCC+50). Twenty cases of oral mucosa without microscopic changes were used as the control group (CTL). Main outcome measures: Morphological evaluation through hematoxylin-eosin staining; Quantitative and dichotomous immunoexpression by streptavidin-biotin technique (P16, cyclin D1 and Ki67). Results: All evaluated groups, except for SCC+50 and CTL, presented a significantly higher frequency of cytological changes consistent with koilocytosis (p <0.001). The benign oral lesions (FHE and OP) and the SCC-50 group showed high immunostaining for p16 and cyclin D1, being statistically significant in relation to the control group (p <0.001). The quantitative expression of Ki67 was higher in SCC-50 than in the CTL group, with statistical significance (p <0.001). Conclusion: Oral lesions (OP, FHE and SCC-50) showed, together, koilocytosis and significant positivity for p16 and cyclin D1. Further studies should be conducted to clarify the possible relationship between cytopathic changes and p16 expression.


Doorbar J, Quint W, Banks L, Bravo IG, Stoler M, Broker TR, Stanley MA. The biology and life-cycle of human papillomaviruses. Vaccine 30. 2012 (Suppl 5): F55–F70.

Lewis JS. P16 Immunohistochemistry as a standalone test for risk stratification in oropharyngeal squamous cell carcinoma. Head Neck Pathol. 2012 (Suppl 1): S75–S82.

El-Mofty SK, Zhang MQ, Davila RM. Histologic identification of human papillomavirus (HPV)-related squamous cell carcinoma in cervical lymph nodes: a reliable predictor of the site of an occult head and neck primary carcinoma. Head Neck Pathol. 2008 (3): 163–168.

Geißler C, Tahtali A, Diensthuber M, Gassner D, Stöver T, Wagenblast J. The role of p16 expression as a predictive marker in HPV-positive oral SCCHN--a retrospective single-center study. Anticancer Res. 2013 Mar; 33(3): 913-6.

Bilyk OO, Pande NT, Buchynska LG. Analysis of p53, p16 (INK4a), pRb and Cyclin D1 expression and human papillomavirus in primary ovarian serous carcinomas. Exp Oncol. 2011 Sep; 33(3):150-6.

Gatta LB, Berenzi A, Balzarini P, Dessy E, Angiero F, Alessandri G, Gambino A, Grigolato P, Benetti A.Diagnostic implications of L1, p16, and Ki-67 proteins and HPV DNA in low-grade cervical intraepithelial neoplasia. Int J Gynecol Pathol. 2011 Nov; 30(6):597-604.

Wang HY, Kim G, Cho H, Kim S, Lee D, Park S, Park KH, Lee H. Diagnostic performance of HPV E6/E7, hTERT, and Ki67 mRNA RT-qPCR assays on formalin-fixed paraffin-embedded cervical tissue specimens from women with cervical cancer. Exp Mol Pathol. 2015 Jun; 98(3): 510-6.

Vasilescu F, Ceauşu M, Tănase C, Stănculescu R, Vlădescu T, Ceauşu Z. P63 and Ki-67 assessment in HPV-induced cervical neoplasia. Rom J Morphol Embryol. 2009; 50(3): 357-61.P53.

Xavier SD, Filho IB, Lancellotti CLP. Prevalência de achados sugestivos de papilomavírus humano (HPV) em biópsias de carcinoma espinocelular de cavidade oral e orofaringe: estudo preliminary. Rev. Bras. Otorrinolaringol. vol.71 no.4 São Paulo July/Aug. 2005

Kate C, Nicolas W. HPV mRNA and p16 detection as biomarkers for the improved diagnosis of cervical neoplasia. Cancer Epidemiol Biomarkers Prev. 2008 Oct; 17(10): 2536–2545.

Eleuterio JJ. Associação entre a carga viral de HPV de alto risco, expressão de p16INK4a e lesões intra-epiteliais escamosas do colo uterino. Rev. Assoc. Med. Bras. 2007 53(6).

Thavaraj S, Stokes A, Guerra E, Bible J, Halligan E, Long A, Okpokam A, Sloan P, Odell E, Robinson M. Evaluation of human papilomavírus testing for squamous cell carcinoma of the tonsil in clinical practice. J Clin Pathol. 2011 64(4): 308–312.

Duncan LD, Winkler M, Carlson ER, Heidel RE, Kang E, Webb D. P16 immunohistochemistry can be used to detect human papillomavirus in oral cavity squamous cell carcinoma. J Oral Maxillofac Surg. 2013 Aug; 71(8): 1367-75 pub 2013 May.

Chen ZW, Weinreb I, Kamel-Reid S, Perez-Ordoñez B. Equivocal p16 immunostaining in squamous cell carcinoma of the head and neck: staining patterns are suggestive of HPV status. Head Neck Pathol. 2012 Dec; 6(4): 422-9. pub 2012 Jul 17.

Mooren JJ, Gültekin SE, Straetmans JM, Haesevoets A, Peutz-Kootstra CJ, Huebbers CU, Dienes HP, Wieland U, Ramaekers FC, Kremer B, Speel EJ, Klussmann JP. Immunostaining is a strong indicator for high-risk-HPV-associated oropharyngeal carcinomas and dysplasias, but is unreliable to predict low-risk-HPV-infection in head and neck papillomas and laryngeal dysplasias. Int J Cancer. 2014 May 1; 134(9): 2108-17.

Doxtader EE, Katzenstein AL. The relationship between p16 expression and high-risk human papillomavirus infection in squamous cell carcinomas from sites other than uterine cervix: a study of 137 cases. Hum Pathol. 2012 Mar; 43(3): 327-32. doi: 10.1016/j.humpath.2011.05.010. Epub 2011 Aug 12.

Antonsson A, Neale RE, Boros S, Lampe G, Coman WB, Pryor DI, Porceddu SV, Whiteman DC. Human papillomavirus status and p16 (INK4A) expression in patients with mucosal squamous cell carcinoma of the head and neck in Queensland, Australia. Cancer Epidemiol. 2015 Apr; 39(2): 174-81. doi: 10.1016/j.canep.2015.01.010. Epub 2015 Feb 9.

Jour G, West K, Ghali V, Shank D, Ephrem BM. Differential Expression of p16INK4A and Cyclin D1 in Benign and Malignant Salivary Gland Tumors: A Study of 44 Cases. Head Neck Pathol. 2013 Sep; 7(3): 224–231 Published online 2013 Jan 13. doi: 10.1007/s12105-012-0417-9.

Angelo LA. P16INK4a expression correlates with degree of oral neoplasia: a comparison with Ki-67and Cyclin D1, expression and detection of high-risk HPV types. Mod Pathol. 2007; 16: 665–73

Ikenberg H, Bergeron C, Schmidt D, Griesser H, Alameda F, Angeloni C, Bogers J, Dachez R, Denton K, Hariri J, Keller T, von Knebel Doeberitz M, Neumann HH, Puig-Tintore LM, Sideri M, Rehm S, Ridder R; PALMS. Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study. J Natl Cancer Inst. 2013 Oct 16; 105(20): 1550-7.

Wentzensen N, Schwartz L, Zuna RE, Smith K, Mathews C, Gold MA, Allen RA, Zhang R, Dunn ST, Walker JL, Schiffman M.Performance of p16/Ki-67 immunostaining to detect cervical cancer precursors in a colposcopy referral population. Clin Cancer Res. 2012 Aug 1; 18(15): 4154-62. doi: 10.1158/1078-0432.CCR-12-0270. pub 2012 Jun 6.

Lassen P, Overgaard J. Scoring and classification of oropharyngeal carcinoma based on HPV-related p16-expression. Radiother Oncol. 2012 105(2): 269–270.




Como Citar

Lins do Carmo Filho JR, de Melo Fernandes DR, Paulo Goberlânio de Barros Silva PG, do Socorro Vidal Patrocínio RM, Quezado Lima Verde ME, Mário Rogério Lima Mota MR, Bitu Sousa F, Negreiros Nunes Alves AP. Immunohistochemical expression of P16 and cyclin D1 in oral lesions. RFPP [Internet]. 27º de setembro de 2023 [citado 17º de junho de 2024];3(3). Disponível em: