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Human papillomavirus in oral cavity and oropharynx carcinomas in the central region of Brazil Please cite this article as: Petito G, Carneiro MA, Santos SH, Silva AM, Alencar RC, Gontijo AP. Human papillomavirus in oral cavity and oropharynx carcinomas in the central region of Brazil. Braz J Otorhinolaryngol. 2017;83:38-44.

Abstract

Introduction

Molecular studies about carcinomas of the oral cavity and oropharynx demonstrate the presence of human papilomavirus genome in these tumors, reinforcing the participation of human papilomavirus in oral carcinogenesis.

Objectives

This study aimed to determine the prevalence of human papilomavirus and genotype distribution of HPV16 and HPV18 in oral cavity and oropharynx carcinomas, as well as their association with clinical characteristics of the tumors.

Methods

This is a retrospective study, with clinical data collected from 82 patients. Human papilomavirus detection was conducted on specimens of oral cavity and oropharynx carcinomas included in paraffin blocks. Patients were assisted in a cancer reference center, in the central region of Brazil, between 2005 and 2007. Polymerase chain reaction was used for the detection and genotyping of human papilomavirus.

Results

Among the patients evaluated, 78% were male. The average age of the group was about 58 years. Risk factors, such as smoking (78%) and alcohol consumption (70.8%) were recorded for the group. HPV DNA was detected in 21 cases (25.6%; 95% confidence interval 16.9–36.6) of which 33.3% were HPV16 and 14.3% were HPV18. The presence of lymph node metastases and registered deaths were less frequent in human papilomavirus positive tumors, suggesting a better prognosis for these cases; however, the differences between the groups were not statistically significant.

Conclusion

The results obtained in the present study, with respect to the presence of the high-risk HPV16 and HPV18 genotypes, highlight the importance of human papilomavirus vaccination in the control of oral cavity and oropharynx carcinomas.

Keywords
Papillomaviridae; Papillomaviridae 16; Head and neck neoplasm; Epidemiology

Resumo

Introdução

Estudos moleculares sobre carcinomas da cavidade oral e orofaringe demonstram a presença do genoma do papilomavírus humano (HPV) nesses tumores, o que enfatiza a participação do HPV na carcinogênese oral.

Objetivos

Determinar a prevalência de HPV e a distribuição genotípica de HPV16 e HPV18 nos carcinomas de cavidade oral e orofaringe, bem como sua associação com as características clínicas dos tumores.

Método

Estudo retrospectivo, com dados clínicos coletados de 82 pacientes. A detecção de HPV foi feita em amostras de carcinomas de cavidade oral e orofaringe incluídos em blocos de parafina. Os pacientes foram atendidos em um centro de referência para tratamento do câncer, na região central do Brasil, entre 2005 e 2007. Foi usada a reação em cadeia de polimerase (PCR) para a detecção e genotipagem do HPV.

Resultados

Entre os pacientes avaliados, 78% eram homens. A média de idade do grupo era de 58 anos. Fatores de risco como o tabagismo (78%) e consumo de álcool (70,8%) foram registrados para o grupo. HPV DNA foi detectado em 21 casos (25,6%; IC de 95%, 16,9-36,6), dos quais 33,3% eram HPV16 e 14,3% eram HPV18. A presença de metástases em linfonodos e os óbitos registrados foram menos frequentes em tumores positivos para HPV, o que sugere melhor prognóstico para esses casos; contudo, as diferenças entre os grupos não foram estatisticamente significantes.

Conclusão

Os resultados obtidos no presente estudo, com respeito à presença de genótipos de alto risco de HPV16 e HPV18, destacam a importância da vacinação para HPV no controle dos carcinomas de cavidade oral e orofaringe.

Palavras-chave
Papillomaviridae; Papillomaviridae 16; Neoplasia de cabeça e pescoço; Epidemiologia

Introduction

Head and neck cancers (HNC), including oral cavity and oropharynx carcinomas, are the sixth most common cancer types in the world, with an annual estimate of 633,000 new cases and 355,000 deaths.11 Chaturvedi AK. Epidemiology and clinical aspects of HPV in head and neck cancers. Head Neck Pathol. 2012;6:16-24. In Brazil, 15,290 HNCs were expected in 2014, with 11,280 cases in men and 4010 in women.22 INCA – Instituto Nacional de Câncer. http://www.inca.gov.br; 2014.
http://www.inca.gov.br...

Oral and oropharynx carcinomas account for more than 80% of the total HNC cases,33 Motta Rda R, Zettler CG, Cambruzzi E, Jotz GP, Berni RB. Ki-67 and p53 correlation prognostic value in squamous cell carcinomas of the oral cavity and tongue. Braz J Otorhinolaryngol. 2009;75:544-9. and squamous cell carcinoma (SCC) is the most common histological type, comprising more than 90% of the cases. The prognosis of these tumors is mostly pessimistic, with a low five-year survival of approximately 58%.44 Montoro JRde MC, Hicz HA, de Souza L, Livingstone D, Melo DH, Tiveron RC, et al. Prognostic factors in squamous cell carcinoma of the oral cavity. Braz J Otorhinolaryngol. 2008;74:861-6.,55 Elango KJ, Suresh A, Erode EM, Subhadradevi L, Ravindran HK, Iyer SK, et al. Role of human papilloma virus in oral tongue squamous cell carcinoma. Asian Pac J Cancer Prev. 2011;12:889-96.

Age, gender, and tumor-node-metastasis (TNM) tumor staging, which includes the extension of the tumor, the presence of lymph node metastases, and distant metastasis, are the main prognostic factors for oral cavity and oropharynx carcinomas.66 Oliveira LR, de Alfredo RS, Sergio Z. Incidence and survival profile of patients with oral squamous cell carcinoma in a Brazilian population. J Bras Patol Med Lab. 2006;42:385-392. In addition, histological grade and the expression of molecular markers (p16, pRb, and Ki-67) allow a better understanding of tumor behavior and evolution.66 Oliveira LR, de Alfredo RS, Sergio Z. Incidence and survival profile of patients with oral squamous cell carcinoma in a Brazilian population. J Bras Patol Med Lab. 2006;42:385-392.88 Saini R, Khim TP, Rahman SA, Ismail M, Tang TH. High-risk human papillomavirus in the oral cavity of women with cervical cancer, and their children. Virol J. 2010;7:131.

The treatment of SCC of the oral cavity and oropharynx is usually accomplished by surgery or radiotherapy, alone or associated, and may also include the use of chemotherapy as an alternative to improve the chances of cure.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4. Studies suggest that for Human papillomavirus (HPV)-positive oral cavity and oropharynx SCC, treatment with surgery and adjuvant radiotherapy is as good as the definitive radiotherapy treatment, with or without chemotherapy.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4.,1010 Hong AM, Dobbins TA, Lee CS, Jones D, Harnett GB, Armstrong BK, et al. Human papillomavirus predicts outcome in oropharyngeal cancer in patients treated primarily with surgery or radiation therapy. Br J Cancer. 2010;103:1510-7.

Smoking and alcohol consumption are considered the main risk factors for oral cavity and oropharynx cancer.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4.1111 Termine N, Panzarella V, Falaschine S, Russo A, Mitranga D, Muzio LL, et al. HPV in oral squamous cell carcinoma vs head and neck squamous cell carcinoma biopsies: a meta-analysis (1988–2007). Ann Oncol. 2008;19:1681-90. However, with the intensification of campaigns against smoking and alcohol, the role of HPV in oral cavity and oropharynx carcinomas has gained prominence in recent years.1111 Termine N, Panzarella V, Falaschine S, Russo A, Mitranga D, Muzio LL, et al. HPV in oral squamous cell carcinoma vs head and neck squamous cell carcinoma biopsies: a meta-analysis (1988–2007). Ann Oncol. 2008;19:1681-90.,1212 Quintero K, Giraldo GA, Uribe ML, Baena A, Lopez C, Alvarez E, et al. Human papillomavirus types in cases of squamous cell carcinoma of head and neck in Colombia. Braz J Otorhinolaryngol. 2013;79:375-81. A growing number of studies support the hypothesis of HPV association with oral cavity and oropharynx carcinomas.1010 Hong AM, Dobbins TA, Lee CS, Jones D, Harnett GB, Armstrong BK, et al. Human papillomavirus predicts outcome in oropharyngeal cancer in patients treated primarily with surgery or radiation therapy. Br J Cancer. 2010;103:1510-7.1414 Muñoz N, Castellsagué X, de González AB, Gissmann L. HPV in the etiology of human cancer. Vaccine. 2011;6:1-10.

HPV is a sexually-transmitted infection; therefore, factors such as early initiation of sexual activity, high number of sex partners, and the practice of unprotected oral sex are included as risk factors for HPV infection in the oral cavity and oropharynx mucosa.1515 Oliveira LR, Ribeiro-Silva A, Ramalho LN, Simões AL, Zucoloto S. HPV infection in Brazilian oral squamous cell carcinoma patients and its correlation with clinicopathological outcomes. Mol Med Rep. 2008;1:123-9. Growing incidence of oral cavity and oropharynx carcinomas associated with HPV in young people has been demonstrated.1414 Muñoz N, Castellsagué X, de González AB, Gissmann L. HPV in the etiology of human cancer. Vaccine. 2011;6:1-10.1616 Zhao D, Xu QG, Chen XM, Fan MW. Human papillomavirus as an independent predictor in oral squamous cell cancer. Int J Oral Sci. 2009;1:119.

The prevalence of HPV in oral cavity and oropharynx SCC is the focus of several studies worldwide; HPV16 is considered the most prevalent and relevant genotype in the epidemiology of these carcinomas.1313 Kleter B, van Doorn LJ, ter Schegget J, Schrauwen L, van Krimpen K, Burger M, et al. Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. Am J Pathol. 1998;153:1731-9.,1717 Lim KP, Hamid S, Lau SH, Teo SH, Cheong SC. HPV infection and the alterations of the pRB pathway in oral carcinogenesis. Oncol Rep. 2007;17:1321-6.2020 Aguiar MTM, de Castro Bosso NC, de Souza Leal CBQ, de Lira CF, Cabral LAO, Silva AMTC, et al. Clinicopathological aspects and prevalence of human papillomavirus in anal cancer. J Coloproctol. 2014;34:76-82.

This study aimed to investigate the prevalence and genotypic distribution of HPV16 and HPV18 in oral cavity and oropharynx carcinomas, as well as their possible association with the clinical characteristics of the tumors.

Methods

Type of study and series

The study was approved by the Research Ethics Committee, under No. 13580613.5.0000.0031/2014. It was a retrospective cross-sectional study that used data collected from medical files and analysis of paraffin blocks containing specimens of oral cavity and oropharynx carcinomas. The selection was first accomplished by analyzing the records of the Pathology Department at the Hospital, and only patients with histological diagnosis of SCC of oral cavity and oropharynx were included. Initially 312 cases of SCC diagnosed in the period from 2005 to 2007 were selected. After eliminating duplicates, 174 cases with available clinical records were selected. Patients who received chemotherapy/radiotherapy before surgery were excluded, resulting in 108 cases. Paraffin blocks from 108 cases were histologically examined, and specimens with exiguous amount of tumors were excluded, resulting in 82 cases that were selected for DNA extraction and HPV DNA detection.

DNA extraction

Genomic DNA was purified from tumor samples fixed in formalin and included in paraffin. The samples were dewaxed in xylene and washed in ethanol according to standardized protocol. DNA was isolated by using the commercial Wizard Kit (Promega). The presence and the integrity of the DNA were verified by the amplification of a 99 base pair (BP) fragment from glyceraldehyde-3-phosphate dehydrogenase (GAPDH), by using polymerase chain reaction (PCR).

HPV DNA detection

HPV DNA was detected by PCR. The set of primers used was SPF 1/2 (short PCR fragment). The SPF 1/2 amplifies a fragment of 65 pb of the L1 region of HPV genome. This set of primers allow the detection of 39 high and low oncogenic risk HPV: 6, 11, 13, 18, 26, 30, 31, 33, 34, 35, 39, 40, 42, 43, 44, 45, 51, 52, 53, 54, 55, 56, 58, 59, 61, 62, 64, 66, 67, 68, 69, 70, 72, MM4, MM7, 73, 74, and MM8. The genotypes detected by SPF 1/2 are those that infect the mucous membrane.1313 Kleter B, van Doorn LJ, ter Schegget J, Schrauwen L, van Krimpen K, Burger M, et al. Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. Am J Pathol. 1998;153:1731-9. PCR with SPF 1/2 primers was carried out in a final reaction volume of 25 µL, adding 2 µL of purified DNA, 2.5 mM/L MgCl2, 2 mM/L of each deoxyribonucleotide (dNTPs), 2.5 µM/L of each oligonucleotide primer, 0.25 U Taq polymerase (Invitrogen, Brazil) and ultra pure water in sufficient quantity for the final volume. Cycling conditions included: preheating for 1 min at 94 ºC, followed by 40 cycles of: 94 ºC for 1 min, 1 min at 45 ºC, and 1 min at 72 ºC, with a final extension of 5 min at 72 ºC. Positive and negative controls were used in each reaction.

Genotyping of HPV16 and 18

HPV16 and HPV18 genotyping was performed by PCR for all HPV-positive tumors. Two sets of primers that amplify part of E7 region of each HPV genome were employed.2121 Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, et al. Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol. 1999;189:12-9. For the HPV16 genome, the amplicon presents 108 pb, and for HPV18 genome amplification, the amplicon presents 104 pb. PCR with HPV16 primers was performed in a final reaction volume of 25 µL, with 2 µL of purified DNA, 2.5 mM/L MgCl2, 2 mM/L of each dNTPs, 2.5 µM/L of each oligonucleotide primer, 0.25 U Taq polymerase (Invitrogen, Brazil), and ultra pure water in sufficient quantity for the final volume. Cycling conditions included: preheating for 1 min at 94 ºC, followed by 40 cycles of: 94 ºC for 1 min, 1 min at 45 ºC, and 1 min at 72 ºC, with a final extension of 5 min at 72 ºC. PCR with HPV18 primers was performed in a final reaction volume of 25 µL, with 5 µL of purified DNA, 2.5 mM/L MgCl2, 2 mM/L of each dNTPs, 2.5 µM/L of each oligonucleotide primer, 1.25 U Taq polymerase, (Invitrogen, Brazil) and ultra pure water in sufficient quantity for the final volume. Cycling conditions included: preheat for 3 min at 94 ºC, followed by 35 cycles of: 1 min at 94 ºC, 1 min at 53 ºC, and 1 min at 72 ºC, with a final extension of 3 min at 72 ºC.

Statistical analysis

All the patients' data were transcribed to Microsoft Excel® spreadsheets. Clinical and histological data of the group, as well as the presence of HPV genome, HPV16, and HPV18 genotypes were analyzed by using Fisher's exact test and the chi-squared test. Values of p ≤ 0.05 were considered statistically significant. Positive and negative controls were used in each reaction.

Results

Clinical and histological characteristics: a group of 82 cases of SCC of oral cavity and oropharynx were selected and evaluated with respect to clinical and histological characteristics (Table 1). Among the patients, 78% were male. Most of the patients (54.9%) were in the age group under 59 years and 62.2% were married. Risk factors, such as smoking (78%) and alcohol consumption (70.8%) were recorded for the majority of the group. Data associated with sexual behavior and orientation, such as the number of sexual partners, age at sexual activity onset, and practice of oral sex were not reported in the medical files.

Table 1
Analysis of the clinical and histological characteristics of the patients with oral cavity and oropharynx carcinomas.

HPV detection and genotyping: HPV DNA was detected in 21 cases (25.6%; 95% confidence interval (CI) 16.9–36.6), of which 33.3% were HPV16 and 14.3% were HPV18. Table 2 describes the clinical characteristics of HPV-positive and negative cases. Among the 21 HPV-positive samples, 47.4% were located in the oral cavity and 52.6% in oropharynx. Considering the clinical staging for HPV-positive tumors, 4.8% were in stages I/II, while 95.2% were in stages III/IV. Lymph node metastases were detected in 42.9% of HPV-positive cases and in 57.4% of HPV-negative cases (p = 0.08). Distant metastases were not detected in HPV-positive cases, while one distant metastasis was described in the HPV-negative cases (p = 0.46). With respect to histological grade, 85.7% of HPV-positive cases showed moderate to high grade of differentiation. A greater number of deaths were registered in the HPV-negative group (39.3%) compared to the HPV-positive group (19.1%), though this difference was not statistically significant (p = 0.11). With respect to HPV genotyping, seven samples (33.3%) were HPV16 positive, while three samples (14.3%) were HPV18 positive. Significant differences between HPV16 and HPV18 tumors were not detected in this study.

Table 2
Analysis of the clinical and histological characteristics of the patients with HPV-positive and HPV-negative oral cavity and oropharynx carcinomas.

Discussion

In this study analyzing 82 cases of SCC of oral cavity and oropharynx diagnosed in a cancer reference center in the central region of Brazil. HPV DNA was detected in 21 cases (25.6%), of which 33.3% were HPV16 and 14.3% were HPV18. These results support the hypothesis that a subgroup of oral cavity and oropharynx carcinomas is HPV related.1212 Quintero K, Giraldo GA, Uribe ML, Baena A, Lopez C, Alvarez E, et al. Human papillomavirus types in cases of squamous cell carcinoma of head and neck in Colombia. Braz J Otorhinolaryngol. 2013;79:375-81.1818 Montaldo C, Mastinu A, Zorco S, Santini N, Pisano E, Piras V, et al. Distribution of human papillomavirus genotypes in Sardinian patients with oral squamous cell carcinoma. Open Virol J. 2010;4:163-8. Significant changes in the epidemiology of mucosal SCCs of the head and neck, with an increasing number of cases related to HPV, have been demonstrated in the last decade.11 Chaturvedi AK. Epidemiology and clinical aspects of HPV in head and neck cancers. Head Neck Pathol. 2012;6:16-24.2222 Westra WH. The changing face of head and neck cancer in the 21st century: the impact of HPV on the epidemiology and pathology of oral cancer. Head Neck Pathol. 2009;3:78-81. In addition to tobacco and alcohol consumption, HPV appears as an important risk factor for oral cavity and oropharynx SCC development. The prevalence of HPV DNA in oropharyngeal cancer (OC) varies in different studies, and up to 84% of cases have been associated to HPV infection.1919 Krüger M, Pabst AM, Walter C, Sagheb K, Günther C, Blatt S, et al. The prevalence of human papilloma virus (HPV) infections in oral squamous cell carcinomas: a retrospective analysis of 88 patients and literature overview. J Craniomaxillofac Surg. 2014;42:1506-14.

The association between HPV infection with oral cavity and oropharynx SCC emphasizes the importance of introducing specific molecular tests into oral cancer prevention practices, in order to evaluate the presence of the virus and the possibility of expanding anti-HPV vaccine in the male population.2323 Vacinação contra HPV supera meta de 80%. Portal da Saúde – Ministério da Saúde. N.p., n.d. Available from: http://www.saude.gov.br [cited 2.12.14].
http://www.saude.gov.br...
,2424 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. Evaluation of a combined triple method to detect causative HPV in oral and oropharyngeal squamous cell carcinomas: p16 immunohistochemistry, consensus PCR HPV-DNA, and in situ hybridization. Infect Agent Cancer. 2012;7:4.

In the present study, 54.9% of the patients were younger than 59 years. Several studies describe the age range of the patients with carcinoma of oral cavity and oropharynx as similar to the value described in the present study.2525 Wittekindt C, Wagner S, Mayer CS, Klussmann JP. Basics of tumor development and importance of human papilloma virus (HPV) for head and neck cancer. GMS Curr Top Otorhinolaryngol Head Neck Surg. 2012;11.,2626 Angadi PV, Savitha JK, Rao SS, Sivaranjini Y. Oral field cancerization: current evidence and future perspectives. Oral Maxillofac Surg. 2012;16:171-80. In a study conducted in Italy, the median age was 65.6 years.1818 Montaldo C, Mastinu A, Zorco S, Santini N, Pisano E, Piras V, et al. Distribution of human papillomavirus genotypes in Sardinian patients with oral squamous cell carcinoma. Open Virol J. 2010;4:163-8. The accumulation of exposure to various risk factors, such as lifelong smoking and alcoholism, contributes to the higher prevalence of these tumors in more advanced ages.66 Oliveira LR, de Alfredo RS, Sergio Z. Incidence and survival profile of patients with oral squamous cell carcinoma in a Brazilian population. J Bras Patol Med Lab. 2006;42:385-392. Studies demonstrated that HPV-associated HNC, including oropharyngeal and oral cavity SCCs, have recently risen dramatically in men under 50 years old.2727 Nordfors C, Vlastos A, Du J, Ährlund-Richter A, Tertipis N, Grün N, et al. Human papillomavirus prevalence is high in oral samples of patients with tonsillar and base of tongue cancer. Oral Oncol. 2014;50:491-7. In a period of 20 years, the relative prevalence of HPV-positive oropharyngeal squamous cancer cell (OSCC) went from less than 20% to more than 70% in the United States and some European countries.2828 Gillison ML, Koch WM, Capone RB, Spafford M, Westra WH, Wu L, et al. Evidence for a causal association between human papillomavirus and a subset of head and neck cancers. J Natl Cancer Inst. 2000;92:709-20.3030 Sathish N, Wang X, Yuan Y. Human papillomavirus (HPV)-associated oral cancers and treatment strategies. J Dent Res. 2014;93:29S-36S. In the HPV-positive patients evaluated in the present series, 66.7% of the cases were under the average age (59 years). Most cases of carcinoma of the oral cavity and oropharynx were observed in males (78.0). Various authors have previously reported this information; however, in different countries of Europe, an increasing tendency in the incidence of oropharynx carcinomas has been noticed in females.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4.2929 Snietura M, Piglowski W, Jaworska M, Mucha-Malecka A, Wozniak G, Lange D, et al. Impact of HPV infection on the clinical outcome of p-CAIR trial in head and neck cancer. Eur Arch Otorhinolaryngol. 2011;268:721-6.

Smoking and alcohol consumption are described as the main risk factors for carcinomas of the oral cavity and oropharynx.1111 Termine N, Panzarella V, Falaschine S, Russo A, Mitranga D, Muzio LL, et al. HPV in oral squamous cell carcinoma vs head and neck squamous cell carcinoma biopsies: a meta-analysis (1988–2007). Ann Oncol. 2008;19:1681-90.3131 Chaturvedi AK, Engels EA, Pfeiffer RM, Hernandez BY, Xiao W, Kim E, et al. Human papillomavirus and rising oropharyngeal cancer incidence in the United States. J Clin Oncol. 2011;29:4294-301. The present study confirmed the high frequency of smokers (78.0%) and alcohol drinkers (70.8%) in the group. Although not fully considered as a prognostic factor, carcinomas of the oral cavity and oropharynx associated with smoking and alcoholism tend to be more aggressive.11 Chaturvedi AK. Epidemiology and clinical aspects of HPV in head and neck cancers. Head Neck Pathol. 2012;6:16-24. Tobacco and alcohol can induce SCC of the oral cavity and oropharynx by aggression on extensive areas, leading to a phenomenon known as field cancerization, which is characterized by molecular changes in the reserve cells, leading to the formation of epithelial field changes.2626 Angadi PV, Savitha JK, Rao SS, Sivaranjini Y. Oral field cancerization: current evidence and future perspectives. Oral Maxillofac Surg. 2012;16:171-80. This field undergoes expansion and extends over the surface of the mucous membrane, increasing the possibility of formation of a new carcinoma.3232 Huang S-F, Li H-F, Liao C-T, Wang H-M, Chen I-H, Chang J-C, et al. Association of HPV infections with second primary tumors in early staged oral cavity cancer. Oral Dis. 2012;18:809-15.,3333 Chung CH, Ashley B, Gypsyamber DS. Epidemiology of oral human papillomavirus infection. Oral Oncol. 2014;50:364-9.

The profile of patients with carcinoma of the oral cavity and oropharynx associated with HPV tends to be characterized by a group of younger patients, aged less than 60 years, non-smokers or light smokers, non-drinkers, and with more promiscuous sexual behavior; however, these data are controversial.3434 Upile T, Jerjes W, Al-Khawalde M, Radhi H, Sudhoff H. Oral sex, cancer and death: sexually transmitted cancers. Head Neck Oncol. 2012;4:31.,3535 D'Souza G, Cullen K, Bowie J, Thorpe R, Fakhry C. Differences in oral sexual behaviors by gender, age, and race explain observed differences in prevalence of oral human papillomavirus infection. PLOS ONE. 2014;9:e86023. One important limitation of the present study is the lack of data related to the patients' sexual behavior. Since it was a retrospective study, such data were not available in the medical files. Although a higher proportion of patients were described under 59 years in the HPV-positive group (66.7%) compared to the HPV-negative group (50.8%), these differences were not statistically significant. Similar results with respect to age group were described in two studies developed in the United States.1313 Kleter B, van Doorn LJ, ter Schegget J, Schrauwen L, van Krimpen K, Burger M, et al. Novel short-fragment PCR assay for highly sensitive broad-spectrum detection of anogenital human papillomaviruses. Am J Pathol. 1998;153:1731-9.,1414 Muñoz N, Castellsagué X, de González AB, Gissmann L. HPV in the etiology of human cancer. Vaccine. 2011;6:1-10. In the present study, the average age for HPV-positive patients was lower, 53 years, compared to the HPV-negative group, 60 years. A study conducted in Colombia confirmed a lower average age for HPV-positive patients with oral cavity and oropharynx carcinomas.1212 Quintero K, Giraldo GA, Uribe ML, Baena A, Lopez C, Alvarez E, et al. Human papillomavirus types in cases of squamous cell carcinoma of head and neck in Colombia. Braz J Otorhinolaryngol. 2013;79:375-81. Significant associations between the presence of HPV and the absence of habits like smoking and alcohol consumption were not noticed in the present study, corroborating with various studies.66 Oliveira LR, de Alfredo RS, Sergio Z. Incidence and survival profile of patients with oral squamous cell carcinoma in a Brazilian population. J Bras Patol Med Lab. 2006;42:385-392.1818 Montaldo C, Mastinu A, Zorco S, Santini N, Pisano E, Piras V, et al. Distribution of human papillomavirus genotypes in Sardinian patients with oral squamous cell carcinoma. Open Virol J. 2010;4:163-8.

The prognosis of oral cavity and oropharynx carcinomas is uncertain and difficult to predict.44 Montoro JRde MC, Hicz HA, de Souza L, Livingstone D, Melo DH, Tiveron RC, et al. Prognostic factors in squamous cell carcinoma of the oral cavity. Braz J Otorhinolaryngol. 2008;74:861-6. The identification of factors that may help choosing the best treatment, predicting the evolution of the tumor as well as patient's survival, is of evident clinical importance. HPV-positive oral cavity and oropharynx SCC seem to present a better prognosis when treated with surgery and adjuvant radiotherapy or radiation therapy with or without final chemotherapy.1010 Hong AM, Dobbins TA, Lee CS, Jones D, Harnett GB, Armstrong BK, et al. Human papillomavirus predicts outcome in oropharyngeal cancer in patients treated primarily with surgery or radiation therapy. Br J Cancer. 2010;103:1510-7. Limitations on data collection in the present study prevented a survival analysis associated with the therapy used. A systematic review highlights the importance of evaluating patients with HPV-related oropharyngeal carcinomas, in order to predict the best therapy, since these virus-associated carcinomas show distinct molecular characteristics compared to HPV-negative ones.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4. However, currently, only a few studies describe a direct association between treatment, the presence of the HPV genome, and survival.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4.,1010 Hong AM, Dobbins TA, Lee CS, Jones D, Harnett GB, Armstrong BK, et al. Human papillomavirus predicts outcome in oropharyngeal cancer in patients treated primarily with surgery or radiation therapy. Br J Cancer. 2010;103:1510-7.

A better prognosis for HPV-positive oral cavity and oropharynx carcinomas has been described in a few studies, including well-differentiated tumors, with less risk of lymph node metastasis and distant metastasis.99 Pannone G, Santoro A, Papagerakis S, Lo Muzio L, De Rosa G, Bufo P. The role of human papillomavirus in the pathogenesis of head & neck squamous cell carcinoma: an overview. Infect Agent Cancer. 2011;6:4.3636 Pytynia KB, Kristina RD, Erich MS. Epidemiology of HPV-associated oropharyngeal cancer. Oral Oncol. 2014;50:380-6. In the present study, these characteristics were observed; however, the difference between the groups was not significant. Distant metastases were not described in the HPV-positive group, although only one case of HPV-negative tumors presented with distant metastases. Concerning the presence of lymph node metastasis, a lower rate of lymph node metastasis was observed in the HPV-positive tumors (42.9% vs. 57.4%), suggesting a better prognosis for these tumors; however, such differences were not statistically significant.

HPV16 and HPV18 are described as the most prevalent genotypes in oral cavity and oropharynx carcinomas, and their association with these carcinomas seems to be relevant.1414 Muñoz N, Castellsagué X, de González AB, Gissmann L. HPV in the etiology of human cancer. Vaccine. 2011;6:1-10. In the present study, of the 21 HPV-positive samples, HPV16 was described in 33.3% and HPV18 in 14.3% of the tumors. To describe the prevalence of HPV16 and 18 in oral cavity and oropharynx carcinomas is important in order to predict the impact of vaccination on these tumors, since both genotypes are the main targets for the bivalent (16/18) and quadrivalent vaccines (11/16/18/6).1515 Oliveira LR, Ribeiro-Silva A, Ramalho LN, Simões AL, Zucoloto S. HPV infection in Brazilian oral squamous cell carcinoma patients and its correlation with clinicopathological outcomes. Mol Med Rep. 2008;1:123-9.2727 Nordfors C, Vlastos A, Du J, Ährlund-Richter A, Tertipis N, Grün N, et al. Human papillomavirus prevalence is high in oral samples of patients with tonsillar and base of tongue cancer. Oral Oncol. 2014;50:491-7. In the present study, no significant differences were described regarding the staging and death of the patients when comparing HPV16 and 18 positive and negative cases.

Recent reports highlight that vaccination campaigns are an efficient solution for the control of the HPV-associated carcinomas.1515 Oliveira LR, Ribeiro-Silva A, Ramalho LN, Simões AL, Zucoloto S. HPV infection in Brazilian oral squamous cell carcinoma patients and its correlation with clinicopathological outcomes. Mol Med Rep. 2008;1:123-9.3737 Ringström E, Peters E, Hasegawa M, Posner M, Liu M, Kelsey KT. Human papillomavirus type 16 and squamous cell carcinoma of the head and neck. Clin Cancer Res. 2002;8:3187-92. Regarding this connection and considering the high prevalence of HPV in oral cavity and oropharynx carcinomas in males, it is extremely important that this campaign might also be extended to the male group.

In the present study, the presence of HPV was detected in 25.6% of the cases, including types 16 and 18, leading to the conclusion that HPV is also associated with the development of oral cavity and oropharyngeal carcinomas. This study presented an important limitation with respect to clinical and behavioral data of the patients. In the medical files, such data were scarce, including sexual behavior, oral hygiene, history of sexually transmitted diseases, and patient follow-up, among others. These difficulties are inherent to retrospective studies, since the researchers rely only on the information present in medical records that can often be lost, incomplete, or unclear.2020 Aguiar MTM, de Castro Bosso NC, de Souza Leal CBQ, de Lira CF, Cabral LAO, Silva AMTC, et al. Clinicopathological aspects and prevalence of human papillomavirus in anal cancer. J Coloproctol. 2014;34:76-82.3030 Sathish N, Wang X, Yuan Y. Human papillomavirus (HPV)-associated oral cancers and treatment strategies. J Dent Res. 2014;93:29S-36S.

The authors suggest a prospective study, with more efficient data collection and patient follow-up, allowing a more accurate and complete survey, with a larger number of cases from different oncology centers in the country, in order to improve the knowledge of HPV carcinogenesis in oral cavity and oropharynx carcinomas.

Conclusions

This study confirms the higher prevalence of HPV DNA in oral cavity and oropharynx carcinomas, especially in males (78%), with an average age of 58 years, and a high frequency of smokers and alcohol drinkers.

The prevalence of HPV DNA genome in the samples analyzed was 25.6%, and among the positive cases, 33.3% were HPV16 and 14.3% were HPV18, highlighting the association of high-risk HPV in oral cavity and oropharynx carcinogenesis.

  • Please cite this article as: Petito G, Carneiro MA, Santos SH, Silva AM, Alencar RC, Gontijo AP. Human papillomavirus in oral cavity and oropharynx carcinomas in the central region of Brazil. Braz J Otorhinolaryngol. 2017;83:38-44.
  • Peer Review under the responsibility of Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial.

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Publication Dates

  • Publication in this collection
    Jan-Feb 2017

History

  • Received
    16 Sept 2015
  • Accepted
    10 Jan 2016
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