口腔鳞状细胞癌中人乳头瘤病毒感染的检测

目的检测人乳头瘤病毒(HPV)在口腔鳞状细胞癌中感染及其与口腔癌发生的相关性.方法采用PCR方法,检测30例口腔鳞状细胞癌及5例正常口腔黏膜中HPV 16和18感染.结果 7/30例标本中检测出HPV 16(23.3%);10/30例标本中检测出HPV 18(33.3%).3/30例标本中存在HPV 16和18共感染(10%).5例正常口腔黏膜中未发现HPV 16和HPV 18感染.在检测HPV 16时发现一条新片断(186 nt),经测序此序列与人5号染色体的核酸序列65-175存在99%同源.结论 HPV 16和18可能与口腔鳞状细胞癌的发生密切相关.     

Low prevalence of human papillomavirus infection in the healthy oral mucosa of a Northern Italian population

J Oral Pathol Med (2012) 41 : 16–20 Background: Oral cancer is the sixth most common malignancy in developed countries, representing almost 3% of malignant tumors. Tobacco use and alcohol consumption are well‐established risk factors. However, the observation that most patients with oral cancer have not been exposed to these risk factors suggests that additional causes may promote oral carcinogenesis. A link has been suggested between human papillomavirus (HPV) and oral cavity cancer but the significance of HPV contribution to oral carcinogenesis as well as the prevalence of HPV infection in normal oral cavity mucosa remains debated. Methods: In this study, the prevalence of oral HPV infection was evaluated in 81 randomly selected Northern Italian subjects with clinically normal oral mucosa using a nested PCR on DNA extracted by oral smears. Results and conclusions: No HPV‐related lesions were detectable in any of the smears analyzed by cytological approach. nPCR identified HPV DNA in only one (1.2%) of the specimens obtained from clinically healthy oral mucosa and subsequent characterization assigned the positive case to HPV type 90. These data suggest that the incidence of HPV infection in the healthy population might be very low and that other risk factors are likely responsible to promote oral carcinogenesis.     

口腔鳞状细胞癌高危型人乳头状瘤病毒感染的检测

目的 观察口腔鳞癌及口腔粘膜高危型人乳头状瘤病毒（HPV）感染情况,探讨HPV感染与临床及病理资料的关系。方法 用多聚酶链反应（PCR）检测73例口腔鳞癌及40例正常口腔粘膜石蜡包埋组织中HPV16、HPV18的DNA。统计分析其与临床及病理资料的关系。结果 口腔鳞癌HPV16／HPV18 DNA阳性率为74％（54／73）,正常口腔粘膜为55％（22／40）。口腔鳞癌与正常口腔粘膜HPV阳性率存在显著性差异（P＝0．040）。统计分析显示：HPV感染与患者的性别、年龄有关,与其它因素（肋瘤发生部位、嗜烟酒情况、肿瘤病理分级、临床分期）无关。结论 高危型HPV感染与口腔鳞癌的发生有关。口腔粘膜中HPV感染普遍存在,提示HPV在口腔肿瘤的发生中并非独立发挥作用。     

Human papillomavirus type 16 and 18 DNA in oral squamous cell carcinoma and normal mucosa

The aim of this study was to ascertain the prevalence of HPV 16/18 DNA in oral squamous cell carcinoma (OSCC) vs. normal oral mucosa, and to correlate the virologic data with other factors obtained from the patients' records. One hundred and thirteen paraffin embedded tissue samples (73 OSCC and 40 normal oral mucosa) were studied using HPV type specific primer-mediated polymerase chain reaction (PCR). Seventy-four per cent (54/73) of OSCC and 55% (22/44) of normal oral mucosa were positive for HPV 16/18 DNA. Statistical analysis indicated there was a significant difference between HPV16/18 positive OSCC vs. normal oral mucosa (P=0.040), and that age (<60 years) and gender (male) were correlated with the presence of HPV16/18 in the tumour. No significant association was found between the presence of HPV and other risk factors, including tobacco use, alcohol use, tumour location, histologic grade or TNM staging. We found a significant association of HPV16/18 with oral squamous cell carcinoma. Also, HPV16/18 is a co-factor in oral carcinogenesis, particularly in male patients and patients under the sixth decade. In addition, we found that HPV infection is a common event in the normal oral mucosa.     

High risk human papillomavirus in oral squamous carcinoma: evidence of risk factors in a Venezuelan rural population. Preliminary report

In a search for the presence of human papillomavirus (HPV) and some etiologic cofactors in oral squamous cell carcinoma (OSCC), 50 women diagnosed as OSCC were analyzed by a multiplex polymerase chain reaction (PCR) assay specific for HPV types 6, 11, 16, and 18. This study revealed that 60% (30/50) of the OSCC patients were positive for HPV-DNA sequences. This group was analyzed according to smoking, alcohol consumption, number of pregnancies, poor oral health and low social economic status. The current results indicate an increased incidence of HPV malignant types in the oral cavity in women with OSCC. Also, they support a multifactorial model of oral cancer causation.     

口腔鳞癌中HPV16、18型感染和p53蛋白表达的检测研究

目的探讨人类乳头状瘤病毒(human papillomavirus, HPV)感染、p53蛋白表达在口腔鳞状细胞癌发生、发展中的作用与相互关系.方法采用聚合酶链反应(PCR)检测HPV16、18型DNA;采用免疫组织化学LSAB法检测p53蛋白产物在细胞核中的表达.结果口腔鳞癌组中HPV16、18 DNA总阳性率为48.89%(22/45),p53蛋白表达阳性率为62.22%(28/45).HPV16、18型感染组及p53蛋白过度表达组的平均生存期、生存率均低于无HPV感染组(P>0.05)及无p53过度表达组(P<0.05).结论 HPV16、18型感染、p53基因突变与口腔鳞癌的发生密切相关.口腔鳞癌患者的预后在一定程度上与HPV感染、p53表达状况有关.     

The role of human papillomavirus in oral squamous cell carcinoma: myth and reality

Introduction

As the traditional risk factors for oral squamous cell carcinoma, especially tobacco, decline, new potential causative agents become the focus of research. Since the discovery of human papillomavirus (HPV) and its importance in carcinogenesis in cervical cancer, a lot of research has been undertaken to define its role in different types of cancer. In the present study, we evaluate the role of high-risk HPV types in initiation and progression of oral squamous cell carcinoma (OSCC) using a systematic review of the current literature.

Material and methods

A literature research with the search term “HPV oral squamous cell carcinoma” was performed via PubMed. Results were screened systematically for relevance and classified into the following categories: molecular biology, genetics, clinical aspects, and prevalence. Articles were then further analyzed to assess quality.

Results

The literature research led to 527 results, with an overall HPV prevalence of 30.1 % in OSCCs. The most frequently identified subtypes were HPV-16 and HPV-18 (25.4 and 18.1 %, respectively). Prognostic relevance of HPV was discussed controversially. HPV detection via polymerase chain reaction is the most established method today. Molecular changes according to carcinogenic pathways described for cervix carcinoma were not routinely found in OSCC. In general, no definite role of high-risk HPV is currently deducible from the literature.

Conclusions

High-risk subtypes 16 and 18 are present in the genome in approximately one third of OSCC. Its role as a causative agent is less clear than the role in oropharyngeal tumors. The infection might not be the cause of carcinogenesis in a significant number of patients but may become proportionally more important with the decrease of the classical risk factors of tobacco and alcohol.     

HPV in oral squamous cell carcinomas of a Brazilian population: amplification by PCR

Human Papilomaviruses (HPV) are a group of viruses associated with benign and malignant lesions of cutaneous and mucosal epithelia. Some "high risk" HPV types, especially HPV 16 and 18, are strongly correlated with cervical and anogenital cancers and are also related to the genesis of oral squamous cell carcinomas (OSCC). The aim of this work was to investigate the incidence of HPV infection in 40 paraffin-embedded or fresh specimens of OSCC, using PCR amplification of the viral DNA. Literature based primers (GP5+/GP6+) were used in order to amplify HPV DNA from the L1 gene, present in more than 22 types of HPV. A condyloma case with HPV 16 and 18 detected by in situ hybridization was used as a positive control. Amplification of HPV was observed only in the positive control. No squamous cell carcinoma cases showed DNA viral amplification. Absence of HPV DNA amplification by PCR in the analyzed specimens of OSCCs suggests that this virus not always plays a role in the carcinogenesis process. Discrepancy with some studies found in the literature may be related to methodology or population differences.     

Association of HPV infections with second primary tumors in early-staged oral cavity cancer

Oral Diseases (2012) 18 , 809–815 Objective: The infection of human papilloma virus (HPV) has been reported in head and neck cancer; however, the clinical significance of HPV infection on the pathogenesis of oral cavity squamous cell carcinoma (OSCC) is still uncertain. Materials and Methods: The study recruited 103 patients with pathological early‐stage OSCC between March 1997 and December 2003 from Chang Gung Memorial Hospital, Taiwan. Tumor specimens were HPV‐genotyped by the EasychipVR HPV Blot method. Clinical association study was performed by using chi‐square, Kaplan–Meier, and logrank tests. Results: Thirty‐one patients (30.1%) were positive for HPV infection. The most frequent HPV types were types 16 (16 patients, 51.6%) and 18 (seven patients, 22.6%). HPV infection was not associated with tumor aggressiveness (pathological tumor stage or differentiation status), risk exposure (alcohol, cigarette, or areca quid chewing habit), or the treatment outcome (disease‐free survival or overall survival). However, infection with HPV‐18 was associated with the occurrence of a second primary cancers (P = 0.033), indicating the infection of HPV in OSCC enhances the susceptibility of developing secondary malignancy. Conclusions: There are 30% of the patients with OSCC infected with HPV, with most high‐risk types. HPV‐18 infection may enhance the susceptibility of second primary tumors. Large scale of validation study will be needed to confirm this result.     

Oral cancer; the evidence for sexual transmission

The incidence of oral cancer amongst young adults is increasing in many European and high incidence countries. Most oral cancer is aetiologically linked to the use of tobacco and/or alcohol but nearly two decades ago, we produced the first evidence for the presence of viral nucleic acids in oral squamous cell carcinoma (OSCC) tissues, hypothesising that there may be a viral involvement in at least some OSCC. Subsequently, human papillomaviruses (HPV) in particular have been implicated in OSCC. Antibody responses to HPV are seen and HPV-DNA detected in tumours by us and many others, the virus being mainly HPV-16, the genotype associated with ano-genital cancer. Recent studies have indicated that HPV may be aetiologically important particularly in some types of oropharyngeal cancer, at least in tonsillar carcinogenesis, and may represent an alternative pathway in carcinogenesis to the established factors of tobacco and alcohol. Studies of patients with OSCC have suggested possible sexual transmission of HPV.     

PCR－RDB技术检测中国籍OSCC及OLK患者中的HPV

目的：研究口腔鳞癌（OSCC）及口腔白斑（OLK）中国患者中的人乳头瘤病毒（Human Papillomavirus, HPV）感染率。方法：提取35例OSCC及35例OLK患者病损组织DNA,通过聚合酶链式反应结合反向点杂交技术（Poly－merase Chain Reaction-Reverse Dot Blot Hybridization, PCR-RDB）检测23型HPV DNA,包括HPV6,11,16,18,31,33,35,39,42,43,45,51,52,53,56,58,59,66,68,73,81,82和83。结果：35例OSCC及35例OLK患者中HPV DNA均为阴性。结论：HPV感染在中国的OSCC及OLK患者中非常见分子事件。     

口腔鳞状细胞癌及白斑人乳头瘤病毒16型感染的临床研究

目的:探讨口腔鳞状细胞癌(OSCC)和白斑(LK)患者人乳头瘤病毒16型(HPV-16)感染和临床病理参数之间的相关性。方法:收集30例OSCC和5例LK患者的组织切片,所有病例均经病理学确诊,提取组织DNA,通过巢式PCR检测HPV-16感染情况。结果:HPV-16与TNM分级之间未见相关性,60岁以下女性患者的HPV-16感染率高于60岁以上女性患者(P=0.024),而男性患者未见HPV-16感染和年龄存在相关性(P=0.649)。结论:60岁以下女性OSCC及LK患者中出现较高的HPV-16感染率,提示这可能是新的致癌危险因子。     

Oral squamous cell carcinoma in north-eastern Hungary. II. Etiological factors

The purpose of this study is to determine the possible etiological factors of oral squamous cell carcinoma (OSCC) in North-Eastern Hungary. The medical records of 119 randomly selected patients with OSCC admitted to the Department of Maxillofacial Surgery of the Faculty of Dentistry, University of Debrecen were reviewed. The following risk factors were investigated: tobacco and alcohol consumption, dental status, rural vs. urban residence, and high risk HPV infection. The presence of HPV DNA has been evaluated by polymerase chain reaction from the tissue samples. Results were correlated with clinical data. At the time of diagnosis 65.5 percent of the patients were smokers. Under the age of 45 the rate was 86.4%. Smoking significantly correlated with younger age, male gender, advanced clinical stages and alcohol consumption. The majority of the patients (75.5%) consumed alcohol, 41.1% regularly over the acceptable range. Drinking habit significantly correlated with younger age, male gender and tumor site (gingiva, retromolar region, tongue). HR-HPV types were detected in 42.8% of samples tested. HPV DNA presence was not related to gender, clinical stage, histological grade or other risk factors. Authors found weak correlation between HR-HPV positivity, younger patient age and better 5-year survival rate. The dental status was acceptable only in 12.6 percent of the cases. There was a correlation between dental status and age, smoking and drinking habits. No significant urban-rural differences were found. In the study population the most important risk factor for developing oral cancer is tobacco smoking followed by alcohol consumption. Avoidance of tobacco smoking and a reduced amount of alcohol, together with healthy nutrition and regular dental care should be emphasized.     

Human papillomavirus as a risk factor for oral squamous cell carcinoma: a meta-analysis, 1982-1997

OBJECTIVE: Human papillomavirus (HPV) infection is a significant risk factor for uterine cervical carcinoma. However, the role of HPV infection in oral squamous cell carcinoma (OSCC) is less well defined. To determine the significance of the relationship of this virus in the progressive development of oral cancer, we estimated the risk of HPV detection in normal oral mucosa, precancerous oral tissue, and oral carcinoma using meta-analysis. STUDY DESIGN: Case reports and clinical series published in English-language journals were retrieved by searching MEDLINE (January 1980-August 1998). Review articles were also examined to identify additional studies. Studies that used biochemical, immunologic, microscopic, or molecular analyses to detect HPV in tissue or cells derived from normal oral mucosa (n = 25), benign leukoplakia (n = 21), intraepithelial neoplasia (ie, dysplasia and carcinoma in situ; n = 27), and oral cancer (n = 94) were included in the meta-analysis. Information on sample size, age, sex, method of tissue preservation (ie, fresh, frozen, paraffin-embedded), assay, primer amplification region (early, late), high-risk versus low-risk genotype, and use of tobacco or alcohol was abstracted by one author (C.S.M.). RESULTS: Data from 94 reports that analyzed 4680 samples were included in the meta-analysis. Analyses made by means of a random-effects model with and without adjustments for assay sensitivity showed increased probability of HPV detection in tissue with precancerous and cancerous features compared with normal mucosa. The likelihood of detecting HPV in normal oral mucosa (10.0%; 95% confidence interval [CI], 6.1%-14.6%) was significantly less than of detecting benign leukoplakia (22.2%; 95% CI, 15.7%-29.9%), intraepithelial neoplasia (26.2%; 95% CI, 19.6%-33.6%), verrucous carcinoma (29.5%; 95% CI, 23%-36.8%), and OSCC (46.5%; 95% CI, 37.6%-55.5%). Adjustment of findings for differences in assay sensitivity indicated that these estimates may be conservative. Overall, HPV was between 2 and 3 times more likely to be detected in precancerous oral mucosa and 4.7 times more likely to be detected in oral carcinoma than in normal mucosa. The pooled odds ratio for the subset of studies directly comparing the prevalence of HPV in normal mucosa and OSCC was 5.37, confirming the trend observed in the overall sample. The probability of detecting high-risk HPVs in OSCCs was 2.8 times greater than that of low-risk HPVs. CONCLUSION: This meta-analysis indicates that HPV is detected with increased frequency in oral dysplastic and carcinomatous epithelium in comparison with normal oral mucosa. The findings provide further quantitative evidence that oral infection with HPV, particularly with high-risk genotypes, is a significant independent risk factor for OSCC.     

HPV and other risk factors of oral cavity/oropharyngeal cancer in the Czech Republic

OBJECTIVE: An association between high-risk human papillomavirus (HR HPV) infection and a risk of development of a subgroup of head and neck cancers has been proposed recently. The main risk factors of oral and oropharyngal cancer observed in our population are smoking and alcohol consumption. The incidence of oral/oropharyngeal tumours in the Czech Republic is relatively high and there are no data available about the prevalence of HPV DNA presence in these tumours. MATERIALS AND METHODS: Eighty patients with a primary oropharyngeal cancer were enrolled. The presence of HPV DNA has been evaluated by polymerase chain reaction in 68 cases from which the tumour tissue and demographical and clinical data were available. The typing of HPV was performed by nucleotide DNA sequencing. RESULTS: The HPV DNA was detected in 51.5% of samples tested. Among the HPV DNA positive tumours, 80% contained HPV16. In the analysed group there were 54 men and 14 women. The prevalence of HPV DNA was lower in oral (25%) than in oropharyngeal (57%) tumours, and higher in never smokers (100%) and never drinkers (68.8%). HPV DNA presence was not related to gender, age, number of lifetime sexual partners or practice of oral-genital sex, size of tumour or presence of regional metastases. CONCLUSIONS: The difference in the prevalence of HPV DNA positive tumours between cases of oral cavity and oropharyngeal carcinoma exposed and not exposed to tobacco or alcohol support the theory that HPV DNA positive tumours form an aetiologically distinct subgroup of head and neck tumours.     

正常口腔黏膜和口腔鳞癌中人乳头瘤病毒感染率的研究进展

黏膜高危型HPV-16、HPV-18感染是宫颈癌的主要致病因素,且与口腔鳞癌的发生密切相关,但目前对口腔鳞癌中HPV的感染率和亚型的分布尚不十分清楚。作者系统查阅了目前已经发表的有关HPV在正常口腔黏膜或口腔鳞癌中感染率和亚型分布的文献资料,对正常口腔黏膜、口腔鳞癌中人乳头瘤病毒的感染率的研究进展进行了综述。     

Acetowhite staining and its significance in diagnosis of oral mucosal lesions in women with genital HPV infections

White staining of genital mucosa after acetic acid application (3-5%) (acetowhiteness) has been commonly regarded as a sign of HPV infection, and acetic acid application is widely used as a routine diagnostic means to screen the HPV infections. However, the sensitivity and specificity of acetowhite staining to detect HPV infections has not been properly studied. The usefulness of acetic acid application in detecting oral HPV infections is not established. In this communication, the oral mucosal changes after acetic acid application were systematically recorded in 315 women prospectively followed-up for genital HPV infections, with special reference to smoking, alcohol consumption, histology, cytology and presence of HPV DNA. Strong and weak acetowhite staining were found in 2% and 38% of the patients, respectively. The mean age of these patients was 30.8 yr in contrast to 37.3 yr of the patients without acetowhite lesions, the difference being significant. In most cases, the staining was widely distributed over the buccal mucosa. Acetowhiteness on oral mucosa was seen significantly more frequently in smokers, but the staining did not show any correlation with alcohol consumption, histologic and cytologic findings, presence of HPV DNA or with glycogen content. The specificity of acetowhite staining to detect HPV DNA was 50%. Vacuolized cells were found significantly more often in PAS-positive biopsies. The mean day of the menstrual cycle of the acetic acid--positive and -negative patients at examination was 14.9 and 11.9, respectively. The results suggest that positive acetic acid staining in oral cavity should not be regarded as a diagnostic criteria for HPV infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Age-dependence of human papillomavirus DNA presence in oral squamous cell carcinomas

The aetiology of oral cancer is thought to be multifactorial. Apart from the two known major risk factors (tobacco and alcohol), a viral aetiology has been proposed, with special reference to human papillomavirus (HPV).35 cases of oral squamous cell carcinoma (OSCC), seen at the Departments of Oral & Maxillofacial Surgery and Oral Pathology and Otolaryngology of the Free University of Amsterdam, were analysed as well as 12 biopsies of clinically and histologically normal gingival mucosa collected from healthy individuals after tooth extractions, using the polymerase chain reaction (PCR) and two different sets of primers that are able to detect a broad spectrum of HPV types.An overall HPV positivity of 54.3% in OSCC was found, the majority of positive cases (78.9%) harbouring HPV type 16. In contrast, no positivity for HPV was detected in the clinically normal oral mucosal samples analysed. Furthermore, a significant association between HPV presence and age was found: patients older than 60 years showed a lower prevalence of the virus (29.4%) compared with patients below this age (77.8%) (P < 0.05). The results from the present study suggest an association between HPV and OSCC, particularly in patients under the seventh decade.     

Risk of oral cancer associated with human papillomavirus infection, betel quid chewing, and cigarette smoking in Taiwan — an integrated molecular and epidemiological study of 58 cases

BACKGROUND: The association between oral squamous cell carcinoma (OSCC) and human papillomavirus (HPV) 6, 11, 16 and 18 is uncertain. Past reports varied in the methodology and results. We conducted this study using in situ PCR in situ hybridization (ISH) assay which was considered as the most sensitive method for detection of viral DNA. We undertook an epidemiologic survey about the history of betel quid chewing and cigarette smoking, since these habits are common in Taiwan. METHODS: In situ PCR ISH was performed on the tumor specimens from 29 patients with OSCC and the oral mucosal specimens from 29 patients without OSCC. Their betel quid chewing and cigarette smoking histories were also reviewed. RESULTS: HPV16, HPV18, betel quid chewing and cigarette smoking were statistically significant risk factors in univariate analysis. HPV6 and 11 were not. Multivariate analysis showed that HPV16 infection (adjusted Odds ratio = 11.20) and betel quid chewing (adjusted Odds ratio = 17.06) remained to be independent factors for OSCC. CONCLUSIONS: Our results showed that HPV16 and betel quid chewing were two major risk factors for OSCC in Taiwan, indicating that they act through different mechanisms in the pathogenesis of OSCC.