Comparative study of HPV prevalence in Japanese and North-east Chinese oral carcinoma

BACKGROUND: Human papillomavirus (HPV) plays a role in the development of oral carcinoma. However, the reported prevalence of HPV in oral carcinoma has varied widely. METHODS: The prevalence of HPV 16, 18 and 33 was investigated in Japanese and North-east Chinese oral squamous cell carcinomas (OSCCs) with polymerase chain reaction (PCR). The expression of p53 protein was examined immunohistochemically. RESULTS: HPV 16 and 18 were detected in 7 (23.3%) and 10 (33.3%) of 30 Japanese and 11 (36.7%) and 5 (16.7%) of 30 Chinese samples, respectively. HPV 16 and 18 coinfection was detected in 3/30 Japanese and 2/30 Chinese samples. HPV 33 was not detected. There was no significant correlation between HPV 16 and 18 and the sites, gender, age and histological grade. The prevalence of both HPV 16 and 18 was similar and higher in the Japanese and North-east Chinese samples (46.7% each). HPV 16 or/and 18 infection or/and p53 overexpression were in 22 (73.3%) of 30 Japanese samples and 24 (80.0%) of 30 North-east Chinese samples, respectively. CONCLUSIONS: HPV 16/18 infection or/and p53 overexpression may play an important role in developing some OSCCs. and the presence of HPV sequences and mutant p53 are not necessarily mutually exclusive.     

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.     

口腔鳞癌中HPV感染及其对p5 3改变影响的研究

目的：探讨高危型人乳头状瘤病毒（ＨＰＶ）在口腔鳞癌中的感染情况及其对Ｐ５３蛋白表达和ｐ５３突变的影响。方法：采用免疫组化和ＰＣＲ－ＳＳＣＰ方法，分别检测４０例来癌中高危型ＨＰＶＥ６蛋白表达、Ｐ５３蛋白表达和ｐ５３基因突变的情况。结果：９例ＨＰＶＥ６蛋白染色阳性，阳性率２２．５％（９／４０），与正常粘膜对照组有显著差异（Ｐ＝０．０２１）。ＨＰＶ阳性组中Ｐ５３蛋白表达率１１．１％（１／９），ＨＰＶ阴性     

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.     

p53 mutations and human papillomavirus infection in oral squamous cell carcinomas: correlation with overall survival.

PURPOSE: The study investigated the pattern of p53 gene mutations and human papillomavirus (HPV) infection concerning their relation to overall survival in patients with oral squamous cell carcinomas of the tongue and floor of the mouth. PATIENTS AND METHODS: The presence of HPV infection in 50 patients, and p53 gene mutations (42 patients from the same group) in the tumour specimens were analysed by polymerase chain reaction and single-stranded conformational polymorphism method. The follow-up period ranged from 12 to 48 (median 29) months. RESULTS: p53 mutations were identified in 11/42 tumours. HPV infection was detected in 32/50 cases, mostly HPV16 (10/32), HPV18 and HPV31 (6/32). A significantly higher incidence of HPV infection was found among smokers (p<0.05) and among patients with poor oral hygiene (p<0.01). The highest incidence of p53 mutations was detected in tumours of histological grade I and nuclear grade III. Patients with p53 mutation or with HPV infection had significantly shorter overall survival when compared with those that were without p53 mutations (p<0.01) or HPV infection (p<0.05). HPV-infected patients with p53 mutation had the worst prognosis when compared with patients with HPV infection only (p<0.01) or with patients negative for both HPV and p53 (p<0.01). CONCLUSION: The results stress once more the importance of HPV for the prognosis of survival of patients with squamous cell carcinoma of lower parts of the oral cavity. The presence of p53 mutations in HPV-infected tumours was associated with an even poorer prognosis for the patients.     

Detection of human papillomavirus DNA in benign oral squamous epithelial lesions in Venezuela

The polymerase chain reaction (PCR) was applied to the detection of human papillomavirus (HPV) infection in biopsies taken from clinically normal oral mucosa of 20 subjects and clinical lesions of 40 patients. PCR for HPV-DNA amplification was performed using consensus primers MYO9/MYO11 and subsequent typing for HPV of high and low oncogenic risk HPV types were identified by restriction enzyme analysis (restriction fragment length polymorphism, RFLP). The HPV viral genome was present in 55% (22/40) of the oral benign lesions (OBL) and in 10% (2/20) of the control samples. In the PCR+ OBL, we observed 90.9% of low oncogenic risk types (HPV-6 -13 and -32) and 9.1% of the samples had a mixed infection with low and high oncogenic types (HPV-6 and -16). In the control samples, we observed one patient with HPV-6 and another with HPV-6 and -16 in the same sample. All of the eight focal epithelial hyperplasia cases were positive for low risk HPV types (88% HPV-13 and 12.5% HPV-32). In conclusion, this study demonstrates a high incidence of HPV in oral benign lesions from Venezuelan patients.     

人乳头状瘤病毒16/18型、p53、p16蛋白在口腔疣状癌中的表达

目的　研究人乳头状瘤病毒 16 / 18型、p5 3、p16蛋白在口腔疣状癌中的表达状况 ,探讨它们在口腔疣状癌发生发展中的生物学意义。方法　采用SP免疫组化和原位杂交方法分别检测 8例正常口腔粘膜、13例疣状癌、10例高分化鳞癌、10例低分化鳞癌组织中HPV16 / 18E6、p5 3、p16蛋白和HPV16 / 18DNA的表达。结果　 1.疣状癌HPV16 / 18E6蛋白和p5 3蛋白阳性表达率均为 6 9.2 % (9/ 13) ,p16蛋白表达缺失率为 2 3.1% (3/ 13) ,过度表达率为 6 9.2 % (9/ 13) ,平均染色强度与高分化鳞癌和低化分鳞癌组比均有显著性差异 (P <0 .0 5 )。 2 .免疫组化方法检测HPV16 / 18E6蛋白与原位杂交方法检测HPV16 / 18DNA结果有良好的一致性。 3.疣状癌HPV16 / 18E6蛋白与p5 3蛋白、p5 3蛋白与p16蛋白表达之间无相关性 (P <0 .0 5 ) ,而HPV16 / 18E6蛋白与p16蛋白表达之间呈正相关 (P<0 .0 5 )。结论　 1.进一步证实HPV16 / 18型感染是口腔疣状癌的重要致病因子。 2 .疣状癌的发生过程中可能存在p5 3基因突变。 3.p16基因变异在疣状癌的发生中起一定作用 ,用疣状癌中p16蛋白过度表达与HPV16 / 18型感染有关。 4 .HPV16 / 18、p5 3、p16蛋白在疣状癌与高分化鳞癌、低分化鳞癌组织中的表达存在明显差异 ,证实疣状癌是一种独立类型     

Mutated and wild-type p53 expression and HPV integration in proliferative verrucous leukoplakia and oral squamous cell carcinoma

The frequencies of overexpression and mutation in the p53 tumor suppressor gene were examined in proliferative verrucous leukoplakia and oral squamous cell carcinoma with immunohistochemistry and single-strand conformation polymorphism analysis of DNA fragments amplified by polymerase chain reaction. Ten samples each of normal oral mucosa, proliferative verrucous leukoplakia, and squamous cell carcinoma were immunostained with antibodies against p53 protein; 8 of 10 cases of proliferative verrucous leukoplakia cases and 7 of 10 cases of oral squamous cell carcinoma were positive for p53 protein. Minimal staining was observed in normal oral tissues. The quantified labeling indexes demonstrated a range that corresponded to lesion progression. Single-strand conformation polymorphism analysis revealed p53 gene mutations within exons 5 to 8 in 40% (4 of 10) of the squamous cell carcinoma samples. Two of the 4 mutated squamous cell carcinoma samples lacked p53 expression. No p53 mutations were detected in proliferative verrucous leukoplakia tissues. Human papillomavirus 16 was identified in 2 of 7 p53 positive oral squamous cell carcinoma samples. Human papillomavirus 16 and 18 were identified in two of eight p53 positive proliferative verrucous leukoplakia samples. One p53 negative squamous cell carcinoma sample was positive for human papillomavirus 16 and had a mutation in exon 6 of the p53 gene. Human papillomavirus infection along with p53 expression plays a yet to be defined role in the pathogenesis of a limited number of cases of proliferative verrucous leukoplakia and squamous cell carcinoma. p53 Immunohistochemistry, p53 gene mutations, and human papillomavirus infection prevalence do not provide a means to differentiate between leukoplakia and carcinoma and do not provide a predictive test for progression of leukoplakia to carcinoma.     

The prevalence and distribution of human papillomavirus genotypes in oral epithelial hyperplasia: proposal of a concept

Background:  Evidence is accumulating for the aetiological role of human papillomavirus (HPV) in the pathogenesis of potentially malignant oral mucosal lesions and squamous cell carcinomas.
Methods:  Paraffin tissue sections from 49 patients with 'white patches' of the oral mucosa were investigated histologically, by broad-spectrum PCR followed by genotyping and chromogenic in situ hybridisation (CISH).
Results:  Histologically, 33 flat hyperplasias and 16 papillary hyperplasias were diagnosed. Twenty-two of 28 samples studied (78.6%) were positive for HPV DNA by PCR and six were negative. The following HPV types were detected in decreasing order of prevalence: HPV 35, HPV 6, HPV16, HPV 53, HPV 18, HPV 51 and HPV 55. Seventeen samples (60.7%) contained high-risk HPV DNA. Using CISH, ≥ 1 HPV signals were detected at least in a few epithelial cells in 95% of cases studied. All but one case were positive with the high-risk HPV probe and all HPV infections contained low viral load. Concordant positive results both by PCR and CISH were detected in 14 of 19 cases (73.7%) analysed.
Conclusions:  The high prevalence of HPV infection in hyperplastic 'white patches' of the oral mucosa supports the putative role of HPV at an early stage of oral carcinogenesis. These results further indicate that the majority of white oral mucosal lesions – flat, exophytic, wart-like or papillary proliferations – could be considered as the clinical manifestations of oral HPV infection. This finding has clinical relevance regarding therapy and patient management and may help in elucidating the role of HPV infection in oral carcinogenesis.     

Cancer genes alterations and HPV infection in oral squamous cell carcinoma

The aim of this study was to gain a better understanding of cancer genes contributing to oral squamous cell (OSCC) development and progression and correlate genetic changes to clinical parameters. Human papilloma virus (HPV) 16 detection is also included in the study. 60 samples of OSCC were analysed for c-erbB2 and c-myc amplification by dPCR, H-ras and p53 point mutations by PCR/SSCP. HPV was detected via amplification of its E1 and E6 genes. c-erbB2 was altered in 45%, c-myc in 35%, H-ras in 22% and p53 in 60% of samples. HPV was detected in 10% of cases. The frequency of p53 gene mutations showed a statistically significant association with tumour stage. Patients with c-erbB2 and H-ras alterations had lower survival than patients without these alterations. The number of detected genetic changes was remarkable but statistical association with tumour natural history was poor, indicating high clonal heterogeneity and multiple pathways of carcinogenesis.     

Low prevalence of HPV infection and its natural history in normal oral mucosa among volunteers on Miyako Island, Japan

OBJECTIVE: To investigate the prevalence of human papillomavirus (HPV) infection in normal oral mucosa, and to observe the natural history in the oral cavity in oral swab samples collected from healthy volunteers on Miyako Island, Okinawa, Japan. STUDY DESIGN: The prevalence of HPV infection in oral buccal mucosa cell scrapes collected between 2000 and 2002 from a cohort of 668 healthy volunteers was determined. HPV DNA was detected by consensus polymerase chain reaction (PCR) using MY09/MY11 primers followed by direct cycle sequencing. Just over 2 years later the HPV-positive participants were reevaluated. RESULTS: Of the 668 subjects, 662 samples were analyzed for HPV. HPV DNA was detected in 4 (0.6%) specimens. HPV type 16 (HPV16), HPV53, and HPV71, mucosal types, and HPV12, a cutaneous type, were all identified by direct sequencing. In the follow-up survey, the HPV71- and HPV12-positive participants again tested positive, while HPV DNA was not detected in the HPV16- and HPV53-positive participants. CONCLUSION: The results of this study among healthy individuals from Miyako Island suggest that oral HPV infection is uncommon. In this cohort, HPV71 and HPV12 were persistent, while HPV16 and HPV53 were transient in normal oral mucosa.     

p53 polymorphism, human papillomavirus infection in the oral cavity, and oral cancer

OBJECTIVES: Human papillomavirus (HPV) infection has emerged as a risk factor in oral carcinogenesis. An arginine-coding polymorphism of the tumor suppressor protein p53 at codon 72 is more readily degraded by the HPV oncoprotein E6. Our objective was to evaluate the association between p53 polymorphism at codon 72 and HPV infection in the oral cavity, as well as its association with oral cancer.Study Design: Oral squamous cells from 202 patients with oral cancer and 333 age-sex frequency matched controls were evaluated by polymerase chain reaction for the presence and type of HPV and for alleles of codon 72 in p53. Fisher exact test and chi(2) tests were used to evaluate the data. RESULTS: The p53 codon 72 polymorphism is not associated with HPV infection, whether comparing HPV-negative controls with HPV-positive controls or comparing HPV-negative cases with HPV-positive cases. Additionally, we found no association with the codon 72 polymorphism and oral cancer, whether comparing HPV-negative controls with HPV-negative cases or comparing HPV-positive controls with HPV-positive cases. CONCLUSIONS: There is no association between p53 codon 72 polymorphism and HPV infection or between the p53 polymorphism and the risk of oral cancer.     

口腔鳞癌HPV16、18感染与多基因表达的相关性研究

目的:检测口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)p53、Ki-67、Rb、p16和cyclin D1蛋白表达与高危型人类乳头状瘤病毒(human papilloma virus,HPV)感染的相互关系.方法:对73例OSCC组织标本,应用免疫组织化学ABC法检测病变组织中p...     

p53 gene mutations and HPV infection in primary head and neck squamous cell carcinomas do not correlate with overall survival: a long term follow-up study

We analyzed specimens of head and neck squamous cell carcinomas (HNSCC) from 110 patients for p53 gene mutations, and 92 of them for human papillomavirus (HPV) infection, in order to evaluate the prognostic significance of these factors by comparison with clinical follow-up data. Mutations within the exons 5 to 8 of the p53 gene were found in 48 tumors (44%). Sequencing revealed in most cases mis-sense mutations (16/21). Frequency of p53 gene mutations was not related to the tumor stage or the presence of lymph node metastases. Of the 46 tumors that were analyzed by immunohistochemistry. 26 stained positively (56%). The number of positively stained nuclei increased slightly with decreasing differentiation of the tumors, whereas no correlation was found between tumor stage and immunoreactivity. An infection with the high-risk HPV types 16 and 18 could be detected in 39/92 tumor specimens (42%.). Follow-up data were obtained from 99 patients within a range of 2 to 112 months. No dependence of overall survival on the presence of p53 gene mutations or HPV infection could be observed. The absence of statistically significant correlations between p53 gene mutation and progressive disease, however, does not deny its putative relevance in early phases of tumor development.     

Alterations of p16/CDKN2, p53 and ras genes in oral squamous cell carcinomas and premalignant lesions

Exons 1–3 of the p16/CDKN2 gene, exons 4–9 of the p53 gene and exons 1 and 2 of H-, K- and N- ras genes were screened for mutations by a combination of immunohistochemistry and single-strand conformational polymorphism (SSCP) analyses of polymerase chain reaction products from human surgical samples of both frank oral squamous cell carcinomas and premalignant lesions. The samples included 20 squamous cell carcinomas. 10 epithelial dysplasias and 10 epithelial hyperplasias. No identifiable gene mutations were detected in any of the dysplasias or hyperplasias, while 2 (10%) deletions and 2 (10%) mutations of p16/CDKN2, along with 5 (25%) p53 mutations were found in the advanced carcinomas, yielding characteristic p16/CDKN2 and p53 changes. A mutation in the K- ras gene was found in single carcinoma and dysplastic samples. From the data, it can be argued that p16/CDKN2 and p53 mutations are relatively late occurrences in human oral tumorigenesis and that genetic alterations of the ras genes may not play a significant role in squamous neoplasia.     

Human papilloma virus in erosive oral lichen planus

Several types of human papilloma viruses (HPV) have been associated with benign and malignant squamous cell tumours of mucosal epithelium. To identify HPV in erosive oral lichen planus (OLPe), considered as a premalignant lesion, tissues from 20 patients were examined by Southern blot hybridization with 32P-labeled HPV DNA probes. Type 11 was found in 6 of the lesions while HPV types 6, 16 and 18 were not detected in any of the tissues examined. Using a type-specific polymerase chain reaction (PCR) assay for HPV-6, 11, 16 and 18, HPV-11 was detected in 8 of the samples (all of those positive by Southern blot), and, in addition, HPV-6 was found in 5 samples and HPV-16 in 3 samples. Overall, by the more sensitive PCR assay, 65% of samples were positive for HPV DNA. The finding of HPV DNA in many of the samples using two different techniques indicates a high prevalence of HPV in the OLPe afflicted oral mucosa. However, the role of HPV in the pathogenesis of OLPe has yet to be determined.     

口腔鳞状细胞乳头状瘤组织中HPV DNA的原位杂交研究

目的 探讨人乳头瘤病毒（ｈｕｍａｎ ｐａｐｉｌｌｏｍａｖｉｒｕｓ，ＨＰＶ）感染与口腔鳞状细胞乳头状瘤（ｓｑｕａｍｏｕｓ ｃｅｌｌ ｐａｐｉｌｌｏｍａ，ＳＣＰ）的发生之间的关系。方法 应用地高辛标记的ＨＰＶ６／１１和ＨＰＶ１６／１８核酸探针分别在３０例口腔ＳＣＰ组织上进行原位杂交，检测口腔ＳＣＰ组织中ＨＰＶ ＤＮＡ的特征。结果 ＨＰＶ６／１１ ＤＮＡ阳性１６例（５３％），ＨＰＶ１６／１８ＤＮＡ未检出，ＨＰＶ６／１１ＤＮＡ阳性细胞多数分布在鳞状上皮的表层、中层和基底层。结论 原位杂交方法可以检测口腔ＳＣＰ组织中ＨＰＶ ＤＮＡ的存在并能准确组织定位，进一步支持ＨＰＶ６／１感染与口腔ＳＣＰ的发生密切相关。     

Comparison of HPV infection, p53 mutation and allelic losses in post-transplant and non-posttransplant oral squamous cell carcinomas

BACKGROUND: Oral squamous cell carcinoma (SCC) is increasingly found in transplant recipients, although little is known of the natural history of the disease or the mechanism underlying this increase. METHODS: In this article we describe the history of development of 5 oral post-transplant SCCs (PSCCs) and compare their genetic profiles to 34 non-posttransplant SCCs (NPSCCs). RESULTS: Of the five patients with PSCCs, 3 had bone marrow transplants and two, kidney. All three PSCCs from bone marrow recipients were preceded locally by graft-vs.-host disease (GVHD). Two of the GVHD were biopsied and demonstrated dysplasia. Similar frequencies of loss of heterozygosity (LOH) occurred in PSCCs and NPSCCs at 3p, 9p, 17p and 8p, with lower frequencies in PSCCs at 4q (39% vs. 0%), 11q (53% vs. 20%) and 13q (45% vs. 20%), although the latter were not significantly different. Only 1 PSCC had a p53 mutation, compared to historical values of 40-60% for NPSCC. Interestingly, human papillomavirus (HPV) DNA was detected in 3 (60%) PSCCs, in comparison to only 4 (12%) of the 34 NPSCCs (P = 0.0346). CONCLUSIONS: Dysplasia in oral GVHD may be a strong indicator of cancer risk and should not be regarded as reactive changes to lichenoid mucosites. The low level of p53 mutation and increased HPV infection support the involvement of HPV in the development of PSCC, while the similarity in LOH patterns suggests that other aspects of carcinogenesis may be comparable in these two types of SCCs.     

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.