The Aetiological Role of Human Papillomavirus in Oesophageal Squamous Cell Carcinoma: A Meta-Analysis

Background

The aetiological role of human papillomavirus (HPV) in oesophageal squamous cell carcinoma (OSCC) has been widely researched for more than three decades, with conflicting findings. In the absence of a large, adequately powered single case-control study, a meta-analysis of all available case-control studies is the most rigorous way of identifying any potential association between HPV and OSCC. We present the first global meta-analysis of case-control studies investigating the role of HPV in OSCC.

Methods

Case-control studies investigating OSCC tissue for presence of HPV DNA were identified. 21 case-control studies analyzing a total of 1223 cases and 1415 controls, met our inclusion criteria. HPV detection rates were tabulated for each study and all studies were assessed for quality. The random effects method was used to pool the odds ratios (OR).

Results

From all OSCC specimens included in this meta-analysis, 35% (426/1223) were positive for HPV DNA. The pooled OR for an HPV-OSCC association was 3.04 (95% CI 2.20 to 4.20). Meta-regression analysis did not find a significant association between OR and any of the quality domains. Influence analysis was non-significant for the effect of individual studies on the pooled estimate. Studies conducted in countries with low to medium OSCC incidence showed a stronger relationship (OR 4.65, 95% CI 2.47 to 8.76) than regions of high OSCC incidence (OR 2.65, 95% CI 1.80 to 3.91).

Conclusions

Uncertainty around the aetiological role of HPV in OSCC is due largely to the small number and scale of appropriately designed studies. Our meta-analysis of these studies suggests that HPV increases the risk of OSCC three-fold. This study provides the strongest evidence to date of an HPV-OSCC association. The importance of these findings is that prophylactic vaccination could be of public health benefit in prevention of OSCC in countries with high OSCC incidence.     

Human Papillomavirus Infection in 674 Chinese Patients with Laryngeal Squamous Cell Carcinoma

Objectives

Previous reports suggest a strong association between human papillomavirus (HPV) and the etiology of laryngeal squamous cell carcinoma (LSCC). However, clinical data regarding the HPV infection rate among LSCC patients remain largely inconsistent.

Methods

In total, 674 LSCC patients from three major hospitals in Shanghai were enrolled in this study. We determined the patients'' HPV infection status using immunohistochemistry and the GenoArray HPV genotyping assay and calculated their long-term survival rate using the Kaplan-Meier method.

Results

The total P16-positive rate according to immunostaining results was 7.57% (51/674). None of the P16-negative patients were HPV-positive according to the HPV genotyping test. The rate of HPV infection among patients with LSCC was 4.9% (33/674). HPV infection was more common among nonsmokers (P<0.05), nondrinkers (P<0.05), and patients with supraglottic LSCC (P<0.05). Of the 33 HPV-positive patients, 28 (84.8%) were infected with HPV-16, 2 with HPV-18, 1 with HPV-31, 1 with HPV-33 and 1 with HPV-45. The 3-year overall survival rate and progression-free survival rate were higher in HPV-positive than HPV-negative patients, but the difference was not statistically significant (76.3% vs. 70.7%, P = 0.30 and 65.1% vs. 58.3%, P = 0.37, respectively).

Conclusion

HPV was not a main causal factor in LSCC carcinogenesis in this Chinese population. HPV infection did not alter patients'' overall survival or progression-free survival rates in this study.     

Interaction between Alcohol Consumption and CYP 2C19 Gene Polymorphism in Relation to Oesophageal Squamous Cell Carcinoma

Objectives

The purpose of this study is to explore the relationship between the interactions of CYP2C19 gene polymorphisms and several environmental factors and oesophageal squamous cell carcinoma (OSCC).

Methods

In a case-control study of OSCC patients (n = 350) and healthy controls (n = 350), we investigated the roles of polymorphism in the CYP2C19 gene by the use of polymerase chain reaction - restriction fragment length polymorphism (PCR – RFLP) analysis.

Results

The CYP2C19^*3 AG+AA genotype was significantly more prevalent in OSCC patients (10.0% versus 3.43%; P<0.01). Multiple logistic regression analysis showed drinking (OR: 5.603, 95% CI: 3.431–11.112; P = 0.005) and smoking (OR: 4.341, 95% CI: 3.425–10.241; P = 0.001) was the independent risk factor of OSCC respectively, and there were significant interaction between CYP2C19^*3 and drinking (OR: 8.747, 95% CI: 6.321–18.122; P = 0.009).

Conclusions

The CYP2C19^*3 polymorphism and OSCC were synergistically and significantly associated in Chinese Han patients.     

Human Papillomavirus (HPV) Prevalence in Nasal and Antrochoanal Polyps and Association with Clinical Data

Objectives

The pathogenesis of sinonasal polyposis remains unclear, in spite of several investigative approaches. Antrochoanal polyps, a subgroup of sinonasal polyposis along with allergic- and chronic-inflammatory nasal polyps, mostly originate from the maxillary sinus and develop as a unilateral, pedunculated mass towards the nasopharynx. The human papillomavirus (HPV) is discussed as a possible causative and influencing factor in development and progression of sinonasal polyposis. This study aims to elucidate HPV frequency in nasal polyps and antrochoanal polyps.

Materials and Methods

Genomic DNA from 257 tissue specimens (166 nasal polyps, 39 antrochoanal polyps and 52 nasal turbinates) was subjected to three different established HPV- polymerase chain reaction assays, testing for 37 low- and high-risk HPV. In addition, immunohistochemical analyses for HPV16 were carried out, as well as immunohistochemistry and western blots of p16, a biomarker for HPV induced cancer.

Results

HPV-DNA was detected in 53.8% of antrochoanal polyps, 15.1% of nasal polyps, and 5.8% of nasal turbinates. HPV16 was the predominant type with a detection rate of 76% in nasal polyps and 62% in antrochoanal polyps. Immunohistochemically, HPV positive tissues stained positive for HPV16 antigens and p16 in epithelial cell layers. No significant p16 overexpression was traceable in antrochoanal polyps, nasal polyps and nasal turbinates by western blot. There was no correlation of HPV-status with sex, age, smoking, alcohol consumption or allergic background.

Conclusion

The present study shows a significant frequency of high-risk type HPV16 in antrochoanal polyps. Absence of oncogenic transformation or correlation of the HPV-status with clinical data suggests a latent superinfection, possibly because of anatomical proximity to the oropharynx.     

HLA Class I and II Expression in Oropharyngeal Squamous Cell Carcinoma in Relation to Tumor HPV Status and Clinical Outcome

HPV-DNA positive (HPV_DNA+) oropharyngeal squamous cell carcinoma (OSCC) has better clinical outcome than HPV-DNA negative (HPV_DNA-) OSCC. Current treatment may be unnecessarily extensive for most HPV+ OSCC, but before de-escalation, additional markers are needed together with HPV status to better predict treatment response. Here the influence of HLA class I/HLA class II expression was explored. Pre-treatment biopsies, from 439/484 OSCC patients diagnosed 2000-2009 and treated curatively, were analyzed for HLA I and II expression, p16^INK4a and HPV DNA. Absent/weak as compared to high HLA class I intensity correlated to a very favorable disease-free survival (DFS), disease-specific survival (DSS) and overall survival (OS) in HPV_DNA+ OSCC, both in univariate and multivariate analysis, while HLA class II had no impact. Notably, HPV_DNA+ OSCC with absent/weak HLA class I responded equally well when treated with induction-chemo-radiotherapy (CRT) or radiotherapy (RT) alone. In patients with HPV_DNA- OSCC, high HLA class I/class II expression correlated in general to a better clinical outcome. p16^INK4a overexpression correlated to a better clinical outcome in HPV_DNA+ OSCC. Absence of HLA class I intensity in HPV_DNA+ OSCC suggests a very high survival independent of treatment and could possibly be used clinically to select patients for randomized trials de-escalating therapy.     

新疆维族妇女感染的HPV 型别分布的研究

目的:探讨人乳头瘤状病毒(Human papillomavirus,HPV)在新疆南部维族妇女人群中的型别分布情况。方法:以年龄在30-59岁的新疆伽师县夏普吐勒乡维吾尔族妇女人群为基础进行筛查,签署知情同意书后,采集受试者宫颈脱落细胞,利用PCR和基因芯片技术检测HPV DNA并分型。结果:共2473名妇女入选。HPV总的感染率为9.1%,高危型中HPV-16的感染率最高为6.9%,其他高危型的感染率从高到低依次为:HPV-59、HPV-56、HPV-18、HPV-33、HPV-58、HPV-51、HPV-31、HPV-45、HPV-52、HPV-68、HPV-35、HPV-39。低危型中HPV11感染率最高,其他低危型的感染率从高到低依次为HPV-42、HPV-43、HPV-6、HPV-53、HPV-66、HPV-73。HPV-44、-83、-MM4没检测到。多重感染率为34.2%。结论:新疆维吾尔族妇女人群中以HPV16感染为主,其次为HPV59、56、18、33等。HPV59可能是新疆维吾尔族妇女较易感染的类型。体现了新疆维吾尔族妇女感染HPV的特殊性。     

Reduced Prevalence of Oral Human Papillomavirus (HPV) 4 Years after Bivalent HPV Vaccination in a Randomized Clinical Trial in Costa Rica

Background

Human papillomavirus (HPV) infection, particularly with type 16, causes a growing fraction of oropharyngeal cancers, whose incidence is increasing, mainly in developed countries. In a double-blind controlled trial conducted to investigate vaccine efficacy (VE) of the bivalent HPV 16/18 vaccine against cervical infections and lesions, we estimated VE against prevalent oral HPV infections 4 years after vaccination.

Methods and Findings

A total of 7,466 women 18–25 years old were randomized (1∶1) to receive the HPV16/18 vaccine or hepatitis A vaccine as control. At the final blinded 4-year study visit, 5,840 participants provided oral specimens (91·9% of eligible women) to evaluate VE against oral infections. Our primary analysis evaluated prevalent oral HPV infection among all vaccinated women with oral and cervical HPV results. Corresponding VE against prevalent cervical HPV16/18 infection was calculated for comparison. Oral prevalence of identifiable mucosal HPV was relatively low (1·7%). Approximately four years after vaccination, there were 15 prevalent HPV16/18 infections in the control group and one in the vaccine group, for an estimated VE of 93·3% (95% CI = 63% to 100%). Corresponding efficacy against prevalent cervical HPV16/18 infection for the same cohort at the same visit was 72·0% (95% CI = 63% to 79%) (p versus oral VE = 0·04). There was no statistically significant protection against other oral HPV infections, though power was limited for these analyses.

Conclusions

HPV prevalence four years after vaccination with the ASO4-adjuvanted HPV16/18 vaccine was much lower among women in the vaccine arm compared to the control arm, suggesting that the vaccine affords strong protection against oral HPV16/18 infection, with potentially important implications for prevention of increasingly common HPV-associated oropharyngeal cancer.ClinicalTrials.gov, Registry number {"type":"clinical-trial","attrs":{"text":"NCT00128661","term_id":"NCT00128661"}}NCT00128661     

人乳头瘤病毒在宫颈鳞癌宫旁组织中的表达及意义

为研究高危型人乳头瘤病毒(high-risk human papillomavirus,hr-HPV)在宫颈鳞癌宫旁组织中的表达及意义,采用免疫组织化学技术对51例宫颈鳞癌患者术后宫旁组织不同位点的hr-HPV表达情况进行检测,应用重复测量设计的方差分析及单因素分析方法进行统计。hr-HPV在宫旁组织中的表达随着距宫颈距离的增加呈明显的梯度下降趋势(P0.05),在不同临床分期、宫颈浸润深度、淋巴结转移组内表达存在差异(P0.05),当临床分期ⅠB1期、宫颈浸润深度≤1/2以及无淋巴结转移时,hr-HPV在距离宫颈2 cm处主韧带、骶韧带及距离宫颈3 cm处阴道中表达开始与正常对照组无差异(P0.05)。结果表明,部分早期宫颈鳞癌患者可选择Ⅱ型改良式根治性子宫切除术,但阴道切除仍推荐3 cm。     

Prevalence and risk factors of Human Papillomavirus (HPV) infection in southern Chinese women - a population-based study

Background

Persistent high-risk type Human papillomavirus (HPV) infection is recognized as a necessary cause of cervical cancer. This study aimed to compare the HPV prevalence and risk factors between women residing in Hong Kong (HK) and Guangzhou (GZ) region of China.

Methodology/Principal Findings

A total of 1,570 and 1,369 women were recruited from HK and GZ, respectively. The cytology samples were collected and tested for HPV infection. The overall and type-specific HPV prevalence and the potential risk factors for acquisition of HPV infection were studied. Women with normal cytology in the GZ cohort had significantly higher HPV prevalence (10%) than those in the HK cohort (6.2%, p<0.001). The patterns of the age-specific HPV prevalence were also different between the two cohorts. In the HK cohort, women at the age of 20–29 years old had the highest prevalence and a second peak was observed in the age of ≥60 years old. In the GZ cohort, the highest HPV prevalence was also observed in 20–29 years old but declined as the age increased and a second peak was not seen. HPV16 and HPV52 were the most common high-risk types found in the HK and GZ cohorts, respectively. Age was the most consistently observed independent risk factor for HPV infection in the HK, while the number of sexual partners had association in the GZ cohort.

Conclusions/Significance

Our study provides the current status and the epidemiological characteristics of HPV prevalence in Southern Chinese women. The results strongly suggested that population education and the effective cervical cancer screening would be vital in the prevention of cervical cancer.     

人尿道(阴茎)鳞癌上皮细胞(HUS-98)cDNA文库的构建及鉴定

为了进一步分离人尿道(阴茎)鳞癌组织特异性表达基因和鳞癌特异性相关基因,采用SMART技术,构建了人尿道 (阴茎)鳞癌上皮细胞cDNA文库,从人尿道(阴茎)鳞癌上皮细胞中分离总RNA并纯化mRNA,利用经修饰的oligo(dT)引物 合成cDNA第一链,利用SMART核苷酸作为cDNA第一链在mRNA5′端延伸出去的模板,采用LD-PCR合成双链cDNA,双链 cDNA经酶切和过柱分级分离后,克隆入λTriplEx2载体后经体外包装而成cDNA文库。结果表明原始人尿道(阴茎)鳞癌上 皮cDNA文库获得1.57×107个重组子,重组率达到98%。文库扩增后,滴度达到4.0×109pfu/ml,插入cDNA平均长度为2.5kb。 构建的人尿道(阴茎)鳞癌上皮cDNA文库具有良好的质量,该cDNA文库为进一步筛选鳞癌抑癌基因及鳞癌特异性表达基因 奠定了基础。     

抗人尿道高分化鳞癌单克隆抗体的研制

用本室建立的人尿道高分化鳞状上皮癌细胞株(Hus-98)免疫Balb/C小鼠,通过细胞杂交技术及ELISA方法进行筛选,SP免疫组织化学方法进行鉴定,获得10株能稳定分泌、选择性强、效价高的杂交瘤细胞株;同时从乳腺癌组织中提出肿瘤相关抗原行免疫印记实验(Western-blot)。结果6株经4次克隆能稳定分泌抗体Ig,免疫组化显示这些抗体是针对来源于上皮的鳞癌、腺癌细胞膜和细胞浆的,而癌旁正常组织及胎儿组织呈阴性反应;通过western-blot结果显示肿瘤相关抗原分子量为46.414kb。     

Therapeutic strategies of different HPV status in Head and Neck Squamous Cell Carcinoma

Head and neck squamous cell carcinoma (HNSCC) is the 9^th most common malignant tumor in the world. Based on the etiology, HNSCC has two main subtypes: human papillomavirus (HPV) -related and HPV-unrelated. HPV-positive HNSCC is more sensitive to treatment with favorable survival. Due to the different biological behaviors, individual therapy is necessary and urgently required to deduce the therapeutic intensity of HPV-positive disease and look for a more effective and toxicity-acceptable regimen for HPV-negative disease. EGFR amplification and PI3K/AKT/mTOR pathway aberrant activation are quite common in HPV-positive HNSCC. Besides, HPV infection alters immune cell infiltrating in HNSCC and encompasses a diverse and heterogeneous landscape with more immune infiltration. On the other hand, the chance of HPV-negative cancers harboring mutation on the P53 gene is significantly higher than that of HPV-positive disease. This review focuses on the updated preclinical and clinical data of HPV-positive and HPV-negative HNSCC and discusses the therapeutic strategies of different HPV status in HNSCC.     

人乳头瘤病毒(HPV)基因芯片的研究

探讨将基因芯片与限制性显示技术相结合对HPV进行基因检测和分型的方法。分离HPV6,11,16和18型的基因片段作为探针,纯化后应用PixSys 5500点样仪将其打印在氨基包被的玻片上制作HPV基因芯片,对HPV样品进行荧光标记后与芯片杂交,经清洗和干燥后对芯片进行扫描和结果分析。对HPV基因检测芯片的制作与检测的实验条件进行了初步研究,并对应用HPV基因芯片进行分型做了初步探讨。建立的检测芯片实验方法可行,并且显示了在HPV分型中的应用前景。     

Human Papillomavirus (HPV) 16 E6 seropositivity is elevated in subjects with oral HPV16 infection

IntroductionHuman Papillomavirus (HPV) 16 E6 serum antibodies are common in people with HPV-related oropharyngeal cancers (HPV-OPC), but not the general population. We explored HPV16 seroprevalence in people with and without oral HPV16 infection, the cause of HPV-OPC.MethodsOral rinse samples were collected semiannually and tested for 36 types of HPV DNA by PCR. HPV16 E6 serum antibodies were tested at the visit of first oral HPV detection in participants with prevalent (n = 54), or incident (n = 39) oral HPV16 DNA; or at baseline in matched participants with no oral HPV16 DNA (n = 155) using multiplex serology assay. Predictors of seropositivity were examined using logistic regression.ResultsHPV16 E6 seropositivity (7.5% vs 0.7%; p = 0.005) but not seropositivity to the other HPV16 antigens, was significantly more common in those with than without oral HPV16 infection. There were only 8 HPV16 E6 seropositive participants, but oral HPV16 DNA remained a strong predictor of E6 seropositivity after adjustment for other risk factors (aOR = 14.6 95%CI, 1.7–122.5). Seroprevalence was similar in those with prevalent (7.4%; 4/54), and incident (7.7%; 3/39) oral HPV16 infection (p = 1.00). E6 seroprevalence was associated with reduced oral HPV16 clearance, but was not statistically significant (HR = 0.65 95% CI, 0.16–2.70).Seropositive participants were primarily male (87.5%), HIV-positive (75.0%; median CD4 cell-count of 840) and had oral HPV16 DNA (87.5%). History of an HPV-related cancer (0/8) or HPV-related anogenital dysplasia (1/8) was rare, and 4 participants had recent screening showing no anogenital dysplasia.DiscussionHPV16 E6 seropositivity was higher among people with than without oral HPV16 infection, despite no known anogenital disease in these participants.     

Human Papillomavirus in Head and Neck Squamous Cell Carcinoma of Unknown Primary Is a Common Event and a Strong Predictor of Survival

Background

The purpose of this study was to examine the prevalence of human papillomavirus (HPV) in patients with head and neck squamous cell carcinoma of unknown primary (CUP).

Methods

All patients diagnosed with and treated for CUP between January 1, 2000, and June 1, 2011, at two Danish medical centers were included. All patients received a thorough diagnostic work-up, including FDG-PET, before being diagnosed as CUP. We determined the HPV status in all patients using a combination of HPV DNA PCR and p16 stain. In addition, clinical information on the study patients was retrieved from clinical records.

Results

Of the identified 60 patients with CUP, 13 were shown to be positive for HPV DNA, amounting to 22% of the study population. In addition, we were able to show a clear disease-free and overall-survival benefit in the HPV-positive group, with a hazard ratio of 0.16 (95% CI: 0.038–0.67) for over-all survival. This survival benefit was also apparent when adjusted for advanced age in a multivariate Cox regression analysis.

Conclusion

A fairly large percentage of CUP cases are HPV-related, and because this is related to both the location and prognosis, we recommend HPV testing as part of the diagnostic work-up.     

Oral Squamous Cell Carcinoma in a Pigtailed Macaque (Macaca nemestrina)

An adult, gravid, female pigtailed macaque (Macaca nemestrina) presented for facial swelling centered on the left mandible that was approximately 5 cm wide. Differential diagnoses included infectious, inflammatory, and neoplastic origins. Definitive antemortem diagnosis was not possible, and the macaque''s condition worsened despite supportive care. Necropsy findings included a mandibular mass that was locally invasive and expansile, encompassing approximately 80% of the left mandibular bone. The mass replaced portions of the soft palate, hard palate, sinuses, ear canal, and the caudal–rostral calvarium and masseter muscle. Histologically, the mass was a neoplasm that was poorly circumscribed, unencapsulated, and infiltrative invading regional bone and soft tissue. The mass consisted of polygonal squamous epithelial cells with intercellular bridging that breached the epithelial basement membrane and formed invasive nests, cords, and trabeculae. The mitotic rate averaged 3 per 400× field of view, with occasional bizarre mitotic figures. Epithelial cells often exhibited dyskeratosis, and the nests often contained compact lamellated keratin (keratin pearls). The neoplasm was positive via immunohistochemistry for pancytokeratin, variably positive for S100, and negative for vimentin, smooth muscle actin, and desmin. The gross, histologic, and immunohistochemical findings were consistent with an aggressive oral squamous cell carcinoma. The neoplasm was negative via PCR for papilloma virus. In general, neoplasia in macaques is rare. Although squamous cell carcinomas are one of the most common oral neoplasia in many species, to our knowledge this case represents the first reported oral squamous cell carcinoma in a pigtailed macaque.Abbreviation: SCC, squamous cell carcinomaSquamous cell carcinomas (SCC) are one of the most commonly reported oral tumors. They are characterized as firm, nodular to irregular, soft-tissue masses that are often ulcerated.⁶ These tumors are frequently highly invasive to local bone and muscle and occasionally metastasize to local and regional lymph nodes.⁶ Histologically, SCC are characterized by keratin pearls, intercellular bridges, and positive cytokeratin staining on immunohistochemistry.^6,18 SCC have been associated with carcinogen exposure (such as bracken fern toxicosis in cattle), actinic radiation, and rarely with papillomatosis.⁸In general, neoplastic diseases are rare in nonhuman primates, and SCC and lymphoma are the 2 most commonly reported oral neoplasms in these species.³ SCC have most commonly been reported in rhesus macaques (Macaca mullata) and baboons (Papio spp.) among nonhuman primate species.⁹ In rhesus macaques, SCC has occurred in the oral cavity,⁹ integument,^9,22 esophagus,⁹ stomach,²¹ lung,^9,13 prepuce–penis,¹⁰ cervix,⁹ uterus,⁹ and eye.⁹ These neoplasms have also been reported to occur in cynomolgus macaques,^14,15,17,19 marmosets, squirrel monkeys, tree shrews, capuchins, tamarins, black spider monkeys, sooty mangabies, a spectacled langur, and an orangutan.⁹ No report describing SCC in a pig-tailed macaque has been published previously. The oral cavity is the most common site of SCC in nonhuman primates, and metastasis occurs in approximately 23% of cases.⁹ The average age at diagnosis of oral SCC in rhesus macaques is 17.6 y.²² In baboons, SCC is the third most common neoplasm, after intestinal adenocarcinoma and lymphosarcoma.⁴ The following case report describes an oral SCC in a pregnant pig-tailed macaque.     

Cell Surface Oligonucleotide-Binding Proteins of Human Squamous Carcinoma A431 Cells

Abstract

Affinity modified with Flu-DAP-p(N)₁₆degU oligonucleotide-binding proteins were isolated by affinity chromatography using Ultrogel A2 - anti fluorescein antibodies. After separation by SDS-PAGE the proteins with molecular masses about 68 kDa were MS/MS sequenced and identified as keratin K1, keratin K10, keratin K2e and albumin.     

An Animal Explant Model for the Study of Human Cutaneous Squamous Cell Carcinoma

We established a human tissue explant model to facilitate study of cutaneous squamous cell carcinoma. We accomplished this by implanting debulked SCC, from surgical discard, into nude rats. Human SCC remained viable and continued to proliferate for at least 4 weeks and showed evidence of neovascularization. At 4 weeks, SCC implants showed a trend toward increased PCNA positive cells compared to fresh SCC cells/mm² tissue) supporting continued proliferation throughout engraftment. Von Willebrand''s Factor (VWF) positive cells were found within implants and likely represented rat vessel neovascularization. Human Langerhans'' (Langerin+) cells, but no T cells (CD3+, CD8+, FoxP3+), macrophages (CD163), or NK cells (NKp46), were present in SCC implants at 4 weeks. These findings support the possibility that LCs fail to migrate from cutaneous SCC and thus contribute to lack of effective antitumor response. Our findings also provide a novel model system for further study of primary cutaneous SCC.