Marker For Human Papillomavirus Infection Research Articles

Analysis of the anatomical distribution of HPV genotypes in head and neck squamous papillomas.

Squamous papillomas (SPs) of the head and neck are usually benign lesions associated with human papilloma virus (HPV) infection. However, the reported HPV detection rates vary widely, especially with respect to anatomical distribution. The etiology of SPs in the head and neck remains unclear; analyzing HPV genotypes of SPs based on anatomical sites could assist in clarifying the pathogenesis of SPs in the head and neck. Therefore, the aim of this study was to review the prevalence, subtypes, and anatomical distribution of HPV in head and neck SPs at a hospital in China; we also investigated whether p16, a marker of HPV infection in oropharyngeal carcinoma, could serve as a surrogate marker for HPV in head and neck SPs. The presence of HPV DNA of 23 types (5 low-risk HPV and 18 high-risk HPV types) was detected via real-time PCR. p16 immunohistochemistry was performed using SP sections. Age, sex, anatomical location, and HPV subtype were recorded for each case. In total, 105 SPs were identified, including 47 in the larynx, 42 in the pharynx, 6 in the external auditory canal (EAC), 5 in the oral cavity, and 5 in the nasal cavity. HPV was found in 57 (54.3%) cases, with the highest positivity rate in the larynx (46/47; 97.9%). Only 5/42 (11.9%) patients showed HPV positivity in the pharynx. HPV incidence was highly dependent on the anatomical site. SPs in the larynx and EAC were more likely to carry HPV than those in other anatomical sites. High-risk HPV infections were rarely associated with SPs in the head and neck region. The sensitivity and specificity of p16 immunohistochemistry for HPV infection were 88% and 96%, respectively. There may be an association between p16 and HPV infection in head and neck SPs, but further studies are needed to validate this assertion.

Epigallocatechin gallate, folic acid, vitamin B12, and hyaluronic acid significantly increase apoptosis and p53 expression in HeLa cells.

The human papilloma virus (HPV) is the etiological agent of cervical cancer in more than 95% of cases worldwide. Although most HPV infections clear up on their own and most pre-cancerous lesions spontaneously resolve, in some cases, they can persist, leading to lesions which may progress towards invasive cervical cancer. We evaluated the effects of the association of epigallocatechin gallate (EGCG) + folic acid (FA) + vitamin B12 (B12) + hyaluronic acid (HA) on HPV-positive cervical cancer cells (HeLa). The association of EGCG + FA + B12 + HA induced a significant increase of apoptosis and p53 gene expression with a concomitant decrease of E6/E7 gene expression, a marker of HPV infection. This study provides for the first-time evidence on the potential additive activity of EGCG + FA + B12 + HA in counteracting HPV infection, by increasing apoptosis and p53 expression in HPV-infected cervical HeLa cells.

The Proportion of p16 Expression in Penile Squamous Cell Carcinoma Based on Immunohistochemistry Examination in the Balinese Population

Background: : Penile cancer is a rare type of cancer in men and was the highest in Bali compared to other regions in Indonesia. Around 95% of malignancies in the penis are squamous cell carcinoma (SCC) type. One of the risk factors associated with penile squamous cell carcinoma is an infection caused by Human Papillomavirus (HPV). Human Papillomavirus E6 and E7 oncoproteins play a significant role in penile squamous cell carcinoma carcinogenesis. Overexpression of p16 protein can be used as a marker of HPV infection. This study aims to determine the expression of p16 protein in penile squamous cell carcinoma.Methods: This study was conducted to determine the prevalence of p16 protein expression was assessed by immunohistochemical examination from paraffin block tumors of penile cancer patients. Expression of p16 was observed to be positive when stained with a strong brown color in the nucleus and cytoplasm of tumor cells. The analysis method used in this study is a descriptive statistic to provide a general description of the characteristics of each research variable. The data were processed using SPSS to determine the frequency and median of the age of patients with penile squamous cell carcinoma and the prevalence of p16 protein expression in cell penile squamous cell carcinoma. The data shown in this study are the age range and prevalence of p16 protein expression in penile squamous cell carcinoma using the prevalence table.Results: This study used 142 penile squamous cell carcinoma samples derived from biopsy, partial, and total penectomy. The results of the IHC examination showed that the expression of p16 protein was positive in penile cancer tumor cells in 68 cases (47.9%), with the highest age distribution in the 50–59-year age group (28.2%).Conclusions: Further research is needed to confirm the correlation between HPV infection and p16 protein overexpression in penile squamous cell carcinoma, including HPV genotyping

Rectal Condyloma Acuminatum

A woman between 70 and 80 years of age with a history of cervical cancer presented with changes in bowel habit. The patient took prednisolone for cardiac sarcoidosis. Colonoscopy revealed a 20-mm flat, whitish lesion with diffuse reddish speckles in the lower rectum (Figure 1). Based on the medical history and endoscopic findings, we suspected rectal condyloma acuminatum. However, biopsy results could not provide a definitive diagnosis; thus, we performed endoscopic submucosal dissection for excisional biopsy. The resected specimen revealed squamous epithelial cells with vacuolated cytoplasm (Figure 2). P16 staining, a marker of human papillomavirus infection, was diffusely stained. A final diagnosis of condyloma acuminatum was made. The patient was discharged uneventfully. No recurrence was observed 8 months after treatment.Figure 2Resected specimen showing squamous cells with vacuolated cytoplasm.View Large Image Figure ViewerDownload Hi-res image Download (PPT) Condyloma acuminatum, a relatively common disease especially in young adults, has an annual incidence ranging from 160 to 289 per 100,000 individuals.1Patel H. Wagner M. Singhal P. Kothari S. Systematic review of the incidence and prevalence of genital warts.BMC Infect Dis. 2013; 13: 39Crossref PubMed Scopus (230) Google Scholar Condyloma acuminatum is a viral wart caused by human papillomavirus infection and can also be observed in immunocompromised patients, such as those taking immunosuppressive agents. Generally, condyloma acuminatum is detected in the perineum, labia, vagina, penis, perianal area, and anal canal. However, it rarely crosses the dentate line to grow into the rectum. Condyloma acuminatum has malignant potential, often developing into atypical epithelium or carcinoma; thus, surgical resection is the first-line treatment. However, recent reports have shown the efficacy of endoscopic submucosal dissection as a curative, minimally invasive option.2Matsumoto K. Sakamoto N. Murakami T. et al.Long-term relapse-free treatment with endoscopic submucosal dissection combined with magnifying narrow-band imaging for a pregnant patient with flat-type condyloma acuminatum: a case report.Clin J Gastroenterol. 2021; 14: 439-445Crossref Scopus (2) Google Scholar There is no consensus on surveillance of rectal condyloma acuminatum; thus, it is reasonable to apply the surveillance recommendations for anal condyloma acuminatum, with follow-ups every 6 to 12 months.3Long K.C. Menon R. Bastawrous A. Billingham R. Screening, surveillance, and treatment of anal intraepithelial neoplasia.Clin Colon Rect Surg. 2016; 29: 57-64Crossref Scopus (30) Google Scholar The authors report no potential competing interests.

Rising incidence of HPV positive oropharyngeal cancer in Taiwan between 1999 and 2014 where betel nut chewing is common

BackgroundThe incidence of human papillomavirus (HPV) positive oropharyngeal cancer (OPC) is rising but HPV negative OPC is decreasing in Western countries. In Taiwan, the incidence of HPV negative OPC is common but the incidence of HPV positive OPC remains unknown. The objective of this study is to estimate the incidence trend and the survival of HPV positive OPC in Taiwan.MethodsBetween 1999 and 2014, primary tumor tissues from 425 incident OPCs were obtained from 5 medical centers in Taiwan. 408 OPCs were evaluated by the EasyChip HPV genotyping (King-Car, I-Lan, Taiwan) and 369 OPCs by p16 staining. The clinical data were retrospectively obtained from the medical records.ResultsIn our study, 29% of OPCs were HPV positive. The percentage of HPV positive OPC was stable from 1999 to 2014 (25% (1999–2002), 30% (2003–2006), 30% (2007–2010), 29% (2011–2014)). The estimated crude incidence rate of HPV positive OPC increased significantly from 0.62 (1999–2002), 1.06 (2003–2006), 1.52 (2007–2010) to 1.74 (2011–2014) per 100,000 person-year. The sensitivity and specificity of p16 staining for positive HPV infection were 92% and 91%, respectively. The 5-year overall survival rates for patients with HPV positive OPC and with HPV negative OPC were 67.8% and 49.0%, respectively (HR = 0.52 (0.35–0.76), p = 0.0005). Patients with HPV positive OPC but no betel nut/cigarette exposure had the best overall survival (5-year: 88.2%, p < 0.0001). Patients with HPV negative OPC and betel nut/cigarette exposure had the worst overall survival (5-year: 46.6%, p < 0.0001). Patients with HPV positive OPC but also with betel nut/cigarette exposure had poorer 5-year overall survival (48.3%, p < 0.01).ConclusionThe incidence of HPV positive OPC is increasing along with HPV negative OPC, which leads to stably low percentage of HPV positive OPC in Taiwan. HPV positive OPC may become an important head and neck cancer when the incidence of HPV negative OPC declines in the near future. P16 is a useful surrogate marker for HPV infection in OPC and a good prognostic indicator for treatment outcome of OPC. Patients with HPV positive OPC but no betel nut/cigarette exposure has an excellent prognosis. Betel nut/cigarette exposure significantly worsens the prognosis of HPV positive OPC.

Does P16 Protein Expression Affect Treatment Prognosis in Oral Squamous Cell Carcinoma - A Comparative Study.

Introduction:P16 is an independent and reliable surrogate for the detection of human papillomavirus (HPV) in oral squamous cell carcinoma (OSCC). The aim of this study was to assess the P16 expression as a marker for HPV infection in OSCC and its impact on the treatment outcome.Methods and Materials:A cross-sectional study was conducted on patients with a definite diagnosis of OSCC. Patients were assigned into two groups with and without recurrence or metastasis. Tumour resection was performed in the same manner for all patients. P16 expression was evaluated by immunohistochemical staining. Independent t-test and Chi-square tests were used to find significant differences in age, gender, stage of the disease, tumour size, lymph node involvement, and P16 expression between the two groups.Results:Of 50 patients, 37 did not show any recurrence or metastasis (group 1), while 13 had a relapse (group 2). There was no significant difference for age, gender distribution, stage of the disease, or lymph node involvement between the two groups (P > 0.05). A significant difference in tumour size was noted between the two groups (P = 0.001). The mean expression of P16 was 38.92 ± 24.36 in group 1 and 51.54 ± 33.63 in group 2. No significant difference was found between the two groups for the mean expression of P16 (P = 0.23).Discussion:A review of the recent literature revealed that the HPV role in OSCC treatment is controversial. According to the results of this study, there was no significant difference in terms of P16 expression between OSCC patients with and without recurrence or metastasis.

Expression of p16 in cervical premalignant and malignant lesions- IHC study

Cervical cancer is the second leading cause of female cancer in India. Human papilloma virus (HPV) infection is the most important risk factor for uterine cervical neoplasms. HPV oncogene expression can be detected by cellular protein p16 (INK4a) using immunohistochemistry. The present study aims to assess the immunoexpression of protein p16INK4a in cervical dysplastic and neoplastic lesions. The study includes 86 cases of premalignant and malignant cervical lesions diagnosed on histopathology for two years. All cases were subjected to p16 INK4a immunohistochemical staining.72% of cases (62 out of 86) (including premalignant and malignant) were positive for p16. P16 was positive in 88% of invasive carcinoma, 70.83% of HSIL and 40% of LSIL. Cervical cancer is associated with HPV infection which can be identified by overexpression of p16 INK4a in premalignant and malignant lesions. The expression of p16 INK4a positivity increases with increasing grades of dysplasia and thus helps in the stratification of cervical dysplastic and neoplastic lesions. P16 INK4a can be used as a surrogate marker of HPV infection.

Low prevalence of human papillomavirus infection in sinonasal inverted papilloma and oncocytic papilloma.

The aim of this study was to investigate the role of human papillomavirus (HPV) in sinonasal inverted papilloma (SIP) and sinonasal oncocytic papilloma (SOP) from a single institution and whether p16 can serve as a surrogate marker for HPV infection. This study included 49 subjects with SIP and 36 subjects with SOP. Formalin-fixed paraffin-embedded tissues were used to extract genomic DNA, and HPV detection was performed by utilizing a valid nested polymerase chain reaction approach that can detect all known HPV subtypes. Immunohistochemistry was used to evaluate the expression of p16 in all tumor sections. The presence of HPV DNA was found in 6.1% (3/49) of the SIP patients and 11.1% (4/36) of the SOP patients. All identified HPV subtypes in SIP were high-risk HPV, including HPV-16 (two patients) and HPV-58 (one patient). Regarding SOP, there were three patients positive for HPV-16 and one with low-risk HPV (type 6). In total, 11/49 (22.4%) SIP lesions and 10/36 (27.8%) SOP lesions were considered p16 positive, with p16 staining in more than 70% of tumor cells. There was only one SIP and one SOP that were positive for both HPV (high-risk HPV type 16) and p16 staining. HPV does not play an etiologic role in inverted papilloma or oncocytic papilloma of the sinonasal region. p16 immunostaining should not be used as a surrogate marker to evaluate the HPV infection status in these lesions.

The role of protein p16INK4a in non-oropharyngeal head and neck squamous cell carcinoma in Southern China.

Cyclin-dependent kinase inhibitor 2A (p16INK4a) protein is a surrogate immunohistochemical marker of human papillomavirus infection in oropharynx squamous cell carcinoma (OPSCC). However, the effects of p16INK4a in non-OPSCC require additional analysis. In addition, major gaps remain in the literature, including small volumes of data for China. Therefore, the present study evaluated the frequency of p16INK4a positivity in patients with non-OPSCC in Southern China, and assessed its prognostic value. p16INK4a expression status in patients with non-OPSCC was determined by immunohistochemistry. p16INK4a-positive expression was defined as a strong and diffuse staining in ≥70% of the tumor cells. Then, the diagnostic value of p16INK4a in predicting overall survival (OS) and disease-free survival (DFS) rate was determined. The positive rate of p16INK4a was 26.3% in larynx cancer and 24.8% in oral cavity cancer. Multivariate analysis revealed that the protein status independently predicted improved OS rate, but not DFS rate (P=0.096). Comparing different disease stages, patients at an early stage with p16INK4a-positive non-OPSCC exhibited improved DFS and OS rates compared with those exhibited by patients who were negative. The p16INK4a-positive rate in patients with non-OPSCC was 25.1% [26.3% in Laryngeal squamous cell carcinoma (LSCC) and 24.8% in Oropharyngeal squamous cell carcinomas (OSCC)] in the present cohort from South China. The present study suggested that p16INK4a expression in non-OPSCC predicts favorable clinical outcomes, particularly in early stage non-OPSCC and oral cancer.

Clinico-pathological features of oropharyngeal squamous cell carcinomas in Malaysia with reference to HPV infection

BackgroundThe incidence of oropharyngeal squamous cell carcinoma (OPSCC) has been rising in Western countries and this has been attributed to human papillomavirus (HPV) infection. p16 expression is a marker for HPV infection and p16 positive OPSCC is now recognized as a separate disease entity. There are only limited data available regarding HPV-related OPSCC in Asian countries and no data from Malaysia.MethodsWe identified 60 Malaysian patients with OPSCC over a 12-year period (2004–2015) from four different hospitals in two major cities, Kuala Lumpur and Penang. The detection of HPV was carried out using p16 immunohistochemistry and high risk HPV DNA in situ hybridisation.ResultsOverall, 15 (25%) tumours were p16 positive by immunohistochemistry, 10 of which were also positive for high risk HPV DNA by in situ hybridisation. By comparison, a matched cohort of UK patients had a p16 positive rate of 49%. However, between 2009 and 2015, where cases were available from all four hospitals, 13 of 37 (35%) cases were p16 positive. In our Malaysian cohort, 53% of patients were of Chinese ethnicity and 80% of the p16 positive cases were found in these patients; no Indian patients had p16 positive disease, despite representing 35% of the total cohort.ConclusionThe proportion of OPSCCs associated with HPV in Malaysia appears to be lower than in European and American cohorts and could possibly be more prevalent amongst Malaysians of Chinese ethnicity. Further, our data suggests that the burden of HPV-related OPSCC could be increasing in Malaysia. Larger cross-sectional studies of Malaysian patients are required to determine the public health implications of these preliminary findings.

Expression of cell cycle-related proteins in oropharyngeal squamous cell carcinoma based on human papilloma virus status.

The molecular association between human papilloma virus (HPV) and cell cycle-related (CCR) proteins is not fully understood in oropharyngeal squamous cell carcinoma (OPSCC). Herein, we examined the expression levels of CCRs in OPSCCs based on HPV status. Insitu hybridization (ISH)- and polymerase chain reaction (PCR)-based assays were used to detect HPV status in 98OPSCCs, and CCRs were detected by immunostaining. Of 98tumors, 47had HPV-positive tumors by either ISH- or PCR-based assays. Concordance analysis between ISH- and PCR-based tests showed a good agreement in OPSCC. Expression of p16 was moderate, consistent with HPV positivity as determined by ISH-based analysis. High p53-, phospho (p)-Rb-, SKP2-, cyclinD1-, and p-c-myc-positive rates were correlated with HPV-negative tumors, whereas high p16- and p27-positive rates were associated with HPV-positive tumors. The positive rates for p21 and cyclinA did not differ between HPV-positive and HPV-negative tumors. Finally, Ki-67 positivity was commonly observed for both types of OPSCCs. Although expression of p16 is thought to be a marker of HPV infection, ISH- or PCR-based tests should be used for HPV detection. In addition, our results regarding CCRs may be helpful for understanding the carcinogenesis of OPSCC.

P16INK4a is not a Reliable Screening Marker of HPV Infection in Esophageal Squamous Cell Carcinoma: Evidence from a Meta-Analysis

The role of p16INK4a as a surrogate marker for screening human papillomavirus (HPV) in esophageal squamous cell carcinoma (ESCC) remains controversial. A comprehensive search of EMBASE, PubMed, China National Knowledge Infrastructure and China Biology Medicine was performed from inception to December 27, 2015. A random-effects model was applied to the pooled odds ratios (ORs) with 95% confidence intervals (CIs). Ten studies were identified (985 cases). The pooled results showed no significant relationship between p16INK4a expression and HPV infection in ESCC based on overall HPV types (OR: 1.79, 95% CI: 0.69-4.66, p = 0.235). Subgroup analysis by HPV detection method showed no statistical significance in either the polymerase chain reaction (PCR) (OR: 1.65, 95% CI: 0.83-3.30, p = 0.154) or in situ hybridization (ISH) group (OR: 2.58, 95% CI: 0.03-268.14, p = 0.689). The pooled OR of the sensitivity analysis ranged from 1.27 (95% CI: 0.58-2.84) to 2.32 (95% CI: 0.95-5.64). Of these studies, 6 involved only high-risk human papillomavirus types (HR-HPV), HPV16 or HPV18. However, similar observations were made for HR-HPV (OR = 1.31, 95% CI: 0.26-6.59, p = 0.741). Subgroup analysis again showed no statistical significance in the PCR group (OR: 0.95, 95% CI: 0.25-3.64, p = 0.940) and ISH group (OR: 2.58, 95% CI: 0.03-268.14, p = 0.689). Sensitivity analysis showed that the pooled OR ranged from 0.69 (95% CI: 0.21-2.22) to 1.89 (95% CI: 0.33-10.86). p16INK4a is not a reliable screening marker of HPV infection in ESCC. Further multicenter, large-sample and well-matched prospective studies are still required to illuminate the possible etiological roles of HPV in ESCC.

Abstract 1661: TRIP12 as a mediator of human papillomavirus/p16-related radiation enhancement effects

Abstract Objective: Patients with human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) have better responses to radiotherapy and higher overall survival rates than do patients with HPV-negative HNSCC, but the mechanisms underlying this phenomenon are unknown. P16 is used as a surrogate marker for HPV infection. Our goal was to examine the role of p16 in HPV-related favorable treatment outcomes and to investigate the mechanisms by which p16 may regulate radiosensitivity. Methods: HNSCC cells and xenografts were used. P16-overexpressing (HPV/p16-negative HN5 and UMSCC-1) and p16 shRNA knockdown (HPV/p16-positive UMSCC-47 and UCPI-SCC154) cells, TRIP12 shRNA or siRNA knockdown (HPV/p16-negative HN5 and Fadu) cells were generated. The effects of p16 or TRIP12 on HNSCC cell radiosensitivity were determined by clonogenic cell survival. The effects of p16 on tumor xenografts (HPV/p16-negative HN5 and HPV/p16-positive UMSCC-47) radioresponse were evaluated by tumor growth delay assays. DNA double-strand breaks (DSBs) were assessed by immunofluorescence analysis of 53BP1 foci; DSB levels were determined by neutral comet assay; western blotting was used to evaluate protein changes; changes in protein half-life were tested with a cycloheximide assay; gene expression was examined by real-time polymerase chain reaction (PCR); and data from The Cancer Genome Atlas HNSCC project were analyzed. Results: P16 expression led to downregulation of TRIP12 protein both in vitro and in vivo via decreasing TRIP12 protein's half life, which in turn led to increased RNF168 levels and subsequently repressed DNA damage repair, represented by increased 53BP1 foci and neutral comet moment tails at 24 hours after irradiation. As a result, p16 expression enhanced radioresponsiveness both in vitro and in vivo. Inhibition of TRIP12 expression led to radiosensitization through repressed DNA damage repair, represented by decreased BRCA1 foci at 1 and 5 hours after irradiation; and increased neutral comet moment tails at 24 hours after irradiation. Furthermore, overexpression of TRIP12 was associated with poor survival in patients with HPV-positive HNSCC. Conclusions: The findings of our study reveal that p16 participates in radiosensitization through influencing DNA damage repair. P16 downregulates TRIP12 protein expression via post-translational regulation. Inhibition of TRIP12 sensitized HNSCC cells via prohibition of DNA damage repair. These findings support the rationale of blocking TRIP12 signaling to improve radiotherapy outcomes. Citation Format: Li Wang, Peijing Zhang, David Molkentine, Jessica Molkentine, Uma Raju, David Valdecanas, Ramesh Tailor, Howard Thames, Thomas Buchholz, Junjie Chen, Li Ma, Kathy Mason, Raymond Meyn, Heath D. Skinner. TRIP12 as a mediator of human papillomavirus/p16-related radiation enhancement effects. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1661.

Detection of human papillomavirus infection in oral squamous cell carcinoma: a cohort study of Japanese patients.

Recent evidence suggests that human papillomavirus (HPV)-related head and neck squamous cell carcinoma (HNSCC) is a separate HNSCC subgroup with distinct epidemiology, histopathological characteristics, therapeutic response to chemotherapy and radiation, and clinical outcome. This study aimed to investigate the role of HPV infection in oral squamous cell carcinoma (OSCC) and the correlation between HPV infection, tumor suppressor protein p16 expression, and clinicopathological features in Japanese patients. In total, 174 OSCC specimens were examined for p16 levels by immunohistochemistry, and p16-positive OSCCs were analyzed for HPV DNA by in situ hybridization (ISH) and HPV genotypes by real-time PCR. The results were evaluated for the association with clinicopathological characteristics of OSCC patients. Twenty-four OSCC samples were found positive for p16 expression; all of them were well-differentiated tumors. P16 immunoreactivity was significantly associated with the invasion depth and tended to correlate with sex, site in the oral cavity, stromal reaction, TNM stage, and survival. HPV DNA was detected in 13 of 24 (54%) p16-positive OSCC by real-time PCR; HPV 16, 18, and other high-risk genotypes were the most prevalent. However, ISH failed to detect HPV DNA in p16-positive OSCCs. P16 immunoreactivity and HPV genotyping by real-time PCR may be useful markers of HPV infection in OSCC. However, although HPV-related OSCC showed good outcomes, HPV infection may have a minor role in oral oncogenesis in Japanese patients.

Human papillomavirus infection in Bowen disease: negative p53 expression, not p16(INK4a) overexpression, is correlated with human papillomavirus-associated Bowen disease.

Genital Bowen disease (BD) has been linked to the high-risk types of human papillomavirus (HPV) infection. Recently, it has been recognized that HPV also can be associated with extragenital BD. HPV oncoproteins E6 and E7 interfere with the function of p53 and pRb, respectively, leading carcinogenesis. p16(INK4a) overexpression induced by inactivation of pRb is recognized as a surrogate marker for HPV-associated cervical cancer. In this study, we examined the presence of HPV DNA in 142 BD lesions by polymerase chain reaction (PCR), and determined the type of HPV by PCR restriction fragment length polymorphism or direct DNA sequencing. HPV DNA was detected in 66.7% of genital BD and 8.3% of extragenital BD. The types of HPV detected were HPV types 6, 16, 33, 52, 56, 58 and 59. We also investigated the expression of p16(INK4a) , pRb and p53 by immunohistochemistry. Positive expression was detected in 88.6% for p16(INK4a) , 25.2% for pRb, and 63.8% for p53. There was no significant difference in p16(INK4a) and pRb expression between HPV-positive and -negative BD. However, a strong correlation of HPV positivity with p53 negativity was found. A total of 66.7% of HPV-positive BD showed no p53 expression, whereas the corresponding rate was 32.8% of HPV-negative BD. This study demonstrated that HPV can participate in the development of BD, not only in the genital lesion, but also in extragenital lesion. p16(INK) (4a) overexpression is not a marker for HPV infection in BD. Instead, negative p53 expression is correlated with HPV-associated BD.

Immunohistochemical expression of p16 and HPV L1 capsid proteins as predictive markers in cervical lesions

Human papilloma virus (HPV) infection is the most important cause of cervical cancer, but only 2 % of cervical HPV infections will develop into cervical cancer. p16 INK4A has been introduced as a marker for HPV infection in cervix. HPV L1 capsid protein is also known to be associated with the productive phase of HPV infection; however, expression pattern in different HPV-associated cervical lesion and its correlation to p16 expression is not still well understood. The authors aimed to elucidate the relationship between L1 and p16 expression in cervical lesions. Immunohistochemical studies using antibodies against L1 capsid and P16 proteins were carried out on 89 paraffin-embedded tissue samples including 11 low-grade cervical intraepithelial neoplasias (CIN), 11 high-grade CINs, 20 cervical squamous cell carcinomas (SCC), eight cervical adenocarcinomas and 39 normal cervical tissues as a control group. L1 capsid protein was positive in 63.6 % of low-grade CINs and 9.1 % of high-grade CINs; while none of the cervical SCCs, adenocarcinomas or normal cervical tissues showed this marker. In contrast, p16 protein was positive in 81.8 % of low-grade CINs, 90.1 % of high-grade CINs, 90 % of SCCs, 75 % of adenocarcinomas and 10.25 % of normal cervical tissues (p value < 0.001). Despite the presence of interobserver variation in the histopathologic interpretation of cervical lesions, in more instances definite diagnosis is made by routine histopathological examination and these ancillary tests are supportive in follow-up of the patient.

P16 expression as a human papillomavirus (HPV)-independent prognostic biomarker in non-oropharyngeal squamous cell carcinoma (non-OPSCC).

6007 Background: p16 is an important tumor suppressor and cell cycle regulator that is commonly lost in HPV-negative (-) and upregulated in HPV-positive (+) OPSCC. While the role of p16 expression in OPSCC as a surrogate marker of HPV infection and its association with prognosis are well established, its significance has not been studied in non-OPSCC including oral cavity, hypopharynx and larynx, where HPV infection is rare. We hypothesize that p16 expression is a prognostic biomarker of favorable outcome (progression-free and overall survival) in non-OPSCC. Methods: p16 expression in non-OPSCC from RTOG 0129, 0234 and 0522 was determined by immunohistochemistry (p16 mouse monoclonal antibody, prediluted, MTM-CINtech, 9518) and considered positive if &gt;70% of the tumor cells demonstrated diffuse staining. The high risk HPV status in non-OPSCC from RTOG 0129 and 0522 was determined by in situ hybridization using a cocktail probe including HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 66. Hazard ratios from Cox models were expressed as positive/negative, stratified by trial, and adjusted for age, gender, T, and N stage. Results: p16 expression was positive in 14.1% (12/85), 24.2% (23/95) and 19.0% (27/142) of non-OPSCC from RTOG 0129, RTOG 0234, and RTOG 0522, respectively. HPV ISH was positive in 6.5% (6/93) and 6.9% (7/101) of non-OPSCC from RTOG 0129 and RTOG 0522, respectively. There was moderate agreement between p16 and HPV status (kappa=0.53). The hazard ratios for p16 expression are 0.63 (p=0.03) and 0.56 (p=0.01) for PFS and OS, respectively. In 0522, there was no interaction between p16 status and two treatment arms (RT+cisplatin+/-cetuximab). In comparison of OPSCC and non-OPSCC, patients with p16(+) OPSCC have better PFS and OS than patients with p16(+) non-OPSCC, but patients with p16(-) OPSCC and non-OPSCC have similar outcomes. Conclusions: Similar to results in patients with OPSCC, patients with p16(+) non-OPSCC have better outcomes than patients with p16(-); however, the differential appears smaller than has been observed in HPV(+) OPSCC. Further studies are warranted to delineate the role of p16 expression as a prognostic biomarker in non-OPSCC and OPSCC. Clinical trial information: NCT00265941, NCT00047008, NCT00084318.

Markers of HPV infection and survival in patients with head and neck tumors

The purpose of this study was to determine whether changes in human papillomavirus (HPV) DNA prevalence in oral rinses and/or HPV-specific antibody levels in the sera of patients with oral/oropharyngeal cancer have prognostic significance. One hundred and forty-two patients with oral/oropharyngeal tumors were enrolled. The presence of HPV DNA was assayed in tumor tissue and oral rinses and HPV-specific antibodies were assessed in the sera. Oral rinses were collected before treatment and one year after the treatment. Sera were drawn before treatment, one month, and one year after the end of the treatment. Altogether, 59.2% of tumors were HPV positive. The presence of HPV DNA in the tumors correlated with HPV DNA positivity in oral rinses and with HPV-specific antibodies in the sera. Out of 66 patients with HPV-positive oral rinses at enrolment, 84.8% became negative at one-year follow-up, while most patients remained seropositive for HPV-specific antigens. However, the mean titers of HPV16 E6 and/or E7 antibodies at follow-up were significantly lower. Of 16 patients with recurrences at follow-up (alive on second sampling), six were positive at enrolment for HPV16 E6 and/or E7 antibodies. In five of these, no decrease in antibody levels was observed. Titers of antibodies specific for HPV16 capsid antigens did not change during the follow-up. Our data suggest that the detection of antibodies specific for the HPV 16 E6 and E7 oncoproteins may serve not only as a marker of HPV etiology, but also as a marker of recurrence and a prognostic indicator in patients with HPV-positive tumors.

Abstract A12: Web-based tools for visualization of the Human Papilloma Virus genome dynamics: A computational approach to cancer prevention and control

Abstract The implementation and widespread use of prophylactic Human Papilloma Virus (HPV) vaccines targeting the most common oncogenic HPV types (HPV 16/18), is expected to change the prevalence of high risk types in populations with high vaccination rates. The projected loss in the Positive Predictive Value (PPV) of cytology in the post vaccination era suggests a need to rely on molecular markers of HPV infection and technologies, such as HPV genotyping. FDA approval of HPV genotyping tests has led to concerns about how typing can assist cervical cancer screening and clinical decisions in a cost-effective manner. Computational tools to visualize the dynamics of genetic material across related HPV genomes are lacking. Web-and Mobile-based tools, which make sequencing results easily accessible to clinicians, investigators, and patients, can be created to facilitate functional interpretations, spur biomarker development, and advance the deployment of medical devices for cancer prevention and control. To facilitate the visualization and functional interpretation of high risk genotypes in hpv-associated cancer patients we downloaded 21 HPV genomes from the NCBI public repository (http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Tree&amp;id=151340&amp;lvl=3&amp;lin=f&amp;keep=1&amp;srchmode=1&amp;unlock) and created two proof-of-principle Web-based tools, which allow the comparative view of 11 high risk HPV types and 9 low risk HPV types against a reference genome (hpv16). All the 20 genomes play a role in HPV-associated benign or malignant lesions. The Enterix-HPV web servers (http://enterix.cbcb.umd.edu/enteric/enteric-hpv.html), (http://enterix.cbcb.umd.edu/menteric/menteric-hpv.html) allow comparative views of twenty human papilloma virus (HPV) genomes (hpv18, hpv45, hpv68b, hpv53, hpv31, hpv52, hpv6, hpv54, hpv33, hpv56, hpv11, hpv61, hpv35, hpv58, hpv42, hpv72, hpv39, hpv59, hpv44, hpv81) against a reference genome (hpv16), in two formats: large-scale and close-up. The large-scale view (Enteric) displays pair wise alignments and annotations of genome rearrangement events in a 9KB region surrounding an address in the reference genome, whereas the close-up view (Menteric) shows nucleotide-level multiple alignments of the genomes and annotations of conserved and functional regions in a 1KB region. Hyperlinks embedded in the graphical features link to web resources such as the Entrez Gene pages, provide additional information about the feature, or allow downloading sequence data for local analyses. It is anticipated that screening for oncogenic HPV genotypes, followed by cytological triage, may be useful in a post-vaccination era. Genotyping for specific oncogenic HPV types has been shown to increase the PPV and specificity in predicting CIN2/3 and carcinoma in women with ASCUS and LSIL, thus possibly tailoring clinical management of these women in a post vaccination era. Enterix-HPV can be a useful tool for the new wave of Next generation sequencing-based approaches to HPV genotyping for cervical cancer clinical management, prevention and control. Computational tools, such as Enterix-HPV, that create Web- and Mobile-based visualization solutions can be used to spur innovation in HPV-related cancer prevention and clinical care. As HPV genotyping becomes the standard of care in screening and clinical management of HPV related tumors, interactive Web- and Mobile-based genomic visualization tools will become valuable assets for clinicians, research scientists, and patients. Citation Format: Rafael Guerrero-Preston, Michael McClelland, David Sidransky, Liliana Florea. Web-based tools for visualization of the human papillomavirus genome dynamics: A computational approach to cancer prevention and control. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A12.

Clinical and Pathologic Factors of Prognostic Significance in Penile Squamous Cell Carcinoma in a North American Population

To analyze, in a clinicopathologic correlation study, a small population of primarily white men with invasive squamous cell carcinoma of the penis for potential prognostic predictors. Penile squamous cell carcinoma is an uncommon cancer in North America. It has a wide spectrum of clinical behavior and an understudied pathogenesis. The data from 43 patients with invasive squamous cell carcinoma of the penis were studied retrospectively. Extensive chart reviews were conducted, glass slides were reviewed, and tissue microarrays were constructed for analysis of immunohistochemical stains p16(INK4a), p53, and Ki-67. Univariate and multivariate logistic regression analyses were performed to elucidate any clinical or pathologic factors that would predict overall survival. The mean age at diagnosis was 63 years. Most cases (63%) were invasive squamous cell carcinoma, not otherwise specified, and presented as pathologic stage T1 or T2 tumors. Of the 43 patients, 23% died of their disease; 53% of the cases stained for p16(INK4a). Higher pathologic tumor stage and a lack of p16(INK4a) staining were independent predictors of worse overall survival (P = .014) and cancer-specific survival (P = .010). Our results have shown that 53% of the invasive penile squamous cell carcinoma cases in this population were associated with human papillomavirus, using p16(INK4a) as a surrogate marker of human papillomavirus infection. These patients had a statistically significant survival advantage, independent of other prognostic factors.