HPV-positive Squamous Cell Carcinoma Research Articles

Assessment of Patient Preferences among the Possible HPV-positive Squamous Cell Carcinoma Population

Assessment of Patient Preferences among the Possible HPV-positive Squamous Cell Carcinoma Population

Association of human papillomavirus and squamous cell carcinoma of esophagus: A SEER database analysis.

e14634 Background: Squamous cell carcinoma (SCC) of the esophagus is generally associated with tobacco and alcohol use. However, there are multiple published reports suggesting a subset of SCC of esophagus is associated with human papillomavirus (HPV) infection. HPV is well known to be associated with SCC of the anus and tonsil. Patients with HPV-positive SCC do well with primary chemoradiation and it is possible subset of esophageal cancer might also do well with primary chemoradiation. Prior analyses of the Surveillance, Epidemiology, and End Results (SEER) database showed that the incidence of these HPV associated SCC in the younger population are increasing. We used the SEER database (1973-2007) to evaluate epidemiological similarities between HPV associated SCCs. Methods: We assessed the association of HPV and SCC of esophagus by evaluating epidemiological similarities between SCC of tonsil, anus, and esophagus among the younger age group (age 20 to 44) and older age group (age above 45) of patients. Patient characteristics, 5-year relative survival, and annual percentage changes (APC) in incidence rates were evaluated. Results: We identified 15,825 SCC of the tonsil, 25,943 SCC of the esophagus, and 9,437 anal SCC. Among the younger age group, a significant increase in the APC was seen in anal SCC (APC= +6.7; 95% CI [5.3,8.0]; p<0.05) and SCC of tonsil (APC= +3.5; 95% CI [2.5,4.5]; p<0.05), whereas the APC of SCC of esophagus in younger age group showed a significant decrease (APC= -2.0; 95% CI [-2.8, -1.1]; p<0.05) (Table). Conclusions: Despite multiple studies suggests SCC of esophagus is associated with HPV infection, the incidence of SCC of the esophagus has not changed in the same manner as SCCs that are strongly associated with HPV. This suggests that in the population studied, SCC of the esophagus may not be associated with HPV. Future work includes identifying subgroups of the patients whose SCC was probably associated with HPV by incorporating factors such as race or sex. Age 20-44 Age ≥ 45 5-year relative survival APC 95% CI 5-year relative survival APC 95% CI Anal SCC 67.2% 6.7* 5.3, 8.0 68.3% 3.5* 3.1, 3.9 Tonsil SCC 70.1% 3.5* 2.5, 4.5 54.1% 0.3* 0.1, 0.6 Esophageal SCC 18.6% -2* -2.8, -1.1 10.4% -2.4* -2.9, -2.0 * p<0.05

Panitumumab treatment of head and neck cancer after cetuximab-induced aseptic meningitis: Case report.

e16033 Background: Aseptic meningitis is a rare complication of the initial, loading-dose infusion of cetuximab, with 4 reported cases (2 lung cancer, 2 head/neck cancer.) Re-treatment with cetuximab at standard, lower weekly dosage with corticosteroid preparation was possible in 2 cases. Methods: I report a 54 y.o. man with risperidone-controlled Tourette syndrome, who presented for initial loading dose of cetuximab for stage IV-A, HPV-positive squamous cell carcinoma of left base of tongue, as part of definitive IMRT radiation treatment plan. Hydrocortisone was added to ranitidine-diphenhydramine premedication to lessen chances of adverse neurological reaction. He received standard cetuximab loading dosage of 400 mg/m2 by 90-minute intravenous (i.v.) infusion. Results: The initial test and loading infusions of cetuximab were uneventful. Sixteen hours later, he developed headache, nausea, fevers, photophobia, and blurred vision. Imaging by CT and MRI of brain revealed no hemorrhage, edema, leptomeningeal enhancement, or metastases. Lumbar puncture yielded clear, colorless fluid with 661 nucleated cells/µl (97% neutrophils), no red blood cells, glucose 94 mg/dl, and protein 74 mg/dl. He received empiric antibiotics and antiviral therapy for 3 days. Cultures of spinal fluid and blood were negative. He defervesced and symptoms returned to baseline in 2 days. Radiation therapy was initiated 5 days later, and he received i.v. panitumumab, 3 mg/kg/week, with the same premedications, as adjuvant beginning on day 1 of definitive radiation. There were no further neurological, anaphylactic, or allergic events during therapy. He did develop typical eczematoid skin rash during therapy. Response status is pending. Conclusions: Panitumumab was an effective, safe adjuvant systemic therapy in this man with severe neurological reaction to cetuximab. This agent is currently under investigation as adjuvant with definitive radiation therapy for head and neck cancers, and appears to be a safe alternative to cetuximab in this setting. Possible mechanisms for this phenomenon will be presented.

Prognosis of HPV-positive squamous cell carcinoma of tonsil-expressing high level of p16 and low level of EGFR.

5546 Background: HPV infection has been known to be one of the important risk factors for carcinogenesis of head and neck squamous cell carcinoma (HNSCC). The over-expression of epidermal growth factor receptor (EGFR) correlates with a poor prognosis in HNSCC. We investigated the relationship between HPV status and EGFR expression and the prognostic impact of these markers in tonsil cancer. Methods: Eligibility criteria was the patients diagnosed as SCC of tonsil and received curative radical resection with available tumor tissue samples. In situ hybridization for HPV and immunohistochemistry for p16, EGFR, and p53 were performed. Results: Among 66 tonsil cancer patients, 24 (36.3%) patients showed HPV positivity. There were no statistically significant differences in clinical characteristics including sex, age, smoking history, TNM stage and treatment modalities between HPV-positive and -negative patients. The over-expression and high intensity of p16 had significant correlation with HPV positivity (p=0.003, 0.017). However, no correlation was found between p53 and HPV infection. There was a trend that the proportion of EGFR over-expression was greater in HPV-negative than in HPV-positive patients (p=0.082), and no or low EGFR intensity was observed in the patients showing p16 over-expression (p=0.015). Among the HPV-positive patients, the frequency of EGFR over-expression was significantly higher in ex- or current smokers (p=0.005). There was meaningful difference of three years disease-free survival between never smokers showing both over-expression of p16 and low expression of EGFR and the others (90.0% and 73.5%, respectively, p=0.082). On univariate analysis, the significant clinicopathologic factors affecting disease-free survival were smoking history and EGFR intensity (p=0.020 and 0.017, respectively). Conclusions: Taken together, p16 was a good surrogate marker for HPV infection. The expression of EGFR was inversely correlated with HPV positivity, but HPV-positive patients with smoking history showed EGFR over-expression. In the HPV-positive tonsil cancers, high p16 and low EGFR expression should be considered as favorable prognostic markers. No significant financial relationships to disclose.

Three synchronous HPV‐associated squamous cell carcinomas of Waldeyer's ring: Case report and comparison with Slaughter's model of field cancerization

Patients with squamous cell carcinoma (SCC) of the oropharynx have an 8% to 20% risk of a synchronous or metachronous second malignancy. The rate of synchronous lesions in human papillomavirus (HPV)-positive oropharyngeal cancers is unknown. We report the case of a 46-year-old man with 3 simultaneous primary malignancies of Waldeyer's ring: HPV-positive SCC of both tonsils and the nasopharynx, with bilateral neck metastases. The patient received induction chemotherapy followed by definitive radiotherapy and remains free of disease at 18 months. Comparison with the Slaughter model of field cancerization suggests that HPV-positive SCC of the head and neck may have a distinct mechanism for the development of multifocal disease. Therefore, the emerging population of young patients with head and neck SCC with HPV-positive cancers presents a new opportunity for understanding the molecular origins of synchronous tumors.

Human Papillomavirus Gene Expression in Cutaneous Squamous Cell Carcinomas from Immunosuppressed and Immunocompetent Individuals

Epidermodysplasia verruciformis (EV)-type human papillomavirus (HPV) DNA have been detected by PCR in squamous cell carcinomas (SCC) from both organ transplant recipients (OTR) and immunocompetent individuals. Their role in skin cancer remains unclear, and previous studies have not addressed whether the viruses are transcriptionally active. We have used in situ hybridization to investigate the transcriptional activity and DNA localization of HPV. EV-HPV gene transcripts were demonstrated in four of 11 (36%) OTR SCC, one of two (50%) IC SCC, and one of five (20%) OTR warts positive by PCR. Viral DNA co-localized with E2/E4 early region gene transcripts in the middle or upper epidermal layers. Non-EV cutaneous HPV gene transcripts were demonstrated in one of five (20%) OTR SCC and four of 10 (40%) OTR warts. In mixed infections transcripts for both types were detected in two of six (33%) cases. Our results provide evidence of EV-HPV gene expression in SCC; although only a proportion of tumors were positive, the similarly low transcriptional activity in warts suggests this is an underestimate. These observations, together with emerging epidemiological and functional data, provide further reason to focus on the contribution of EV-HPV types to the pathogenesis of cutaneous SCC.

Persistence of human papillomavirus infection as a predictor for recurrence in carcinoma of the cervix after radiotherapy

This study was undertaken to examine a possible correlation between clearance or persistence of human papillomavirus (HPV) infection and radiation response in carcinoma of the cervix. We reviewed 97 patients with HPV-positive cervical squamous cell carcinoma (International Federation of Gynecology and Obstetrics [FIGO] stage IB-IVA) treated with radiotherapy. Examination of HPV DNA was performed by the polymerase chain reaction-based assay. Cervical HPV DNA cleared in 42 patients (43.3%) and persisted in 55 patients (56.7%) at the end of irradiation. All of 42 HPV-cleared patients (100.0%) and 50 of 55 HPV-persistent patients (90.9%) had complete response. Of 92 patients with complete response, 20 (21.7%) had local recurrence develop. The recurrence rate was significantly higher in HPV-persisted patients (34.0%) than in HPV-cleared patients (7.1%) ( P = .0016). Univariate analysis demonstrated the significant differences for both 5-year local disease-free survival (LDFS) and overall survival (OAS) between HPV-cleared and HPV-persisted groups. Multivariate analysis showed that persistence of HPV DNA was the most powerful independent predictor for LDFS and OAS compared with other prognostic factors, such as FIGO stage or node swelling. In HPV-positive cervical carcinoma, persistence of HPV DNA in the cervix at the end of irradiation was highly predictive of LDFS and OAS.

Methylation of the Putative Tumor Suppressor Gene RASSF1A in Primary Cervical Tumors

The methylation level of 13 CpG-dinucleotides in the promoter region of the putative tumor suppressor gene RASSF1A (3p21.31) was analyzed in HPV-positive squamous cell carcinomas of cervix using methyl-sensitive restriction endonuclease analysis followed by PCR. The methylation from 3 to 13 CpG-dinucleotides was observed in 64% (25/39) tumors, 22% (2/9) morphologically normal tissues adjacent to tumors (P = 0.0306) and in 2 from 3 leucocytes of peripheral blood of patients. The methylation of these CpG-dinucleotides was absent in DNA of healthy donor leucocytes (0/10). Methylation level of the examined fragment of the RASSF1A promoter region was significantly higher in tumors of patients with lymph node metastases in comparison to tumors of patients without metastases (P = 8.5 x 10(-12)). The methylation frequency of RASSF1A gene was in two times higher than hemi- and homozygous deletion frequency at the region of location of this gene (chromosome 3p21.31), determined earlier. These data suggest that methylation of the RASSF1A gene is one of the main ways of this gene inactivation in HPV-positive cervical squamous cell carcinomas. The methylation of the RASSF1A gene is an early event in genesis of tumor and the level of methylation increased with tumor progression.

Loss of caspase-8 activation pathway is a possible mechanism for CDDP resistance in human laryngeal squamous cell carcinoma, HEp-2 cells

Cisplatin (CDDP) is among the most widely used and most effective chemotherapeutic agent for many types of human cancer. Because killing cancer cells by chemotherapy is principally executed by apoptosis, a defective apoptotic program might acquire drug resistance. Flow cytometric Annexin V assay demonstrated that HEp-2 cells (human laryngeal cancer) were persistently resistant to CDDP as compared to HeLa cells (human uterine cervical cancer), despite the same histological type and wild-type p53 status. CDDP treatment caused steady induction of p53 protein in both cancer cell types, although it was more dramatic in CDDP-resistant HEp-2 cells, which was correlated well with p53 Ser15 phosphorylation, but not with the expression level of HPV type 18 E6 oncoprotein in these cells. Importantly, CDDP differently activated caspase cascades between HEp-2 and HeLa cells. CDDP activated the caspase-8 pathway through TNFR superfamily receptors such as Fas, but not caspase-9 in HeLa cells. On the other hand, the caspase-9 pathway was significantly activated in HEp-2 cells, although the activation of caspase-8 by CDDP was deficient. This different response to CDDP in caspase-8 activation was not related with the expression level of either Fas or FasL in these cells. We concluded from these results that loss of the caspase-8 activation pathway in HEp-2 cells was a possible mechanism for its resistance to CDDP-induced apoptosis. The caspase-8 pathway might play an important role in CDDP-induced apoptosis in HPV-positive human squamous cell carcinomas.

The expression of key cell cycle markers and presence of human papillomavirus in squamous cell carcinoma of the tonsil.

Chemical carcinogens induce squamous cell carcinoma (SCC) of the head and neck by targeting the p53 and the retinoblastoma (pRb) pathways. Human papillomavirus (HPV) might have an etiologic role in these cancers at particular sites. Few studies have compared cell cycle protein expression in HPV-positive and HPV-negative tumors in this region. Fifty tonsil SCCs were analyzed for HPV by PCR and for expression of cell cycle proteins (p53, pRb, p16(INK4A), p21(CIP1/WAF1), p27(KIP1), and cyclinD1) by immunohistochemistry. HPV was present in 42%; almost all were type 16. There were statistical associations between HPV positivity and reduced expression of pRb and cyclinD1, overexpression of p16, and younger patient age. Tumor with down-regulated p27 tended to have down-regulated pRb and p21. HPV-positive tonsil SCCs have distinct molecular pathways. Their association with younger patient age suggests that they are biologically distinct from HPV-negative tumors.

Expression of p16, Rb, and p53 Proteins in Squamous Cell Carcinomas of the Anorectal Region Harboring Human Papillomavirus DNA

Human papillomavirus (HPV) has been implicated as an etiologic agent for the development of squamous cell carcinoma of the anorectal region. It has been shown that the HPV E6 and E7 oncoproteins are able to inactivate the tumor suppressor functions of p53 and Rb. In cervical and head and neck cancers, HPV infection is also associated with an overexpression of p16, a cyclin-dependent kinase inhibitor. The expression of these cell cycle regulators in squamous cell carcinomas of the anorectal region has not been well studied. In the current study, 29 cases of squamous cell carcinoma of the anorectal region were immunohistochemically examined for the expression of p16, Rb, and p53 proteins. Tumor cell DNA was also extracted from paraffin blocks and subjected to broad-spectrum HPV DNA testing and typing. The results show that the tumor cells exhibited a strong and diffuse nuclear stain (with some cytoplasmic positivity) for p16 in all 29 cases (100%). The adjacent nonneoplastic squamous epithelium or colonic mucosa, in contrast, was completely negative. Loss of Rb nuclear staining in tumor cells was observed in 20 cases (69%). The p53 protein was essentially undetectable, with only 6 cases containing <10% positive cells. HPV DNA was detected in every case (100%), with 25 cases (86%) harboring Type 16. In addition, almost identical results were obtained in 12 HPV-positive squamous cell carcinomas of the upper aerodigestive tract. This was in marked contrast to those of HPV-negative tumors, where positive p16 staining and loss of Rb expression were seen in only 2/21 (10%) and 1/21 (5%) cases, respectively. These observations indicate that overexpression of p16 and loss of Rb nuclear staining are commonly associated with high-risk HPV infection, which may serve as useful surrogate biomarkers for identifying squamous cell carcinomas harboring HPV DNA.

Expression of p21/WAF‐1, status of apoptosis and p53 mutation in esophageal squamous cell carcinoma with HPV infection

Human papilloma virus (HPV) is regarded as a causative carcinogenic agent in anogenital squamous cell carcinoma (SCC), but there is controversy about its etiologic role in esophageal SCC (ESCC). In this study, we attempted to clarify whether HPV infection plays a crucial role in the development of ESCC by analysis of multiple factors. These included: detection of HPV DNA; evaluation of immunohistochemical assays for HPV-related cell cycle regulators and apoptosis by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method; and genetic analysis of the p53 gene. Twenty of the 48 ESCC examined (42%) were found to be positive for the HPV genome by polymerase chain reaction. They comprised 16 cases with the HPV16 subtype, three with the HPV18 subtype, and one with both HPV16 and 18. Immunohistochemical analysis revealed that the expression of p21/WAF-1 was significantly decreased in HPV-positive cases (chi2 = 9.2614; P = 0.0023). Furthermore, the 10 apoptosis-negative (< or =10%) cases of HPV-positive SCC were almost exclusively p21/WAF-1-negative (chi2 = 12.1406; P = 0.0005), indicating the significance of the relationship between HPV infection and the phenotype that is expected from HPV-induced inhibition of p53. Although 14 cases possessed missense and deletion mutations of the p53 gene (of which four mutations were found in HPV-positive ESCC), no accumulation of the mutation was defined in the phenotype, suggesting that distinct mutation processes might be involved in HPV-negative and -positive ESCC. The data provide significant support for the hypothesis that HPV infection may play a crucial role in the oncogenesis of some ESCC.

Ploidy in human papillomavirus positive and negative vulvar squamous cell carcinomas and adjacent skin lesions.

To better characterize the two clinicopathologic types of squamous cell carcinoma, human papillomavirus (HPV) positive and negative, and their adjacent skin changes, we performed cytomorphometric analysis on 12 HPV-positive squamous cell carcinomas and adjacent vulvar intraepithelial neoplasia and 22 HPV-negative squamous cell carcinomas and adjacent squamous cell hyperplasia and lichen sclerosis. Our results were that 83% (10 of 12) HPV-positive carcinomas and 78% (7 of 9) adjacent vulvar intraepithelial neoplasia were aneuploid, compared with 59% (13 of 22) HPV-negative carcinomas, 6% (1 of 16) squamous cell hyperplasias and 0% (0 of 20) lichen scleroses. Seventy-five percent (9 of 12) HPV carcinomas and 78% (7 of 9) vulvar intraepithelial neoplasias showed two aneuploid peaks, but no HPV-negative carcinoma or non-neoplastic epithelial lesion showed multiple aneuploid peaks. Fifty percent of squamous cell hyperplasias (8 of 16) and lichen scleroses (10 of 20) adjacent to HPV-negative carcinomas were hypodiploid. The mean DNA indices were: 1.70 for the dominant tumor cell population of HPV-positive carcinoma, 1.64 for the dominant population of vulvar intraepithelial intraepithelial neoplasia, 1.41 for HPV-negative carcinoma, 0.85 for squamous cell hyperplasia and 0.83 for lichen sclerosis. In conclusion, the higher rate of aneuploidy, higher mean DNA index, and presence of multiploid peaks in HPV-positive carcinomas and adjacent vulvar intraepithelial neoplasias compared with the lower rate of aneuploidy, lower mean DNA index, absence of multiploid peaks of HPV-negative carcinomas and tendency to hypodiploidy in squamous cell hyperplasia and lichen sclerosis support the hypothesis that there are two clinicopathologic types of vulvar carcinoma, with different pathogenetic mechanisms. The similarities in ploidy findings between vulvar HPV-positive carcinomas and vulvar intraepithelial neoplasia and those previously published for cervical carcinoma and cervical intraepithelial neoplasia support the view that these two cancers are analogous and have similar pathogenetic mechanisms. The frequent finding of hypodiploidy in squamous cell hyperplasia and lichen sclerosis next to HPV-negative carcinomas requires further investigation of the molecular pathogenesis of this cancer type.

Prevalence of mucosotropic human papillomaviruses in squamous-cell carcinoma of the head and neck.

The prevalence of mucosotropic human papillomavirus (HPV) DNA in 63 squamous-cell carcinomas (SCC) from different anatomic sites in the head and neck was determined by general primer-mediated polymerase chain reaction (GP-PCR). HPV DNA was detected in 20.6% of SCC. Additional type-specific PCR for HPV 6, 11, 16, 18, 31 and 33 demonstrated the presence of HPV 16 alone in these carcinomas. HPV 16 was also detected in normal epithelium from the resection margins of the majority of HPV-positive SCC. HPV status did not correlate with tumour site, whether primary or recurrent, TNM stage, metastases, degree of differentiation, smoking or alcohol history, fate or survival.

Frequent mutations of p53 gene in oesophageal squamous cell carcinomas with and without human papillomavirus (HPV) involvement suggest the dominant role of environmental carcinogens in oesophageal carcinogenesis.

Epidemiological evidence suggests that alcohol intake, use of tobacco, ingestion of mycotoxins and nitrosamines and nutritional deficiencies are high-risk factors for the development of oesophageal cancer. Similarly, viral infections have been postulated to play a role in some tumours. However, the molecular events underlying the development of oesophageal carcinoma are poorly understood as yet. Loss of p53 tumour-suppressor gene function has been found in different human malignancies, and it can occur in a variety of ways, including gene mutation and interaction with the E6 protein of oncogenic human papillomaviruses (HPVs). Because the oesophageal mucosa is potentially exposed to mutagens and HPVs, we studied DNA samples derived from nine HPV-positive squamous cell carcinomas and 12 HPV-negative tumours. Exons 5-9 of the p53 gene containing phylogenetically conserved domains were examined using the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique. HPV detection was done using DNA in situ hybridisation with biotin-labelled HPV DNA probes. Mutations were detected in eight (38%) out of the 21 cases. Three mutations were found in exons 5/6, three in exon 7 and two in exon 8/9. Six (50%) of the 12 HPV-negative carcinomas showed p53 mutations. Two (22.2%) of the nine HPV-positive carcinomas were found to contain p53 mutations as well; one contained HPV 16 DNA sequences and showed p53 mutation in exon 8/9, and the other was HPV 6/11 positive with the mutation in exon 5/6. Although mutations were more common in HPV-negative tumours (50.0% vs 22.2%), the difference in p53 mutations in HPV-positive and -negative tumours did not reach statistical significance (P = 0.1946). These data indicate that inactivation of the p53 gene is a frequent event in oesophageal squamous cell carcinomas and such an inactivation might be an important molecular pathway for the development of oesophageal cancer. The findings of p53 mutations in HPV-positive oesophageal carcinomas suggest that HPV and p53 mutation were not mutually exclusive events. The presence of frequent mutations of p53 gene in both HPV-positive and -negative oesophageal carcinomas suggests a dominant role of environmental carcinogens in oesophageal carcinogenesis.