Zeyi Deng

Viral load, physical status, and E6/E7 mRNA expression of human papillomavirus in head and neck squamous cell carcinoma

The purpose of this study was to determine prospectively both human papillomavirus (HPV) load and physical status in different types of head and neck squamous cell carcinoma (HNSCC).

Prognostic value of human papillomavirus and squamous cell carcinoma antigen in head and neck squamous cell carcinoma

To clarify the synergistic influence of human papillomavirus (HPV) status and squamous cell carcinoma antigen (SCCA) mRNA expression on head and neck squamous cell carcinoma (HNSCC) prognosis, HPV DNA presence and SCCA1 and SCCA2 mRNA expression were determined by PCR and quantitative real‐time RT‐PCR, respectively, in 121 patients with primary HNSCC who were receiving curative treatment. HPV DNA was detected in 28.1% (34/121) of HNSCC cases, and only high‐risk types (HPV‐16, HPV‐33, HPV‐35 and HPV‐58) were observed. Positive HPV status showed a significantly better prognosis than negative HPV status (P = 0.022). An elevated SCCA2/SCCA1 mRNA ratio was an independent predictor of disease recurrence (P = 0.004). In addition, HPV‐negative patients with a high SCCA2/SCCA1 ratio (>0.27) had a significantly lower recurrence‐free survival rate than HPV‐negative patients with a low SCCA2/SCCA1 ratio (P < 0.011). Our findings revealed that both HPV status and the SCCA2/SCCA1 mRNA ratio are independently associated with prognosis in HNSCC. Patients with both a HPV‐negative status and a high SCCA2/SCCA1 ratio might need intensified treatment and rigorous follow up after treatment because of the high risk of recurrence.

Oral candidiasis in patients receiving radiation therapy for head and neck cancer

OBJECTIVE: To investigate oral candidiasis in patients with head and neck cancer before, during, and after radiation therapy, and to explore its association with clinical oropharyngeal symptoms. STUDY DESIGN: A cohort study. SETTING: University hospital. SUBJECTS AND METHODS: Subjects who received radiation therapy (RT) for the treatment of head and neck cancer were divided into two groups: an oral cavity irradiated group (OIRR group, n = 29) and an oral cavity nonirradiated group (ONIRR group, n = 17). A control group consisted of 18 healthy subjects. Patients were examined for signs of oral candidiasis before, during, immediately after, and one month after RT. Mouth and throat soreness (MTS), dysphagia, and xerostomia were evaluated by self-reported questionnaires, and associations between oral candidiasis and these symptoms were analyzed. RESULTS: The incidence of oral candidiasis during RT was significantly higher in the OIRR group (55.2%) than in the ONIRR group (11.8%). Similarly, the occurrence of xerostomia during RT was significantly higher in the OIRR group (86.2%) than in the ONIRR group (52.9%). In the OIRR group, the mean MTS score at the 20th fraction of RT was significantly higher in patients with candidiasis (mean ± SD, 5.8 ± 2.1) than in those with RT-induced mucositis without candidiasis (3.7 ± 2.0). In the OIRR group, 65.2 percent of patients who experienced dysphagia developed oral candidiasis, compared with only 10 percent in the ONIRR group. CONCLUSION: Oral candidiasis concurrent with oral mucositis due to RT may increase oropharyngeal discomfort during RT.

Human papillomavirus load and physical status in sinonasal inverted papilloma and squamous cell carcinoma.

BACKGROUND This study investigated prospectively the role of human papillomavirus (HPV) in paranasal inverted papilloma (IP). METHODS HPV presence and viral load and physical status of HPV-16 were examined by polymerase chain reaction-based methods using fresh frozen samples obtained from 13 patients with IP (IP group), 11 with squamous cell carcinoma in the maxillary sinus (SCC group) and 39 with chronic inflammatory lesions (inflammatory group). RESULTS The presence of the HPV genome was detected in 46.1%, 27.3% and 7.6% of patients in the IP, SCC and inflammatory groups, respectively. The IP group showed significantly higher HPV-positive rates than the inflammatory group. All types of HPV detected were high-risk HPV, especially HPV-16. The relative HPV-16 copy numbers varied from 2.5 to 1524.1 per 50 ng genomic DNA. The viral load was higher in the IP and SCC groups than in the inflammatory group. In the IP group, no significant relationship was found between HPV-16 viral load and clinical characteristics, or between physical status and clinical characteristics. One patient with IP and concomitant squamous cell carcinoma, however, showed high viral load and integration. CONCLUSIONS HPV infection is involved in the pathogenesis of IP, and high viral load and integration of HPV have an important role in malignant lesion in association with IP.

Squamous cell carcinoma antigen production in nasal inverted papilloma.

Background The clinical importance of serum squamous cell carcinoma antigen (SCCA) and SCCA subclasses has not been established for treating inverted papilloma (IP). The aim of this study was to clarify the clinical importance of serum SCCA and its subclasses in IP, compared with maxillary squamous cell carcinoma and inflammatory disease. Methods Serum SCCA was measured in 22 patients with IP (IP group), 11 with maxillary squamous cell carcinoma (carcinoma group), and 22 with inflammatory disease (inflammatory group). mRNA expression of SCCA subclasses was examined using quantitative real-time polymerase chain reaction. Results In the IP group, 81.8% showed elevated serum SCCA, and 90.3% with recurrent IP showed elevated SCCA. The preoperative SCCA value (mean ± SD, 3.99 ± 4.39) in the IP group was significantly higher than in the carcinoma (1.28 ± 0.88; p = 0.012) and inflammatory (0.60 ± 0.31; p < 0.001) groups. mRNA expression of SCCA1 and SCCA2 in the IP group was higher than in the carcinoma and inflammatory groups. The SCCA2/SCCA1 ratio of mRNA expression (0.11 ± 0.06) in the IP group was similar to that (0.11 ± 0.09) in the inflammatory group, although the ratio (0.20 ± 0.12) in the carcinoma group was significantly higher than in the IP and inflammatory groups The receiver operating characteristic curve analysis for the SCCA2/SCCA1 ratio to detect carcinoma yielded an area under the curve of 0.760 (95% confidence interval, 0.626–0.894). Conclusion The serum level of SCCA is effective for detecting IP, including recurrent IP. In contrast, the SCCA2/SCCA1 ratio is useful for detecting squamous cell carcinoma among other sinonasal diseases.

Prediction of concurrent chemoradiotherapy outcome in advanced oropharyngeal cancer

The aim of this study was to investigate human papillomavirus (HPV) infection as a predictor of concurrent chemoradiotherapy (CCRT) response and indicator of planned neck dissection (PND) for patients with advanced oropharyngeal squamous cell carcinoma (OPSCC; stage III/IV). Overall, 39 OPSCC patients (32 men, 7 women; median age 61 years, range 39–79 years) were enrolled. The primary lesion and whole neck were irradiated up to 50.4 Gy, and subsequently the primary site and metastatic lymph nodes were boosted with a further 16.2 Gy. Although several chemotherapy regimens were employed, 82.1% of OPSCC patients received the combination of nedaplatin and 5-fluorouracil. HPV-related OPSCC (16 cases) was defined as both HPV DNA-positive status by polymerase chain reaction and p16INK4a overexpression by immunohistochemistry. Patients with N2 and N3 disease received PND 2–3 months after CCRT completion. Compared to non-responders, CCRT responders showed significantly lower nodal stage (N0 to N2b) and HPV-positive status in univariate analysis. Patients with HPV-related OPSCC had longer time to treatment failure (TTF) than those with HPV-unrelated OPSCC (p=0.040). Three-year TTF was 81.3 and 47.8% in the HPV-related and HPV-unrelated groups, respectively. There were also significant differences in disease-free survival (DFS) between the two OPSCC patient groups (p=0.042). Three-year DFS was 93.8 and 66.7% in patients with HPV-related and HPV-unrelated OPSCC, respectively. Multivariate logistic analysis showed a lower risk of TTF event occurrence in HPV-related OPSCC (p=0.041) than in HPV-unrelated OPSCC. Thus, HPV testing in addition to nodal stage was useful for predicting CCRT response, especially in advanced OPSCC. Because patients who received PND showed moderate locoregional control, PND is an effective surgical procedure for controlling neck lesions in patients with advanced HPV-unrelated disease.

Squamous cell carcinoma antigen as a diagnostic marker of nasal inverted papilloma.

Background Serum squamous cell carcinoma antigen (SCCA) levels are elevated in sinonasal inverted papilloma (IP). However, the relationship between tumor volume and SCCA level, and the influence of skin or pulmonary diseases in which the SCCA level is high, have not been established. Objective To clarify whether the level of serum SCCA can be used as a diagnostic marker of IP. Methods Serum SCCA level was measured in 30 patients with IP (IP group) and 57 with inflammatory disease (inflammatory group). Results Overall, 83.3% in the IP group showed elevated serum SCCA levels regardless of whether they were new patients or patients with recurrent IP, and SCCA levels rapidly decreased after surgery. Only 5.3% had elevated SCCA levels in the inflammatory group. Before surgery, the IP group had a median preoperative SCCA level of 2.4 ng/mL, whereas the median preoperative SCCA level was 0.9 ng/mL in the inflammatory group. Pre- and postoperative SCCA levels were significantly different in the IP group. With regard to the IP diagnosis in the IP and inflammatory groups based on the SCCA level (ng/mL), sensitivity and specificity were 83.3% and 94.7%, respectively. There was no significant correlation between SCCA elevation and respiratory function, and skin disease in the two groups, except for smoking in the IP group. Preoperative SCCA levels were significantly higher in smokers than in never-smokers in the IP group. Tumor volume was significantly correlated with SCCA level in IP. Multivariable logistic analysis showed that tumor volume was a predictor of preoperative SCCA elevation (p = 0.036; 95% confidence interval, 1.027–2.176). Conclusion Serum SCCA level is a reliable diagnostic marker to distinguish new and recurrent IP from inflammatory disease. Because smokers tended to have higher SCCA levels in IP, a different cutoff level might be needed. Although respiratory dysfunction and skin disease were not related to SCCA level, they should be taken into consideration when evaluating SCCA level.

Detection of human papillomavirus in branchial cleft cysts

High-risk human papillomavirus (HPV) DNA has been reported to be present in branchial cleft cysts, but further information is required to clarify the role of HPV infection in branchial cleft cysts. The presence of HPV, the viral load and the physical statuses in samples from six patients with branchial cleft cysts were investigated using the polymerase chain reaction (PCR), quantitative PCR, in situ hybridization (ISH) using HPV DNA probes and p16INK4a immunohistochemical analysis. High-risk type HPV-16 DNA was identified in four of the six branchial cleft cysts analyzed. Of the HPV-positive branchial cleft cysts, three exhibited mixed-type integration of HPV. HPV DNA was distributed among the basal-to-granular layers of the cystic wall in ISH analysis, and p16INK4a was weakly expressed in the nuclei and cytoplasm of the same layers in patients with integration. ISH revealed that one patient with episomal-type infection exhibited HPV DNA in the cyst wall and did not express p16INK4a. Two patients without evidence of HPV infection exhibited weak p16INK4a expression in the superficial cyst-lining cells of branchial cleft cysts. These results indicate that infection with high-risk HPV types may be common in branchial cleft cysts. In addition, p16INK4a is not a reliable surrogate marker for HPV infection in branchial cleft cysts.

Methylation of CpG sites in the upstream regulatory region, physical status and mRNA expression of HPV-6 in adult-onset laryngeal papilloma

The methylation status of HPV-6 upstream regulatory region (URR) in adult-onset laryngeal papillomatosis (AO-LP) remains unclear. The purpose of this study was to investigate the methylation status of URR and the physical status of HPV-6, as well as the dynamic variations of viral load and mRNA expression in AO-LP. We examined 18 specimens from 11 patients with AO-LP by real-time polymerase chain reaction (PCR), bisulfite-sequencing PCR, and amplification of papilloma oncogene transcripts. HPV-6 was identified in 9 of 11 patients (81.8%), and all the 15 specimens derived from 9 HPV-6-positive cases contained only episomal HPV-6 transcripts with intact E2. Three HPV-6-positive patients developed recurrent lesions, and HPV-6 copy numbers and mRNA expression decreased after surgical treatment. Among the 96 CpG sites (16/case), 67 (69.8%) were unmethylated, while 23 (30.2%) were heterogeneous (≥ 1 methylated CpG clone). High viral loads and episomal status of HPV-6 were frequently observed in AO-LP; thus, persistent E6/E7 mRNA expression of LR-HPV-6 may be associated with AO-LP recurrences. Hypomethylation and scattered patterns of methylated CpGs at the URR of HPV-6 were identified in AO-LP.

Importance of Human Papillomavirus Integration in Malignant Transformation of Inverted Papilloma

Objectives: This study investigated the role of human papillomavirus (HPV) in malignant transformation of paranasal inverted papilloma (IP). Methods: HPV presence and viral load and physical status of HPV-16 were examined by polymerase chain reaction-based methods using fresh frozen samples or paraffin-embedded samples obtained from 17 patients with IP (IP group), 5 with IP and squamous cell carcinoma (IP+SCC group), 16 with SCC (SCC group), and 67 with chronic inflammatory lesions (inflammatory group). Results: The presence of the HPV genome was detected in 29.4%, 40.0%, 25.0%, and 6.0% of patients in the IP, IP+SCC, SCC, and inflammatory groups, respectively. The IP group showed significantly higher HPV-positive rates than the inflammatory group. All types of HPV detected were high-risk HPV, especially HPV-16. The relative HPV-16 copy numbers varied from 2.5 to 7953 per 50 ng genomic DNA. Viral load was higher in the IP+SCC group than in the inflammatory group. In the IP group, no significant relationship was found between HPV-16 viral load and clinical characteristics. All patients with IP+SCC and SCC showed integration of HPV-16. Conclusions: High viral load and integration of HPV have an important role in malignant lesion in association with IP.