Naoki Otsuki

Depressed expression of Klotho and FGF receptor 1 in hyperplastic parathyroid glands from uremic patients.

Fibroblast growth factor 23 (FGF23) exerts its effect by binding to its cognate FGF receptor 1 (FGFR1) in the presence of its co-receptor Klotho. Parathyroid glands express both FGFR1 and Klotho, and FGF23 decreases parathyroid hormone gene expression and hormone secretion directly. In uremic patients with secondary hyperparathyroidism (SHPT), however, parathyroid hormone secretion remains elevated despite extremely high FGF23 levels. To determine the mechanism of this resistance, we measured the expression of Klotho, FGFR1, and the proliferative marker Ki67 in 7 normal and 80 hyperplastic parathyroid glands from uremic patients by immunohistochemistry. All uremic patients had severe SHPT along with markedly high FGF23 levels. Quantitative real-time reverse transcription PCR showed that the mRNA levels for Klotho and FGFR1correlated significantly with their semi-quantitative immunohistochemical intensity. Compared with normal tissue, the immunohistochemical expression of Klotho and FGFR1 decreased, but Ki67 expression increased significantly in hyperplastic parathyroid glands, particularly in glands with nodular hyperplasia. These results suggest that the depressed expression of the Klotho-FGFR1 complex in hyperplastic glands underlies the pathogenesis of SHPT and its resistance to extremely high FGF23 levels in uremic patients.

Value of retrospective image fusion of 18F-FDG PET and MRI for preoperative staging of head and neck cancer: Comparison with PET/CT and contrast-enhanced neck MRI

PURPOSE To assess the clinical value of retrospective image fusion of neck MRI and (18)F-fluorodeoxyglucose ((18)F-FDG) PET for locoregional extension and nodal staging of neck cancer. MATERIALS AND METHODS Thirty patients with carcinoma of the oral cavity or hypopharynx underwent PET/CT and contrast-enhanced neck MRI for initial staging before surgery including primary tumor resection and neck dissection. Diagnostic performance of PET/CT, MRI, and retrospective image fusion of PET and MRI (fused PET/MRI) for assessment of the extent of the primary tumor (T stage) and metastasis to regional lymph nodes (N stage) was evaluated. RESULTS Accuracy for T status was 87% for fused PET/MRI and 90% for MRI, thus proving significantly superior to PET/CT, which had an accuracy of 67% (p=0.041 and p=0.023, respectively). Accuracy for N status was 77% for both fused PET/MRI and PET/CT, being superior to MRI, which had an accuracy of 63%, although the difference was not significant (p=0.13). On a per-level basis, the sensitivity, specificity and accuracy for detection of nodal metastasis were 77%, 96% and 93% for both fused PET/MRI and PET/CT, compared with 49%, 99% and 91% for MRI, respectively. The differences for sensitivity (p=0.0026) and accuracy (p=0.041) were significant. CONCLUSION Fused PET/MRI combining the individual advantages of MRI and PET is a valuable technique for assessment of staging neck cancer.

Prevalence of Human Papillomavirus in Oropharyngeal Cancer: A Multicenter Study in Japan

Background: The incidence rates of oropharyngeal squamous cell carcinoma (OPSCC) have risen steadily in the USA and in northern Europe. These increases are thought to be a consequence of persistent infection with high-risk human papillomavirus (HPV) in OPSCC patients. HPV is an emerging etiologic factor in OPSCC. In Japan, the incidence of OPSCC has significantly increased over the last three decades. However, the population of HPV-positive OPSCC patients is currently unknown. We examined the nationwide trends with regard to HPV incidence in OPSCC patients at 21 specific sites, and examined the relationship between the presence of HPV and survival in OPSCC patients in Japan. Methods: Tumor samples were obtained from patients with OPSCC prior to treatment, and HPV infection was investigated by polymerase chain reaction (PCR). Hybrid Capture 2 (HC2) was also adopted for swab examination on the surface of fresh tumors. Results: HPV was detected by PCR in 79 (50.3%) out of 157 OPSCC patients. The clinical features of HPV-positive OPSCC were low differentiation, a tendency to involve the lateral wall, and high nodal staging. The sensitivity and specificity of HC2 were 93.7 and 96.2%, respectively, indicating its utility as a screening test. HPV-positive patients had significantly better overall survival and disease-free survival than HPV-negative patients.

Retropharyngeal node metastasis from papillary thyroid carcinoma

Papillary thyroid carcinomas commonly metastasize to paratracheal and jugular lymph nodes. Metastasis to the retropharyngeal node is rare for this tumor.

Voice quality after surgical treatment for thyroid cancer.

BACKGROUND Thyroidectomy is a standard treatment for thyroid cancers. Hoarseness due to the paralysis of the recurrent laryngeal nerve is one of the most common postoperative complications, and has been studied by many investigators. However, voice quality after thyroidectomy in patients in whom recurrent laryngeal nerves were preserved and vocal cord morbidity was endoscopically normal has not been well studied. To understand voice quality after thyroidectomy further, we conducted a time-course analysis of voice quality in patients who had thyroidectomy with normal cord morbidity by various measures. METHODS We evaluated voice parameters including the Voice Handicap Index-10 (VHI-10), the vocal efficacy index, the fundamental frequency (F0), the maximum phonation time (MPT), the mean air flow rate (MFR), jitter, shimmer, and the noise-to-harmonics ratio (NHR) before and after total thyroidectomy (TT) or lobectomy (LO) for thyroid cancers in 110 patients in whom the recurrent laryngeal nerves were preserved without apparent injury and normal vocal cord mobility was confirmed by endoscopic examination. Thirteen patients who underwent parotidectomy were enrolled as controls. RESULTS Immediately after surgery, significant decreases in MPT (p=0.003) and significant increases in jitters, shimmers, and NHR (p=0.0002, 0.02, and 0.03, respectively) were observed in the patients who underwent TT. In comparison with the controls, jitters and NHR were significantly higher in the patients who had a TT (p=0.03, 0.04). MFR was significantly higher in the patients who had an LO than in the controls (p=0.02). As compared with the patients who had an LO, MPT was significantly shorter (p=0.0004) and MFR and NHR were significantly higher (p=0.004, 0.03) in the patients with a TT. In the patients who had a TT, the MPT immediately after the surgery was significantly longer in the patients who had simultaneously neck dissection (ND) in comparison with the patients who did not have ND. However, all these differences gradually decreased and were not significant at one month after surgery. CONCLUSIONS Our results suggest that TT and ND have a distinct impact on voice quality after surgical treatment for thyroid cancer, probably due to slight and transient nerve conduction disorders induced by the manipulation around recurrent laryngeal nerves and/or laryngeal edema induced by the disturbance of venous and lymphatic drainages. However, these changes appear to be temporary, lasting only a few weeks.

Nuclear factor-κB expression as a novel marker of radioresistance in early-stage laryngeal cancer†‡

The aim of this study was to evaluate the significance of nuclear factor‐kappa B (NF‐κB) expression as a marker of radioresistance in early‐stage laryngeal cancer.

Particle Radiotherapy Using Protons or Carbon Ions for Unresectable Locally Advanced Head and Neck Cancers with Skull Base Invasion

OBJECTIVE To study the oncological outcome of the patients with unresectable locally advanced primary head and neck cancers invading the skull base, treated with particle radiotherapy. METHODS Fifty-seven patients with unresectable primary head and neck cancers invading the skull base received proton or carbon ion radiotherapy as definitive treatment at Hyogo Ion Beam Medical Center between 2003 and 2009. Forty-seven patients were treated with proton radiotherapy and 10 patients were treated with carbon ion radiotherapy. A retrospective review was performed with clinical charts and recorded imagings. RESULTS With a median follow-up of 32 months, the 3-year actual survival and local progression-free rates of all the patients were 61 and 56%, respectively. The 3-year actual survival rates of adenoid cystic carcinoma, squamous cell carcinoma, olfactory neuroblastoma, adenocarcinoma and malignant melanoma were 83, 44, 75, 0 and 38%, respectively. The 3-year actual local control rates of adenoid cystic carcinoma, squamous cell carcinoma, olfactory neuroblastoma, adenocarcinoma and malignant melanoma were 63, 31, 83, 50 and 0%, respectively. Distant metastasis was observed in 13 of 25 patients in adenoid cystic carcinoma, two of 14 patients in squamous cell carcinoma, one of six patients with olfactory neuroblastoma, two of four patients with adenocarcinoma, three of four patients with malignant melanoma and two of three patients with undifferentiated carcinoma. Mucositis and dermatitis were seen as acute toxicities. The most common late toxicity was visual disorder. Grades 2, 3 and 4 visual disorders were observed in seven, five and two patients, respectively. CONCLUSIONS Proton and carbon ion radiotherapy resulted in satisfactory local control in patients with locally advanced unresectable primary head and neck cancers invading the skull base.

Gene silencing with siRNA targeting E6/E7 as a therapeutic intervention against head and neck cancer-containing HPV16 cell lines

Abstract Conclusion: Our results indicate that siRNA E6 and/or E7 may have potential as a gene-specific therapy for human papillomavirus (HPV) type 16 (HPV16)-related squamous cell carcinoma of the head and neck (HNSCC). Objectives: To evaluate the effectiveness of siRNA targeting E6 and/or E7 on the in vitro and in vivo growth suppression of HPV16-related HNSCC. Methods: HPV16-related HNSCC (UM-SCC47) cell lines were used for the present study. Expression of HPV viral oncogenes E6 and/or E7 and their cellular targets, p53 and pRb, was evaluated by real-time PCR, Western blotting, and immunofluorescence staining. To study the effect of siRNA on tumor growth in vivo, we developed animal models. Representative tumors harvested from each group were processed for apoptosis analyses (TUNEL assay) and immunofluorescence staining for p53 and pRb. Results: E6 and E7 oncogenes of HPV16 were down-regulated by E6 and/or E7 targeting siRNAs, respectively. The expression of p53 and pRb proteins in both the E6 siRNA group and E7 siRNA group was up-regulated compared with those of control groups. The cellular proliferation and apoptosis indexes of E6 and/or E7 siRNA groups were higher than those of controls. In vivo studies showed significant inhibitory effect of E6 and/or E7 siRNA compared with those of control groups, which was consistent with in vitro studies.

Prognostic Value of FDG PET Imaging in Patients with Laryngeal Cancer

Background and Purpose To investigate the prognostic value of 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) in patients with laryngeal cancer. Materials and Methods The study included 51 patients of whom 30 underwent definitive radiotherapy with or without chemotherapy and 21 underwent radical surgery with or without adjuvant chemoradiation therapy. FDG uptake by both the primary lesion and the neck node was measured using the maximum standardized uptake value (SUVmax). The effects of clinicopathological factors including primary tumor SUVmax and nodal SUVmax on progression-free survival, local control, nodal progression-free survival, and distant metastasis-free survival were evaluated using the log-rank test and Cox method. Results The median duration of follow-up was 48.6 months (range 8 to 82.1 months). Univariate analysis showed that nodal SUVmax, N status, and tumor TNM stage were significantly associated with recurrence, whereas primary tumor SUVmax, age, treatment strategy and T status were not. Multivariate analysis demonstrated that only the nodal SUVmax was a significantly unfavorable factor for progression-free survival (p = 0.029, hazard ratio 0.54, 95% CI 0.38-0.87) and nodal progression-free survival (p = 0.023, hazard ratio 0.51, 95% CI 0.34-0.81). ROC curve analysis and log-rank test showed that patients with a high nodal SUVmax (≧4) had a significantly lower progression-free survival rate than those with a low SUVmax (<4; p<0.0001). Conclusions The pretreatment SUVmax of nodal disease in patients with laryngeal cancer is prognostic for recurrence.

E10A, an adenovirus-carrying endostatin gene, dramatically increased the tumor drug concentration of metronomic chemotherapy with low-dose cisplatin in a xenograft mouse model for head and neck squamous-cell carcinoma.

Most cancer chemotherapeutic agents are administered at the maximum-tolerated dose (MTD) in short cycles with treatment breaks. However, MTD-based chemotherapies are often associated with significant toxicity and treatment breaks allow the opportunity for tumor regrowth and acquisition of chemoresistance. To minimize these drawbacks, a metronomic strategy, in which chemotherapeutics are administered at doses significantly below the MTD without treatment breaks, has been suggested by many investigators. The antitumor effect of metronomic chemotherapy may be partially due to inhibition of tumor angiogenesis, and it could be enhanced by a combination therapy, including antiangiogenic agents. In this study, we evaluated the synergistic effect of E10A, an adenovirus carrying the endostatin gene, the most potent inhibitors of tumor angiogenesis, in combination with weekly low-dose cisplatin in a xenograft mouse model for head and neck squamous-cell carcinoma. The E10A induced mRNA and protein expressions of endostatin in H891 cells in vitro. E10A significantly enhanced the in vivo tumor growth inhibitory effect of cisplatin. Immunohistochemical analysis with a TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) assay and anti-CD31 antibodies revealed that the combination of E10A and cisplatin induced high levels of cell apoptosis and inhibited tumor angiogenesis. Importantly, E10A increased the platinum concentrations in tumors to fivefold higher than that induced by cisplatin alone.