Kenji Nakamori

Multivariate analysis of occult lymph node metastasis as a prognostic indicator for patients with squamous cell carcinoma of the oral cavity

The biologic aggressiveness of squamous cell carcinoma of the oral cavity is reflected in its ability to metastasize to regional cervical lymph nodes. Patients with clinically negative cervical lymph nodes are believed to have a good prognosis; however, the prognosis of patients with lymph node metastasis occurring after excision or radiotherapy of the primary tumor is poor.

Clinical Assessment of the Relationship Between the Third Molar and the Inferior Alveolar Canal Using Panoramic Images and Computed Tomography

PURPOSE To predict the relationship between lower third molars and the inferior alveolar canal (IAC) from panoramic radiographs, and to establish criteria for using computed tomography (CT). MATERIALS AND METHODS A retrospective cohort study was performed involving 443 patients (695 teeth). Predictor variables were the distance between the third molar and the IAC, and findings according to the Roods criteria. Outcome variables were the absence of cortication between the third molar and the IAC on the CT image, and injury of the inferior alveolar nerve (IAN). Statistical analysis was performed to assess the relationship between predictor and outcome variables. RESULTS All patients had preoperative panoramic radiographs, and 71 patients (119 teeth) also had CT images. On CT examination, 48 teeth (40.3%) showed absence of cortication. Injury of the IAN was reported in 7 cases (1.0%), 5 of which exhibited absence of cortication; the remaining 2 did not have CT scans. Five of the 48 cases showing absence of cortication exhibited IAN injury, and none of the cases with cortication exhibited IAN injury. On the panoramic images, the following signs were strongly correlated with absence of cortication: a superimposed relationship between the third molar and the IAC; darkness of the root; and diversion and narrowing of the IAC. CONCLUSION Presence of Roods criteria was a predictor for a contact relationship between the third molar and the IAC, and an indication for CT examination. However, a superimposed relationship and the absence of Roods criteria did not necessarily signify a separate relationship between third molar and the IAC.

Phase I clinical trial of survivin‐derived peptide vaccine therapy for patients with advanced or recurrent oral cancer

Survivin, a member of the inhibitor of apoptosis protein (IAP) family, is abundantly expressed in most malignancies, but is hardly detectable in normal adult tissues. Previously we have identified a human leukocyte antigen (HLA)‐A24‐restricted antigenic peptide, survivin‐2B80‐88 (AYACNTSTL), recognized by CD8+ cytotoxic T lymphocytes (CTL). Survivin‐2B80‐88‐specific CTL were induced efficiently from peripheral blood mononuclear cells (PBMC) of oral cancer patients after stimulation with the peptide in vitro. We conducted a phase I clinical study to evaluate the safety and the efficacy of survivin‐2B80‐88 peptide vaccination in HLA‐A24‐positive patients with advanced or recurrent oral cancer. The vaccines were given subcutaneously or intratumorally six times at 14‐day intervals. Eleven patients were enrolled and 10 patients completed the vaccination protocol. No adverse events were observed in any patients. In two patients, the levels of serum squamous cell carcinoma (SCC) antigen decreased transiently during the period of vaccination. Tumor regression that was compatible with a partial response (PR) was noted in one patient. The remaining nine patients experienced progressive disease (PD). Immunologically, an increase of the peptide‐specific CTL frequency was detected in six of the eight patients evaluated by HLA‐A24/peptide tetramer analysis. The present clinical trial revealed that survivin‐2B peptide vaccination was safe and had therapeutic potential for oral cancer patients. However, subsequent clinical trials in combination with various adjuvant drugs will be required to improve the immunological and therapeutic efficacy. This trial was registered with University Hospital Medical Information Network (UMIN) number UMIN000000976. (Cancer Sci 2011; 102: 324–329)

High expression of ALDH1 and SOX2 diffuse staining pattern of oral squamous cell carcinomas correlates to lymph node metastasis.

One of the major factors involved in the prognosis of oral squamous cell carcinoma (OSCC) patients is metastasis. Recent progress in cancer stem‐like cell/cancer‐initiating cell (CSC/CIC) research indicates that CSCs are related to metastasis. Aldehyde dehydrogenase 1 – (ALDH1) and SRY‐related HMG‐box gene 2 (SOX2) have recently been shown to be putative CSC markers for several human malignancies. The aim of this study was to determine the association of ALDH1 and SOX2 expression in oral squamous cell carcinoma (OSCC) with lymph node metastasis. Immunohistochemical staining of ALDH1, SOX2 and Ki67 was performed in 80 OSCC tissues. High expression rates of ALDH1 (2%–40%) were found to be related to lymph node metastasis (P = 0.0017). Interestingly, we found that SOX2 staining could be classified into two patterns: (i) peripheral staining pattern; and (ii) diffuse staining pattern. The diffuse staining pattern showed a significant correlation with lymph node metastasis (P < 0.001). No correlation was found between Ki67 staining and lymph node metastasis (P = 0.4724). The ALDH1 positive staining rates in metastatic lymph nodes were higher than that in corresponding primary OSCC tissues. These results indicate that high expression rates of ALDH1 and SOX2 diffuse staining patterns might be novel prediction markers for OSCC lymph node metastasis.

Clinical significance of computed tomographic assessment and anatomic features of the inferior alveolar canal as risk factors for injury of the inferior alveolar nerve at third molar surgery.

PURPOSE To assess the clinical features of the inferior alveolar canal (IAC) using computed tomography (CT) and to analyze the significance of CT examination at third molar surgery. MATERIALS AND METHODS A retrospective cohort study was performed involving 99 patients (145 teeth). The relationship between cortication status, buccolingual position, and shape of the IAC on the CT image and inferior alveolar nerve (IAN) injury after third molar surgery were statistically analyzed. RESULTS The shape of the IAC was categorized into 3 groups: round/oval, teardrop, and dumbbell. IAN injury was observed in 7 of 145 cases (4.8%). All 7 cases exhibited absence of cortication; 3 were dumbbell shape and 4 were round/oval. According to logistic regression analysis of cases with absence of cortication, IAC shape was closely related to IAN injury. CONCLUSIONS These results suggest that assessment of the IAC shape and cortication status at third molar surgery may be clinically useful.

Carcinoma ex pleomorphic adenoma of the buccal region is composed of salivary duct carcinoma and squamous cell carcinoma components

A 73-year-old female presented with an asymptomatic mass in the left buccal region that she had first noticed 4 years earlier. The tumor, which was located in the buccal space, was clinically diagnosed as a salivary gland tumor and treated by excision. Histopathological examination revealed a capsule of connective tissue consisting of three different histopathological neoplastic areas in a large, fibrous, hyalinizing stromal background. The neoplastic lesion contained two malignant and one benign element, with histological characteristics consistent with squamous cell carcinoma (SCC), salivary duct carcinoma (SDC) and pleomorphic adenoma (PA). The SCC nests showed no continuity with the buccal mucosa. Both the SCC and SDC nests were surrounded by non-atypical myoepithelial cells, suggesting that both components may have developed from transformation of metaplastic luminal epithelial cells of PA. The tumor was diagnosed as a non-invasive carcinoma (SCC and SDC) ex pleomorphic adenoma (Ca-ex-PA). There was no evidence of recurrence 16 months after operation.

Assessment of the Shape of the Inferior Alveolar Canal as a Marker for Increased Risk of Injury to the Inferior Alveolar Nerve at Third Molar Surgery: A Prospective Study

PURPOSE Morphologic evaluation of computed tomographic images is an important assessment tool before surgical removal of the lower third molar (LM3). The aim of this study was to ascertain whether the shape of the inferior alveolar canal (IAC) is a reliable predictor for inferior alveolar nerve (IAN) injury during M3 surgery. MATERIALS AND METHODS This prospective study assessed samples with a high risk of IAN injury during M3 surgery based on orthopantomographic examination. The predictor variables were demographic factors (patients age and gender), anatomic factors (angulation of the tooth), and radiographic factors (cortication status, buccolingual position, shape of the IAC, number of roots, and root shape). The outcome variable was IAN injury. The relation between predictor and outcome variables was analyzed using the Fisher exact test and a logistic regression model. RESULTS One hundred sixty-nine LM3s (115 patients) were analyzed. IAN injury was observed in 12 of 115 patients and 13 of 169 LM3s (7.7%). All 13 cases with IAN injury exhibited absence of cortication. A dumb-bell-shaped IAC was considered a useful predictor for IAN injury (sensitivity, 69.2%; specificity, 84.6%). In cases with absence of cortication, logistic regression analysis indicated that a dumb-bell-shaped IAC was closely related to IAN injury (P = .005). CONCLUSION The cortication status and shape of the IAC are reliable predictors for IAN injury at M3 surgery. Cases exhibiting absence of cortication and a dumb-bell-shaped IAC should be recognized as presenting a high risk of IAN injury at M3 surgery.

Squamous cell carcinoma of the buccal mucosa in a young adult with history of allogeneic bone marrow transplantation for childhood acute leukemia

Secondary cancers are severe complications in patients who have had allogeneic bone marrow transplantation for childhood leukemia. We describe here a case of squamous cell carcinoma (SCC) of the buccal mucosa in a young adult patient who had had allogeneic bone marrow transplantation for childhood acute leukemia.

Small proline-rich protein-1B is overexpressed in human oral squamous cell cancer stem-like cells and is related to their growth through activation of MAP kinase signal.

Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are considered to be essential for tumor maintenance, recurrence and metastasis. Therefore, eradication of CSCs/CICs is essential to cure cancers. However, the molecular mechanisms of CSCs/CICs are still elusive. In this study, we investigated the molecular mechanism of the cell growth of oral CSCs/CICs. Oral CSCs/CICs were isolated as aldehyde dehydrogenase 1 bright (ALDH1(br)) cells by the ALDEFLUOR assay. Small proline-rich protein-1B (SPRR1B) gene was shown to be overexpressed in ALDH1(br) cells by a cDNA microarray and RT-PCR. SPRR1B was shown to have a role in cell growth and maintenance of ALDH1(br) cells by SPRR1B overexpression and knockdown experiments. To elucidate the molecular mechanism by which SPRR1B regulates cell growth, further cDNA microarray analysis was performed using SPRR1B-overexpressed cells and cells with SPRR1B knocked down by siRNA. Expression of the tumor suppressor gene Ras association domain family member 4 (RASSF4) was found to be suppressed in SPRR1B-overexpressed cells. On the other hand, the expression of RASSF4 was enhanced in cells in which SPRR1B expression was knocked down by SPRR1B-specific siRNA. RASSF4 has an RA (Ras association) domain, and we thus hypothesized that RASSF4 modulates the MAP kinase signal downstream of the Ras signal. MAP kinase signal was activated in SPRR1B-overexpressed cells, whereas the signal was suppressed in SPRR1B knocked down cells. Taken together, the results indicate that the expression of SPRR1B is upregulated in oral CSCs/CICs and that SPRR1B has a role in cell growth by suppression of RASSF4.

Hypoxia-induced epithelial-mesenchymal transition is regulated by phosphorylation of GSK3-β via PI3 K/Akt signaling in oral squamous cell carcinoma

OBJECTIVE Epithelial-mesenchymal transition (EMT) plays an important role in cancer invasion and metastasis induced by hypoxia. Here, we examined whether phosphorylation of GSK3-β via phosphoinositide 3-kinase (PI3 K)/Akt signaling is involved in enhancing the hypoxia-induced EMT in oral squamous cell carcinoma (OSCC). STUDY DESIGN Experiments were performed in OSCC cell lines (HSC-2, HSC-3, HSC-4, SAS, and HO-1-U-1) under normoxic or hypoxic conditions. The EMT was assessed by Matrigel invasion assays and wound healing assays. OSCC cell lines (HSC-2 and HSC-4) overexpressing hypoxia-inducible factor (HIF)-1α were established to examine the effects of HIF-1α on EMT-related factors. Immunohistochemical staining was performed to examine phosphorylation of GSK3-β in 33 cases of tongue squamous cell carcinoma. RESULTS Under hypoxic conditions, OSCC cell lines exhibited HIF-1α expression and showed evidence of the EMT. In cells overexpressing HIF-1α, the levels of phospho-Akt and phospho-GSK3-β were increased, resulting in induction of the EMT. Inhibition of GSK3-β phosphorylation suppressed these effects. Moreover, the intensity of pGSK3-β staining was significantly increased with cN stage and cTNM stage in patients with tongue squamous cell carcinoma. CONCLUSIONS Our data showed that the hypoxia-induced EMT in OSCC was enhanced by GSK3-β phosphorylation, suggesting that GSK3-β may be important in the invasion and metastasis of OSCC.