Satoru Ozeki

Prevalence of Epstein–Barr virus in oral squamous cell carcinoma

Forty‐six samples of oral squamous cell carcinoma (OSCC) were evaluated for the prevalence of Epstein–Barr virus (EBV) infection by the polymerase chain reaction (PCR), Southern blot hybridization, and in situ hybridization (ISH). EBV DNA was detected in 7 (15·2 per cent) out of 46 samples by a combination of PCR and Southern blot hybridization methods. All seven positive samples showed well‐differentiated carcinoma, thus suggesting a possible relationship between EBV infection and the degree of differentiation of carcinoma tissue. Latent infection membrane protein 1 (LMP1) was detected immunohistochemically in six of the EBV‐positive OSCCs. However, no signal of the EBV‐encoded small RNA (EBER)‐1 was demonstrated by the ISH method. No significant relationship was observed between EBV infection and lymph node metastasis. A follow‐up study (range from 4·4 to 79 months; mean 34·9 months) showed no recurrence or death to occur in the EBV‐positive patients, which thus suggested a good prognosis for EBV‐positive OSCC patients. Copyright

Imaging findings of lipomas in the orofacial region with CT, US, and MRI

OBJECTIVE The aim of this study was to document retrospectively the imaging findings of lipomas with the use of computed tomography, ultrasonography, and magnetic resonance imaging. STUDY DESIGN Thirteen patients with 11 lipomas and 2 lipomatoses were evaluated. Eleven cases were examined by computed tomography, 9 by ultrasonography, and 3 by magnetic resonance imaging. RESULTS Lipomas had a density ranging from -134 to -83 Hounsfield units, (mean-108) on the computed tomography images. The margins were ill defined in 9 of 10 cases. The superficial muscles were displaced externally in 8 cases and internally in 2 cases. With ultrasonography, 8 lesions were hypoechoic, and one was hyperechoic. All three lesions had a high signal intensity on both T1- and T2-weighted images. CONCLUSIONS Lipomas had a specific range of computed tomography Hounsfield unit values and also displaced the surrounding soft tissue. Although some variation in the ultrasonographic appearance was observed, the lesions tended to be hypoechoic. These findings may be useful for diagnosing lipomas in the orofacial region.

Metastasis to the linguallymph node in carcinoma of the tongue

Lingual lymph nodes are occasionally found along the course of some lymph-vessels of the tongue. However, their existence has received little attention and metastasis of carcinoma of the tongue to them has not previously been reported. Three cases of carcinoma of the tongue with metastasis to the lingual lymph node are described and a discussion of the importance of such metastasis in the treatment of carcinoma of the tongue is presented.

Assessment of the Relationship Between Impacted Mandibular Third Molars and Inferior Alveolar Nerve With Dental 3-Dimensional Computed Tomography

PURPOSE The purpose of this study was to assess the capacity of dental 3-dimensional computed tomography (3D-CT; limited cone-beam CT) to predict the exposure and injury of the inferior alveolar nerve (IAN) after mandibular third molar extractions. MATERIALS AND METHODS This study was a retrospective case series of patients who presented for extraction of mandibular third molars. Subjects eligible for study enrollment were those who underwent preoperative dental 3D-CT because the mandibular third molars were determined to be extremely close to the IAN on panoramic radiogram. The predictive variable was the anatomic relation of the IAN and third molar apices and was a binary variable, contact or noncontact. The primary outcome variable was IAN exposure, and the secondary outcome variable was IAN injury. RESULTS From January 2006 to August 2007, 1,853 mandibular third molars in 1,539 patients were extracted. Among them, dental 3D-CT was performed on 53 third molars in 47 patients. The mandibular third molars were judged to make contact with the mandibular canal on dental 3D-CT images in 35 cases (66%). Intraoperative IAN exposure was observed in 17 (49%) contact cases and 2 (11%) noncontact cases on dental 3D-CT images. Of 53 cases extracted after dental 3D-CT examinations, IAN injury occurred in 8 cases (15%). IAN exposure led to IAN injury in 36.8% of cases, whereas IAN injury occurred in only 2.9% of cases without IAN exposure. Although the incidence of IAN injury in the molar-canal contact cases was 23%, all 8 cases with IAN injury (100%) were included in these contact cases. CONCLUSION When viewing the anatomic relation between the IAN and mandibular third molar root apices using dental 3D-CT, contact of the 2 anatomic structures results in an increased risk for IAN exposure or injury.

Oral squamous cell carcinoma cells induce osteoclast differentiation by suppression of osteoprotegerin expression in osteoblasts

The invasion of oral squamous cell carcinoma (SCC) cells into the mandibular bone is a common clinical problem. It has been reported that BHY cells, a human oral SCC cell line, are capable of invading mandibular bone of nude mice. These results led us to examine possible mechanisms of osteoclastogenesis induced by BHY cells using in vitro culture systems. When BHY cells were cocultured with mouse bone marrow cells (BMCs), only few osteoclasts were formed, even though BHY cells express the receptor activator of NF‐κB ligand (RANKL). However, adding BHY cells to a coculture of mouse primary osteoblasts (POBs) and BMCs markedly induced osteoclastogenesis in the absence of osteotropic factors. Furthermore, another oral SCC cell line, HSC‐2, which does not express RANKL, also induced osteoclastogenesis in our cocultures. These effects were significantly, but not completely, inhibited by adding osteoprotegerin (OPG). In addition, we also found that TNFα released from these cells partially contributes to osteoclastogenesis via a RANKL‐independent mechanism. Adding BHY or HSC‐2 cells suppressed mouse OPG mRNA expression and protein production by POBs in cocultures of POBs and human oral SCC cells. This finding is consistent with the result that BHY cells and HSC‐2 cells did not enhance osteoclastogenesis in cocultures of BMCs and POBs from OPG‐deficient mice. Immunohistochemical analysis showed a reduction of OPG expression in osteolytic lesions as compared to normal lesions from oral SCC patients. Therefore, oral SCC‐induced suppression of OPG expression in POBs appears critical for osteoclastogenesis, rather than expression of RANKL in SCC cells.

Cervical Lymph Node Metastasis in Carcinoma of the Tongue Correlation between Clinical and Histopathological Findings and Metastasis

The factors relating to lymph node metastasis occurring in association with carcinoma of the tongue have been investigated clinically and histopathologically by many authors. Nevertheless, accuracy in predicting lymph node metastasis has not yet been attained. The present study describes the results of analysis performed in an attempt to predict lymph node metastasis of carcinoma of the tongue. Ninety-eight patients with squamous cell carcinoma of the tongue who were treated at our institution were investigated. Among the 98 patients, lymph node metastases were histologically proven after neck dissection in 40 patients. For statistical analysis in the present study, the quantification theory Type II of Hayashi (1954) was employed for discriminatory analysis. According to this discriminatory analysis, the correlations between the clinical (age, tumour size, N-category) and histopathological (degree of keratinization, mitoses, structure, mode of invasion) findings and metastasis were assessed on the basis of the partial correlation coefficients; these coefficients for each finding were not greatly different from each other and 74 out of 98 patients (76%) could be differentiated between patients with metastasis and without metastasis accurately. The prediction of lymph node metastasis by such discriminatory analysis had significant validity.

Sonographic analysis of recurrent parotitis in children: a comparative study with sialographic findings.

OBJECTIVE The sonographic features of recurrent parotitis in children were studied to clarify a relationship between sonographic and sialographic findings in this disease. STUDY DESIGN Twenty-one glands (7 on follow-up) were examined by 7.5 MHz ultrasonography and sialography. Echo intensity level, distribution of the internal echoes, and size of hypoechoic areas were compared with the size of punctate shadows on the sialograms. Twenty other histopathologic specimens were analyzed to investigate the entity of hypoechoic areas. RESULTS Sonography showed hypoechoic, heterogeneous internal echoes, the level of which increased as the punctate shadows enlarged. Hypoechoic areas, all of which were larger than the sialographic punctate shadows, were observed in 62% of the glands. Histopathologic analysis suggests that these hypoechoic areas represent dilated peripheral ducts with lymphocytic infiltration. Sonography was likely to detect changes over time more sensitively than sialography. CONCLUSIONS Sonography should be performed as the test of first choice, both in the primary and follow-up stages, in cases of recurrent parotitis in children.

STAT3 signal transduction through interleukin-22 in oral squamous cell carcinoma

Interleukin (IL)-22 is a member of the IL-10 family. Its main targets are epithelial cells, not immune cells. We examined IL-22 signal transduction in oral squamous cell carcinoma (OSCC) cells. Immunohistochemical staining revealed that IL-22R was expressed more highly in OSCC compared to normal regions. An IL-22R signal was also observed in metastatic OSCC cells in the lymph node. RT-PCR showed that the human OSCC cell lines MISK81-5, HSC-3, HSC-4, SAS and SQUU-B expressed IL-22 receptor chains. Immunoblotting showed that IL-22 induced a transient tyrosine phosphorylation of STAT3 (pY705-STAT3) in MISK81-5 cells. The change in the serine phosphorylation of STAT3 was subtle during the examination periods. Simultaneously, pY705-STAT3 activation in HSC-3 cells was undetectable after IL-22 stimulation. Immunocytochemistry demonstrated that IL-22 induced the translocation of phosphorylated STAT3 into the nucleus of MISK81-5 cells. IL-22 temporarily upregulated the expression of anti-apoptotic and mitogenic genes such as Bcl-x, survivin and c-Myc, as well as SOCS3. IL-22 transiently activated ERK1/2 and induced a delayed phosphorylation of p38 MAP kinase, but negligibly involved the activation of NF-κB in MISK81-5 cells. MISK81-5 and SQUU-B cells treated with IL-22 showed mild cellular proliferation. MISK81-5, HSC-4 and SAS cells treated with IL-22 downregulated the keratinocyte differentiation-related genes compared with unstimulated cells. Conversely, STAT3 suppression by STAT3 siRNA strongly disrupted the down-regulation of these genes by IL-22, but it did not significantly affect the activation of ERK1/2 by IL-22. The OSCC cells used in this study upregulated the expression of SERPINB3/4 (SCCA1/2), well-known SCC markers, following treatment with IL-22. These results indicate that IL-22 differentially activates the STAT3 signaling system depending on the type of OSCC. IL-22 may therefore play a role in tumor growth, cell differentiation and progression through STAT3-dependent and -independent pathways.

Reactive oxygen species stimulates epithelial mesenchymal transition in normal human epidermal keratinocytes via TGF-beta secretion

Epithelial to mesenchymal transition (EMT) plays an important role in tumor progression, and is an early step in carcinogenesis. Although reactive oxygen species (ROS) are known to be implicated in EMT in many tumor cell types, its exact role in EMT initiation in normal human cells, especially epidermal keratinocytes (NHEKs), remains unknown. To clarify whether ROS induce EMT in NHEKs, and to establish how ROS regulate EMT, we examined the effect of hydrogen peroxide (H(2)O(2)) on the expression of molecules involved in EMT and cell morphology in NHEKs. H(2)O(2) altered the expression of EMT biomarkers, including downregulation of epithelial cadherin and upregulation of α-smooth muscle actin, through a transcriptional modulator, Snail1. H(2)O(2) also induced epithelial to fibroblast-like morphological changes, together with upregulation of EMT biomarkers, and promoted phosphorylation of ERK1/2 and JNK in a time-dependent manner. Interestingly, H(2)O(2) stimulated the expression and secretion of TGF-β1 in NHEKs. Exogenous TGF-β1 also induced the expression of EMT biomarkers. In contrast, neutralizing antibody anti-TGF-β1 antibody or inhibitor of TGF-β receptor type I suppressed the expression of EMT biomarkers. Our results suggest that ROS stimulated TGF-β1 secretion and MAPK activation, resulting in EMT initiation in NHEKs.

Effect of SCCA1 and SCCA2 on the Suppression of TNF-α-Induced Cell Death by Impeding the Release of Mitochondrial Cytochrome c in an Oral Squamous Cell Carcinoma Cell Line

This study examined the effect of squamous cell carcinoma antigen 1 (SCCA1) and SCCA2 on TNF-α-induced cell death in human oral squamous cell carcinoma cell lines. The viability of MISK81-5 and sMISK cells treated with TNF-α dose-dependently decreased. The sMISK cells which stably overexpressed SCCA1 and SCCA2 cDNA showed a greater resistance against the cell death induced by TNF-α than the controls. Immunocytochemical staining for cytochrome c showed a punctate pattern in the cytoplasm of the TNF-α-untreated cells. After treatment with TNF-α, the punctate staining pattern was preserved in the transfectants, while this pattern disappeared in the controls. In the transfectants, the release of cytochrome c from the mitochondria to cytosol after TNF-α treatment was lower than in the controls. A decreased level of active caspase-9 was also observed in the transfectants. These results suggest that both SCCA1 and SCCA2 play a role in the prevention of TNF-α-induced cell death in vitro, by inhibiting the release of mitochondrial cytochrome c to some degree.