Hirofumi Tomioka

Identification of a predictive gene expression signature of cervical lymph node metastasis in oral squamous cell carcinoma

An accurate assessment of the cervical lymph node metastasis status in oral cavity cancer not only helps predict the prognosis of patients, but also helps surgeons to perform the appropriate treatment. We investigated the utilization of microarray technology focusing on the differences in gene expression profiles between primary tumors of oral squamous cell carcinoma that had metastasized to cervical lymph nodes and those that had not metastasized in the hope of finding new biomarkers to serve for diagnosis and treatment of oral cavity cancer. To design this experiment, we prepared two groups: the learning case group with 30 patients and the test case group with 13 patients. All tissue samples were performed using laser captured microdissection to yield cancer cells, and RNA was isolated from purified cancer cells. To identify a predictive gene expression signature, the different gene expressions between the two groups with and without metastasis in the learning case (n = 30) were analyzed, and the 85 genes expressed differentially were selected. Subsequently, to construct a more accurate prediction model, we further selected the genes with a high power for prediction from the 85 genes using the AdaBoost algorithm. The eight candidate genes, DCTD, IL‐15, THBD, GSDML, SH3GL3, PTHLH, RP5‐1022P6 and C9orf46, were selected to achieve the minimum error rate. Quantitative reverse transcription–polymerase chain reaction was carried out to validate the selected genes. From these statistical methods, the prediction model was constructed including the eight genes and this model was evaluated by using the test case group. The results in 12 of 13 cases (∼92.3%) were predicted correctly. (Cancer Sci 2007; 98: 740–746)

Galectin-7 as a potential predictive marker of chemo-and/or radio-therapy resistance in oral squamous cell carcinoma

Treatment of advanced oral squamous cell carcinoma (OSCC) requires the integration of multimodal approaches. The aim of this study was to identify predictors of tumor sensitivity to preoperative radiotherapy/chemotherapy for OSCC in order to allow oncologists to determine optimum therapeutic strategies without the associated adverse effects. Here, the protein expression profiles of formalin‐fixed paraffin‐embedded (FFPE) tissue samples from 18 OSCC patients, termed learning cases, who received preoperative chemotherapy and/or radiotherapy followed by surgery were analyzed by quantitative proteomics and validated by immunohistochemistry in 68 test cases as well as in the 18 learning cases. We identified galectin‐7 as a potential predictive marker of chemotherapy and/or radiotherapy resistance, and the sensitivity and specificity of the galectin‐7 prediction score (G7PS) in predicting this resistance was of 96.0% and 39.5%, respectively, in the 68 test cases. The cumulative 5‐year disease‐specific survival rate was 75.2% in patients with resistant prediction using G7PS and 100% in patients with sensitive prediction. In vitro overexpression of galectin‐7 significantly decreased cell viability in OSCC cell line. Therefore, our findings suggest that galectin‐7 is a potential predictive marker of chemotherapy and/or radiotherapy resistance in patients with OSCC.

Gene expression analysis by oligonucleotide microarray in oral leukoplakia.

BACKGROUND Oral leukoplakias (LP) are the most frequent types of oral pre-cancerous lesions, but there is no accurate assessment of this malignant transformation or even genetic diagnosis of the oral epithelial dysplasia. We need to identify the new genetic diagnosis system of the epithelial dysplasia. METHODS Oligonucleotide microarray was used to analyze expression patterns of 29,952 genes in 10 LP patients. We compared the different gene expressions between mild dysplasia cases and severe dysplasia cases. RESULTS Ninety-six genes expressed differentially were selected as candidates for up-regulated in severe dysplasia. Subsequently, we further selected 16 genes with highest differentially expression. By hierarchical clustering analysis, the 10 cases were divided mild dysplasia from severe dysplasia. CONCLUSIONS The 16 genes are suggested as biomarker gene sets of efficacy and quickly recognized in the development of oral epithelial dysplasia.

Lack of Association between High-risk Human Papillomaviruses and Oral Squamous Cell Carcinoma in Young Japanese Patients

BACKGROUND Human papillomaviruses (HPV) may play an important role as one of the possible etiologies of oral squamous cell carcinoma (OSCC). The present study aimed to investigate the association between HPV and OSCC in young Japanese patients by examining the presence of HPV DNA and surrogate markers in OSCC tissues. MATERIALS AND METHODS Forty young patients with OSCC whose surgical specimens were available were analyzed and compared with 40 patients randomly recruited from a pool of patients aged >40 years. HPV DNA was detected using the polymerase chain reaction-based AMPLICOR(®) HPV test, and surrogate markers of HPV infection were analyzed using immunohistochemical techniques to detect p16(INK4a) and p53. RESULTS Only two (5%) young patients and one (2.5%) older patient were positive for HPV DNA. p16(INK4a) overexpression was identified in six (15%) young patients. p53 staining levels were not high in tissues of most young patients (27 patients, 67.5%). HPV DNA status did not significantly correlate with p16(INK4a) expression levels. Profiles of increased levels of p16(INK4a) expression with diminished levels of p53 staining were not associated with the presence of HPV DNA. The combined p53 with p16(INK4a) profiles were significantly correlated with alcohol consumption in younger patients (p=0.006). CONCLUSIONS RESULTS of the present study indicate that HPV is less likely to cause OSCC in young Japanese patients, and the p16(INK4a) expression level is not an appropriate surrogate marker for HPV infection in OSCC.

Clinical characteristics of multiple primary carcinomas of the oral cavity

OBJECTIVES This study aimed to clarify the clinical characteristics of multiple primary carcinomas of the oral cavity. MATERIALS AND METHODS We retrospectively reviewed the cases of 1015 patients who were treated during follow up for oral cancer at Tokyo Medical and Dental University between March 2001 and December 2012. We compared the clinical characteristics of 961 patients who developed single primary oral squamous cell carcinoma (SCC) during follow up and 54 patients who subsequently developed multiple primary carcinomas in the oral cavity. RESULTS Mean age at first diagnosis was significantly higher in patients with multiple primary carcinomas than single primary carcinoma. Multiple primary carcinomas showed a female predilection, were most prevalent in the gingiva, and tended to show earlier tumor and nodal stages than single primary carcinoma. The local recurrence rate was higher for multiple primary carcinomas than single primary carcinoma, and it increased with the number of multiple primary occurrences. The disease-specific survival rate at 10 years for patients with single primary carcinoma was 85.3% and that for patients with multiple primary carcinomas was 79.6%. The cumulative incidence rate for metachronous second multiple primary carcinomas after the onset of first carcinoma at 10 years was 8.0%. The recurrence of multiple primary carcinomas did not decrease the survival rate. CONCLUSION Differences were found in the clinical characteristics between patients with single oral SCC and those with multiple primary oral carcinomas. Early diagnosis and treatment as well as close long-term follow up are needed for patients with multiple primary oral carcinomas.

Functional outcomes and patient satisfaction after vascularized osteocutaneous scapula flap reconstruction of the mandible in patients with benign or cancerous tumours

Pre- and postoperative longitudinal assessment of oral functions including masticatory force, masticatory ability with solids, speech intelligibility, and subjective satisfaction was conducted for patients who underwent vascularized osteocutaneous scapula flap reconstruction after mandibulectomy. Postoperative changes in oral function and patient satisfaction were examined in relation to tumour type (i.e., cancerous vs. benign). A change in masticatory force was associated with a reduction in the number of occlusal support zones after surgery and clearly differed by tumour type. Despite all patients showing reduced masticatory force postoperatively, all were satisfied with the improvements in eating and chewing, which were greater in patients with benign tumours than in those with cancerous tumours. Although both groups of patients expressed satisfaction with the aesthetic appearance after surgery, patients with cancerous tumours were more satisfied. However, both groups showed decreased speech intelligibility scores and lower satisfaction with speech after surgery. Preoperative support in the form of reassurance and helping to alleviate anxiety about postoperative appearance would be beneficial for all patients, with additional support provided based on the specific tumour type.

Value of real-time tissue elastography for squamous cell carcinoma of the tongue

Objectives: Real-time tissue elastography is an ultrasound tomography technique that can objectively display tissue elasticity. The aim of this study was to evaluate the strain ratio of tongue carcinoma and the normal tongue and to investigate how clinico-pathological factors influenced the strain ratio. We also investigated differences in the strain ratio between tumor and non-tumor tissue after preoperative chemo-radiotherapy. Materials and methods: Patients with tongue carcinoma were categorized into two groups: 38 patients who underwent surgery alone and who underwent preoperative elastography (group 1), and 12 patients who underwent elastography before and after preoperative chemo-radiotherapy (group 2). Results: The mean strain ratios in group 1 were 0.16 ± 0.13 in tumor tissue and 0.82 ± 0.35 in normal tissue. Statistical analysis revealed that the strain ratio of tumor significantly correlated with tumor thickness. In group 2, the mean strain ratio of tumor tissue, scar tissue, and normal tissue were 0.19 ± 0.18, 0.51 ± 0.28, and 0.77 ± 0.58, respectively. The mean strain ratio of scar tissue was 0.51 and had lower elasticity than tumor tissue and higher elasticity than normal tissue. Conclusions: It was useful to characterize the tissue distortion using the strain ratio. Additional studies to establish a cutoff value for tumor tissue might help expand the indications for limited surgery. Correspondence to: Hiroyuki Harada, Professor, Oral and Maxillofacial Surgery, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan, Tel: +81-3-5803-5508, Fax: +81-3-5803-0199, E-mail: hiro-harada.osur@tmd.ac.jp Received: October 27, 2016; Accepted: November 04, 2016; Published: November 07, 2016 Introduction Preservation of function is important to preserve quality of life for patients undergoing treatment for oral cancer. The tongue is an essential organ for speech, food bolus formation and deglutition. Alternatively, this site is a good candidate for organ preservation techniques with the goal of preserving speech and swallowing function. Fortunately, preoperative chemo-radiotherapy has enabled many patients to avoid surgery and to improve cancer control with few side effects for patients with large unresectable cancers. These combined (chemo-radiotherapy) modality treatments lead to higher cure rates while maintaining the patient’s quality of life [1,2]. Indeed, advances in anticancer agent and radiotherapy have resulted in a potent antitumor effect in many patients. However, some patients treated with this strategy develop induration in the center of their tumor that histologically varies from viable tumor to scar tissue. If a determination could be made as to whether the remaining induration is tumor or scar tissue, it may be possible to establish criteria for limited resection versus no surgery [3-5]. Many diseases cause changes in the mechanical properties of tissues. The tissue flexibility is decreased by pathologic tissue changes, such as inflammation and tumor growth. Indeed cancer growth results in an increase in tissue stiffness, even during the early stages of cancer. Real-time tissue elastography is an ultrasound tomography modality that can objectively display tissue elasticity [6]. Basic principles including calculated method of strain ratio had been well described in the literature accompanied with a new guideline [7]. The strain distributions in tissues in response to external compression are closely related to the distribution of tissue elasticity. These strain images can give a clear illustration of underlying tissue stiffness and provides useful clinical information. Erastography is currently under investigation for diagnosing malignant breast tumor, thyroid tumor and cervical lymph node metastases [8,9]. However, there are no reports of its use in the evaluation of primary oral cancers. This method relies of the phenomenon in which compression of living tissue results in marked distortion of flexible tissues without affecting the shape of stiff tissues. When applying different levels of compression or relaxation, the elasticity imaging algorithm measures the displacement and strain by analyzing the RF (radiofrequency) signal in real time with subsequent conversion to a color-coded image. Using this method, the elasticity and strain ratio of squamous cell carcinoma of the tongue was measured, and the elasticity and strain ratio of the scar portion was quantified in cases in which preoperative therapy was performed [10]. Materials and methods Eligibility criteria The study was conducted between August 2011 and April 2015 in the Department of Oral and Maxillofacial Surgery of the Tokyo Medical and Dental University, Japan. All study protocols were approved by the institutional ethics committee (Approval number 729). All patients Aktar D (2016) Value of real-time tissue elastography for squamous cell carcinoma of the tongue Volume 2(6): 371-375 Dent Oral Craniofac Res, 2016 doi: 10.15761/DOCR.1000181 provided written informed consent before entry into this study. The subjects comprised 50 patients (38 men, 12 women; mean age, 59.1 years; age range, 30-86 years) who were treated for squamous cell carcinoma of tongue. Patients were categorized into two groups. Group 1 included 38 patients. These patients underwent surgery alone and elastography before glossectomy. Patients in group 1 underwent surgery for partial glossectomy (n=14) or for hemi-glossectomy (n=24). Group 2 included 12 patients who underwent elastography before and after preoperative chemo-radiotherapy. Patients in group 2 underwent surgery for hemiglossectomy (n=7) or for subtotal glossectomy (n=5). Preoperative therapies were chemoradiotherapy in 5 patients and chemotherapy in 7 patients. Preoperative chemoradiotherapy consisted of a total dose of S-1 at 65 mg/m2 per day for 5 consecutive days over 4 consecutive weeks with concomitant radiotherapy (40 Gy, provided at a fractional dose of 2 Gy for 5 days/week) in 4 patients. Preoperative chemotherapy consisted of a regimen of CDDP 80 mg/m2 (day 1) and 5-FU 800 mg/ m2 per day (days 1-5) plus a regimen of CDDP 60 mg/m2 and docetaxel 60 mg/m2 (day 1) and 5-FU 600 mg/m2 per day (days 1-5) for each one course plus 40 Gy of radiotherapy in one patient. Preoperative chemotherapy consisted of a regimen of CDDP 80 mg/m2 (day 1) and 5-FU 800 mg/m2 per day (days 1-5) for one course in 2 patient and for 2 courses in 2 patients. Preoperative chemotherapy consisted of a regimen of CDDP 60 mg/m2 and docetaxel 60 mg/m2 (day 1) and 5-FU 600 mg/m2 per day (days 1-5) for one course in one patient and for 2 courses in 2 patients. The median interval between end of preoperative treatment was 22.5 days (range, 13-35 days). T classification (group 1/ group 2) was T1: 6/0, T2: 26/4, T3: 6/8, and the N classification was N0: 30/3, N1: 4/1, N2b: 4/3 and N2c: 0/5. Histological classification of malignancy In group 1, the histological classification of malignancy was evaluated according to the multifactorial system proposed by Anneroth (Table 1) [11]. The histologic malignant score was determined using blocks of tissue that showed the maximum cross-sectional area. The number of mitoses was assessed by evaluating five random fields (magnification x400).

Primary Diffuse Large B-cell Lymphoma Arising in the Tongue Accompanied by Ataxia-telangiectasia: A Case Report.

Ataxia-telangiectasia (AT) is a rare autosomal recessive disorder that is characterized by progressive cerebellar ataxia, telangiectasia, immunodeficiency, and a predisposition to leukemia/lymphoma. Here we report a rare case of lymphoma of the tongue accompanied by AT. Tumour extirpation was performed and diffuse large B-cell lymphoma was diagnosed following pathologic examination. A whole-body survey showed no other enlarged lymph nodes or tumour. The female patient then received a modified dosage of COPAD (cyclophosphamide, vinblastine, pirarubicin, and prednisolone) plus rituximab to avoid severe complications. As of follow-up after 3 years and 5 months, she remains in complete remission. Patients showing AT need careful surveillance and long-term continuous follow-up.