Kenichiro Uchida

Comparative genomic hybridization reveals genetic progression of oral squamous cell carcinoma from dysplasia via two different tumourigenic pathways

To clarify the genetic pathway(s) involved in the development and progression of oral squamous cell carcinoma (OSCC), as well as the relationship between genetic aberrations and biological characteristics of OSCC tumours, comparative genomic hybridization was used to analyse genetic alterations in both primary OSCCs and adjacent dysplastic lesions of the same biopsy specimens from 35 patients. Gain of 8q22–23 was the most frequent alteration in both OSCC and mild dysplasia, and was considered the earliest event in the process of oral tumourigenesis. The average number of DNA sequence copy number aberrations (DSCNAs) increased with progression from mild dysplasia to invasive carcinoma (r = 0.737, n = 70, p < 0.001). OSCC samples were classified as having a large or small number of DSCNAs (OSCC‐L, 21.4 ± 4.7 DSCNAs or OSCC‐S, 10.0 ± 1.7 DSCNAs, respectively; p < 0.0001). Gains of 3q26‐qter, 8q, 11q13, 14q, and 20q and losses of 4q, 5q12–22, 6q, 8p, 13q, and 18q22‐qter were common to OSCC‐L and OSCC‐S. Gains of 5p15, 7p, 17q11–22, and 18p and losses of 3p14–21, 4p, and 9p were detected exclusively in OSCC‐L. The average number of DSCNAs depended on whether the samples showed OSCC‐ L or dysplasia plus OSCC‐L, or showed OSCC‐S or dysplasia plus OSCC‐S (p = 0.001). Gain of 5p15 and losses of 4p and 9p were detected even in dysplastic lesions adjacent to OSCC‐L samples. Loss of 4p was associated with node metastasis by multivariate analysis (p = 0.013). OSCC‐L tumours were more often T3–T4 stage tumours than T1–T2 stage tumours (p = 0.03). These findings suggest that two different types of OSCC, OSCC‐L associated with high‐stage cancer and OSCC‐S associated with low‐stage cancer, arise from different types of dysplasia via different genetic pathways. Copyright

Interaction of OGG1 Ser326Cys polymorphism with cigarette smoking in head and neck squamous cell carcinoma.

Recent molecular epidemiological studies have demonstrated that the human oxoguanine glycosylase 1 (OGG1) gene polymorphism may be associated with various cancers. To determine whether the OGG1 Ser326Cys polymorphism interacts with clinicopathological parameters including smoking and alcohol intake in head and neck squamous cell carcinoma (HNSCC), DNA samples from 192 patients with primary HNSCC were genotyped and studied by the case‐only design. We observed an association between the Cys/Cys genotype and HNSCC with cigarette smoking of more than 40 pack‐years by a multivariate logistic regression analysis (OR = 8.10, 95% CI = 1.06–61.73). No significant association of this genotype with alcohol intake was observed. Our present data suggest a possible interaction between the OGG1 Ser326Cys polymorphism and smoking in HNSCC.

Loss of 3p26.3 is an independent prognostic factor in patients with oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is a common malignancy worldwide and the prognosis for patients with advanced-stage OSCC is particularly poor. To identify DNA copy number aberrations and candidate genes associated with a poor or favorable outcome, we analyzed the genome profiles of OSCC tumors by array-based comparative genomic hybrid-ization (A-CGH). This technique uses DNA microarray technology to detect genomic copy number variations at a higher resolution level than chromosome-based CGH. Fifty patients with primary OSCCs were included in the study. Of these 50 patients, 37 were treated surgically and 13 were treated without surgery and had received irradiation and/or chemotherapy. All samples were analyzed by A-CGH. Gains were detected frequently (>50%) at chromosomal regions 5p15.33, 7p22.3, 8q21.1-24.3, 9q34.3, 11q13, 16p13.3 and 20q13.3. Losses were frequently detected at 3p22, 3p14 and 4q35.2. High-level gains were recurrently (>10%) detected at each of 5p15, 7p22, 7p11, 8q24, 11q13, 11q22 and 22q11. Gains of 2p25.1, 11p15, 16p13.3, 16q24.3 and 20q13.3 were inversely correlated with nodal metastasis. In 37 of the 50 OSCC patients treated with surgery, gains of 8q12.1-24.22 and losses of 3p26.2-3 were associated with disease-specific survival (p<0.01). Loss of a 0.2 Mb chromosomal region in 3p26.3 was associated with a poor prognostic outcome in the Kaplan-Meier analysis (p<0.01 by the log-rank test). Multivariate analysis revealed that loss of 3p26.3 is an independent prognostic factor (p<0.01) of OSCC. Loss of a 0.2 Mb chromosomal region in 3p26.3 including the CHL1 (cell adhesion molecule with homology to L1CAM1) gene was identified as a novel potential marker for predicting the prognosis of patients with OSCC.

ALDH2 1510 G/A (Glu487Lys) polymorphism interaction with age in head and neck squamous cell carcinoma.

Recent molecular epidemiological studies have revealed a possible association of the acetaldehyde dehydrogenase-2 (ALDH2) 1510 G/A (Glu487Lys) polymorphism with various cancers including head and neck squamous cell carcinoma (HNSCC). To further elucidate the significance of this polymorphism in HNSCC development, the relationship between ALDH2 1510 G/A and clinicopathological parameters, cigarette smoking or alcohol intake was evaluated in patients with HNSCC. DNA samples from 192 patients with primary HNSCC and 192 age- and gender-matched healthy controls were genotyped and statistically evaluated. Although there was no significant difference in the genotype distribution of ALDH2 1510 G/A between cases and controls, we found that the frequency of the ALDH2 genotypes with the mutated A (Lys) allele was greater in patients aged <66 years than in those aged ≧66 years (p = 0.034). This tendency became more evident in patients with the habit of drinking (n = 143; p = 0.009). The association of ALDH2 1510 G/A with age remained significant after multivariate logistic regression analysis was performed for the patients (odds ratio for an age interval for 1 year, 0.970; 95% confidence interval, 0.943–0.998). The present data suggest a possible interaction between the ALDH2 1510 G/A polymorphism and age in HNSCC.

Proteomic profiling of differential display analysis for human oral squamous cell carcinoma: 14-3-3 σ Protein is upregulated in human oral squamous cell carcinoma and dependent on the differentiation level

Oral squamous cell carcinoma (OSCC) has an absolute majority of all oral cancer. We used proteomic technology to analyze the protein expression profile in OSCC tissues and accompanying surrounding normal tissues in four oral locations (buccal mucosa, gingival mucosa, oral floor, and tongue). Ten protein spots were overexpressed more strongly in cancer tissues than normal ones, and were identified as proliferating cell nuclear antigen, 14‐3‐3 ε, 14‐3‐3 σ, proteasome subunit α type 5, translationally controlled tumor protein, eukaryotic translation initiation factor 3 subunit, macrophage capping protein, and mitochondrial isocitrate dehydrogenase subunit α. Macrophage capping protein and mitochondrial isocitrate dehydrogenase subunit α had two spots. Especially, we focused on 14‐3‐3 σ protein, one of the eight identified proteins, and assessed its expression level in four oral locations of OSCC by using differential display methods. The expression level of 14‐3‐3 σ protein was upregulated in four locations of oral cavity. Eight proteins which we identified in this study may play an important role in OSCC carcinogenesis and progression and could be used as diagnostic biomarkers of OSCC.

Screening for DNA copy number aberrations in mucinous adenocarcinoma arising from the minor salivary gland: two case reports.

Mucinous adenocarcinoma (MAC) is a rare malignancy in the minor salivary gland. To our knowledge, genomic alterations in this tumor have not been reported previously. To identify DNA copy number aberrations, we applied comparative genomic hybridization (CGH) to four samples of MAC in minor salivary gland derived from two patients: a primary tumor and two cervical metastatic lymph nodes from one patient, and a primary tumor from the other patient. Copy number increases were commonly detected in 1q21∼q31 and 20q13, and these may play an important role in MAC carcinogenesis. Copy number increases in 1q, 12p, 12q, and 20q were commonly detected in all three samples derived from patient 1, and gain of 7p and loss of chromosome 4 were additionally detected in the two samples derived from metastatic lymph nodes. Amplifications were also detected in the chromosomal regions 8q22∼qter, 12p11∼p12, 12q11∼q21, and 20q13. Amplification of MDM2 (12q15) and of AURKA (20q13) was detected with fluorescence in situ hybridization. The DNA copy number aberrations detected in MAC in minor salivary glands were different from those reported for colorectal MAC. The present findings are novel in identifying genomic alterations of MAC arising from the minor salivary gland.

Central Adenoid Cystic Carcinoma of the Mandible With Multiple Bone Metastases: Case Report

ndodontic or restorative procedure. In addition, postperative tooth hypersensitivity induced by the odonectomy should be taken into consideration as a potenial complication. This case report describes a novel pproach to the extraction of horizontally impacted andibular third molars that have a high risk of IAN aresthesia. This staged approach may be promising, nd further investigations may be warranted to evaluate ts efficacy in a large sample of patients.

Loss of 6q or 8p23 is associated with the total number of DNA copy number aberrations in adenoid cystic carcinoma

We analyzed 10 adenoid cystic carcinomas (ACCs) of the salivary glands by array-based comparative genomic hybridization (a-CGH) using DNA chips spotted with 4,030 bacterial artificial chromosome clones. After the data smoothing procedure was applied, a total of 88 DNA copy number aberrations (DCNAs) were detected. The frequent (≥30%) DCNAs were loss of 6q23-27 and 8p23, and gains of 6p, 6q23, 8p23 and 22q13. High-level gains were detected on 12q15, including MDM2 in two cases. These two cases showed an immunohistochemically high-level (>50%) expression of MDM2 and a low-level expression of p53 (<20%). Furthermore, the total number of DCNAs was significantly greater in ACCs with loss of 6q compared to other ACCs, and in ACCs without the loss of 8p23 compared to other ACCs, respectively. Although limitations exist, a-CGH detected several candidate chromosomal imbalances associated with accumulation of DCNAs in ACCs.

Variant form of stiff-man syndrome with neck pain: report of a case treated with muscle afferent block

continuous discharges at rest in the trapezius, deltoid, biceps, and triceps muscles (Fig. 1a). After obtaining informed consent, we performed MAB on the trapezius muscle, which were the site of severe pain. Bupivacaine (0.125%, 10 ml) was injected into the muscle (MAB with local anesthetics) twice a week. The pain and rigidity of the trapezius muscle were relieved for 2 to 5 h after the block, while voluntary muscle power was preserved. We therefore commenced MAB with alcohol, which has been shown effective for dystonia [2]. The techniques were as follows. The trapezius muscle was identified (the involuntary contracting muscle is identified with echo), and 8 ml of 1% mepivacaine was injected into the muscle belly using a 25-G, 25-mm needle, over 30s. This was followed by injection of 1 ml of absolute alcohol over 10 s and then injection of 2ml of 1% mepivacaine over 10 s. Mepivacaine given before and after alcohol injection is helpful to prevent severe pain from alcohol injection and from possible leakage during withdrawal of the needle. The block was repeated twice a week, mainly on the left side, and a few times on the right side. The symptoms were substantially improved. Induration at the injection site was the only complication. MAB was performed a total of 40 times, using either bupivacaine alone (30 times) or alcohol in combination with mepivacaine (10 times), resulting in a decrease in pain from 83 mm to 39mm on a visual analogue scale. Sustained rigidity in the trapezius muscle disappeared, and rigidity was observed only intermittently. Rigidity of the trapezius muscle, which had been observed in the entire muscle, was now confined to a smaller area, and some trigger points became apparent. Although involuntary movement of the muscles continued, pain became controllable with trigger point block once every 2 weeks. Surface electromyograms after the latest MAB showed minimum discharges in the trapezius muscles.