Tomoki Sumida

Histological study on pN upgrading of oral cancer

Abstract The International Union Against Cancer (UICC) does not define the number of sections required from each regional lymph node to record pTNM classification. This study was designed to clarify the incidence of occult metastasis and to assess the pN upgrading of patients with oral cancer. Ultimately, this study led to a proposal for appropriate semiserial sectioning guidelines. Five hundred fifty-four nonmetastatic cervical lymph nodes taken from 73 patients with oral cancer were subjected to hematoxylin-eosin (HE) staining and keratin immunohistochemistry. Micrometastases, defined as foci ≤3 mm, were detected in 29 sites of 23 lymph nodes (4.2%) of 16 patients (21.9%). In 9 patients (12.3%) pN upgrading was needed: in 6 from pN0 to pN1, in 1 from pN0 to pN2b, and in 2 from pN1 to pN2b. The remaining 13 lymph nodes with occult metastasis were found in 5 pN2b and 2 pN2c patients, resulting in no pN upgrading. Occult metastasis was also detected in 6 small lymph nodes ≤5 mm in diameter. The average minor axis of the micrometastasis was 1.36±0.85 mm. We propose that the lymph nodes should be cut and examined at 1-mm intervals to detect micrometastatic foci and to evaluate the pN classification accurately.

Genetic diagnosis of micrometastasis based on SCC antigen mRNA in cervical lymph nodes of head and neck cancer

This study is designed to assess gene expression of squamous cell carcinoma antigen (SCCA) mRNA to detect micrometastases in cervical lymph nodes (LNs) of head and neck cancer. We examined the expression of SCCA mRNA in 12 primary tumors and 212 cervical LNs (101 LNs taken from 8 patients with tongue cancer, 71 from 7 patients with gingival cancer, 19 from 2 patients with laryngeal cancer, 9 from 2 patients with pharyngeal cancer, 7 from 1 patient with cancer of the buccal mucosa, and 5 from 1 patient with cancer of floor of the mouth). Detectability of metastatic LNs by nested and single reverse transcriptase-polymerase chain reaction (RT-PCR) was compared with semiserial sections (hematoxylin-eosin staining and keratin immunostaining). All primary tumors expressed SCCA mRNA. Of 198 histologically metastasis-negative nodes, SCCA mRNA was detected in 37 (18.7%) by nested PCR. Eleven micrometastatic foci in 9 LNs (4.6%) were discovered by semiserial sectioning. This suggests that SCCA mRNA is a promising tumor marker for detecting the micrometastases in cervical LNs of head and neck cancer.

BASIC EVIDENCE OF MOLECULAR TARGETED THERAPY FOR ORAL CANCER AND SALIVARY GLAND CANCER

Recently, attention has been focused on molecular targeted cancer therapy in various tumors. Although there is no single consistent molecular target specific for oral squamous cell carcinoma (OSCC) and salivary gland cancer (SGC), there are a number of promising candidate proteins. The aim of this review is to introduce the basic evidences to support the molecular targeting for OSCC and SGC.

Telomerase components as a diagnostic tool in human oral lesions

Telomerase activity is considered to be a diagnostic marker of malignancy since most malignant cells express this activity and most somatic cells do not. However, the detection of telomerase activity is rather complicated and is affected by many factors. Recently, human telomerase components were cloned and found to consist of 3 subunits. We assessed which component of telomerase best correlates with malignancy in order to study the possibilities for developing a new diagnostic marker. Telomerase activity was measured by a telomeric repeat amplification protocol (TRAP) assay, and the telomerase components hTR, hTRT-mRNA and TP1-mRNA were detected by the reverse transcriptase-polymerase chain reaction (RT-PCR). Twenty-five of 26 oral malignant lesions, 9 of 22 benign lesions and none of 19 normal control tissues exhibited distinct telomerase activity. hTR and TP1-mRNA expression levels were detected in all malignant lesions and normal control tissues and had no significant correlation with the telomerase activity results. In contrast, hTRT-mRNA expression was closely associated with telomerase activity. All lesions expressing hTRT were telomerase positive. In addition, some samples of dysplastic lesions, benign tumors, lichen planus and normal mucosa exhibiting poor telomerase activity revealed weak expression of hTRT. Expression levels of hTRT-mRNA positively correlated with clinical and pathological findings. Detection of hTRT-mRNA by RT-PCR appeared to be more sensitive for telomerase than measurement of telomerase activity by the TRAP assay. Detection of hTRT-mRNA may provide information useful in the diagnosis of oral malignancies.

Keratin mRNA for detecting micrometastasis in cervical lymph nodes of oral cancer

We studied three keratin (K) gene candidates, K13, K19, and K20 mRNAs, for detecting micrometastases in cervical lymph nodes (LNs) by reverse transcriptase-polymerase chain reaction (RT-PCR). Of 166 histologically metastasis-negative nodes, 24 micrometastatic LNs (14. 4%) were detected based on K13 gene expression. Keratin 19 mRNA is an inadequate marker for the genetic diagnosis due to not only illegitimate gene expression from lymphatic tissue but also gene expression from the ectopic salivary gland. Keratin 20 mRNA showed low sensitivity. It is suggested that K13 mRNA may be a promising tumor marker among these keratin genes for detecting the micrometastases in cervical LNs of oral cancer.

Infiltrating angiolipoma of the cheek: a case report and a review of the literature.

Lipoma of the oral cavity is a common benign soft tumor. However, angiolipoma, showing characteristics of both lipoma and hemangioma, usually develop in the trunk and extremities, and rarely arise in the head and neck region. The occurrence of angiolipoma in the cheek is very low; to our knowledge, 5 cases have been reported in the English literature. 2-5 No previous case showed an infiltrating mass. We report a case of angiolipoma that broadly infiltrated the cheek and review the features of angiolipomas arising in the oral cavity.

Enhancement of radiosensitivity in head and neck cancer cells by ZD1839 ('IRESSA'), a selective epidermal growth factor receptor tyrosine kinase inhibitor.

Overexpression of epidermal growth factor receptor (EGFR) is frequently observed in many solid tumor types, including head and neck squamous cell carcinomas (HNSCC). Recent laboratory experiments have demonstrated that high EGFR levels correlate with increased tumor resistance to radiation. This study investigated the relationship between EGFR expression levels and radiosensitivity in 5 HNSCC cell lines (HSC2, HSC3, HSC4, SCC25, and Ca9–22) and whether treatment with ZD1839 (‘Iressa’), a selective EGFR-tyrosine kinase inhibitor (TKI), would improve tumor cell response to radiotherapy. ZD1839 suppressed the growth of HNSCC cell lines in a dose- and time-dependent manner. Radiosensitivity of these HNSCC cell lines, assessed by a clonogenic survival assay, differed greatly and the expression of EGFR varied. EGFR expression levels (EGFR numbers/cell) correlated with increased tumor resistance to radiation (f[x]= 4.54 X, R2 = 0.715; f[x]: EGFR numbers/cell, X: radiosensitivity; D10). Following exposure of the HNSCC cells to 1.0 &mgr;M ZD1839 and radiation (0–10 Gy), greater than additive growth inhibitory effects were observed. These results suggest that ZD1839 could enhance tumor radiosensitivity and inhibit tumor growth after radiation, indicating that this combination could have clinical potential in the treatment of patients with head and neck cancer.

Serum autoantibody to sideroflexin 3 as a novel tumor marker for oral squamous cell carcinoma

The purpose of this study is to establish a tumor marker that can be applied for the early detection and follow‐up of oral cancer patients. Employing the proteomic approach using MALDI TOF‐MS, 2‐DE, patients sera and culturing cell lines, the serum autoantibodies (autoAbs) were screened and the serum levels were estimated by ELISA. Targeting the tumor cell invasion into the surrounding stromal tissues, MRC‐5 human fibroblasts were employed as the target cells and a mitochondrial membrane protein, sideroflexin 3 (SFXN3), was identified. The serum anti‐SFXN3‐autoAb levels elevated in patients with the oral squamous cell carcinoma significantly: with 77% sensitivity and 89% specificity against control samples. The serum anti‐SFXN3‐autoAb levels were mildly correlated with the primary tumor sizes, however, the levels were slightly highly elevated in T1 early cancer. An immunohistochemical analysis revealed that the SFXN3 protein is expressed in the stromal fibroblasts between the caner nests and also in the basal layer of the squamous epithelium. Changes in the serum anti‐SFXN3‐autoAb levels after therapy correlated with the clinical tumor burden. These findings demonstrated that the serum anti‐SFXN3‐autoAb is worthy of clinical evaluation as a potentially of the novel tumor maker for the early detection of oral squamous cell carcinoma.

Custom-made titanium devices as membranes for bone augmentation in implant treatment: Modeling accuracy of titanium products constructed with selective laser melting

OBJECTIVE The purpose of this study was to verify the modeling accuracy of various products, and to produce custom-made devices for bone augmentation in individual patients requiring implantation. MATERIALS AND METHODS Two-(2D) and three-dimensional (3D) specimens and custom-made devices that were designed as membranes for guided bone regeneration (GBR) were produced using a computer-aided design (CAD) and rapid prototyping (RP) method. The CAD design was produced using a 3D printing machine and selective laser melting (SLM) with pure titanium (Ti) powder. The modeling accuracy was evaluated with regard to: the dimensional accuracy of the 2D and 3D specimens; the accuracy of pore structure of the 2D specimens; the accuracy of porosity of the 3D specimens; and the error between CAD design and the scanned real product by overlapped images. RESULTS The accuracy of the 2D and 3D specimens indicated precise results in various parameters, which were tolerant in ISO 2768-1. The error of overlapped images between the CAD and scanned data indicated that accuracy was sufficient for GBR. In integrating area of all devices, the maximum and average error were 292 and 139 μm, respectively. CONCLUSIONS High modeling accuracy can be achieved in various products using the CAD/RP-SLM method. These results suggest the possibility of clinical applications.

Silencing Id-1 inhibits lymphangiogenesis through down-regulation of VEGF-C in oral squamous cell carcinoma

Our previous study demonstrated that overexpression of Id-1 (inhibitor of differentiation/DNA binding) was associated with lymphatic metastasis in human oral squamous cell carcinoma (OSCC). In this study, we further unveiled the association of Id-1 with vascular endothelial growth factor-C (VEGF-C) and peritumoral lymphatic vessel density (PLVD), and the effect of silencing Id-1 on inhibiting lymphangiogenesis in OSCC. We found that Id-1 was associated with VEGF-C (r=0.569, p<0.001) and PLVD (r=0.240, p<0.001) in OSCC. Lentivirus-mediated RNA interference targeting Id-1 in an OSCC cell line Tca8113 resulted in down-regulation of VEGF-C (p=0.003, 0.007). Moreover, when Id-1 was suppressed by injecting Id-1-siRNA-lentivirus into the transplanted tumors in nude mice, VEGF-C was down-regulated (p=0.018) and the PLVD decreased (p=0.001). Our results suggest that Id-1 was correlated with lymphangiogenesis in OSCC. Silencing Id-1 could inhibit lymphangiogenesis through down-regulation of VEGF-C and it might be a promising treatment modality for the lymphatic metastasis of OSCC.