Fibroblast VEGF-receptor 1 expression as molecular target in periodontitis.

AIM: Degradation of extracellular matrices is an integral part in periodontitis. For antagonizing this pathophysiological mechanism, we aimed at identifying gene expression profiles in disease progression contributing periodontitis-associated fibroblasts (PAFs) versus normal gingival fibroblasts to determine their molecular repertoire, and exploit it for therapeutic intervention. MATERIALS AND METHODS: Applying an exploratory analysis using a small number of microarrays in combination with a three dimensional (3D) in vitro culture model that incorporates some aspects of periodontitis, PAFs were initially characterized by gene-expression analyses, followed by targeted gene down-regulation and pharmacological intervention in vitro. Further, immunohistochemistry was applied for phosphorylation analyses in tissue specimens. RESULTS: PAFs were characterized by 42 genes being commonly up-regulated >1.5-fold, and by five genes that were concordantly down-regulated (<0.7-fold). Expression of vascular endothelial growth factor (VEGF)-receptor 1 (Flt-1) was highly enhanced, and was thus further explored in in vitro culture models of periodontal fibroblasts without accounting for the microbiome. Phosphorylation of the VEGF-receptor 1 was enhanced in PAFs. Receptor inhibition by a specific VEGF-receptor inhibitor or intrinsic down-regulation by RNAi of the VEGF-receptor kinase in 3D gel cultures resulted in significant reduction in collagen degradation associated with increased tissue inhibitor of metalloproteinase expression, suggesting that Flt-1 may contribute to periodontitis. CONCLUSION: Based on the finding that VEGF-receptor kinase inhibition impaired collagen degradation pathways, Flt-1 may represent a candidate for therapeutic approaches in periodontitis.

Retrospective clinical study of 296 patients with mass lesions of the tongue.

To better understand the clinical features of mass lesions of the tongue, we retrospectively evaluated frequency, recurrence rate, and complications in 296 patients who had undergone surgery for such lesions. The diagnoses were fibroma (43.6%), mucous cyst (14.2%), papilloma (11.8%), hemangioma (7.8%), granuloma (6.4%), lipoma (1.4%), schwannoma (1.0%), ectopic tonsil (0.7%), and other (13.2%). Recurrence was noted in two patients (0.7%). Twenty-two patients (7.4%) developed surgical complications, including lingual nerve paralysis (6.4%), glossodynia (0.6%), and postoperative infection (0.3%). Lingual nerve paralysis was observed in the ventral portion (42.1%) of the tongue, apex (36.8%), lateral border (10.5%), and dorsum (10.5%). When all sites were considered together, there was no significant difference in the number of patients presenting with lingual nerve paralysis (P = 0.075). However, there were significant differences in lingual nerve paralysis at the lateral border (P < 0.05), apex (P < 0.05), and dorsum (P < 0.001) but not at the ventral portion (P > 0.05) in the size of the patients with versus without it which suggests that the risk of lingual nerve paralysis is higher at the ventral tongue, regardless of tumor size. These results shed light on the clinical features of mass lesions of the tongue.

Butyrate inhibits oral cancer cell proliferation and regulates expression of secretory phospholipase A2-X and COX-2.

Although surgical resection is the first choice for oral cancer, the development of new anti-cancer drugs is of great interest. The effect of the histone deacetylation inhibitor, sodium butyrate (NaBu) on oral cancer cell (OCC) HSC-3 and HSC-4 proliferation in vitro was investigated. The synthesis of rate-limiting enzymes such as sPLA2 (-IIA, -V, -X) and COX-2 was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, as well as PGE2 by ELISA. NaBu acted in a concentration-dependent manner. Over 3 mM, it inhibited OCC proliferation, due to increased p21 expression and cell cycle arrest in the G2/M-phase. At low concentration (< or = 1 mM), NaBu showed no effects or enhanced cell proliferation. NaBu also regulated COX-2 and sPLA2-X expression, and augmented PGE2 synthesis in OCC. These results indicate that NaBu is a novel candidate agent for the treatment of oral cancer. The treatment efficacy must be investigated in additional experiments considering NaBu concentration and tumor cell heterogeneity.

Rapid and Specific Screening Assay for KRAS Oncogene Mutation by a Novel Gene Amplification Method.

BACKGROUND: Epidermal growth factor receptor (EGFR) is a target of molecular therapeutics for colorectal cancer. However, mutation of Kirsten rat sarcoma viral oncogene homolog (KRAS) gene at codons 12 and 13 attenuates the therapeutic effect of anti-EGFR therapies. Therefore, the detection of KRAS gene mutation is important for therapeutic decision-making. MATERIALS AND METHODS: KRAS gene mutation at codons 12 (c.34G>T, c.35G>C, c.35G>A) and 13 (c.38G>A) in six cancer cell lines were investigated. A loop-mediated isothermal amplification-based procedure was developed that employed peptide nucleic acid to suppress amplification of the wild-type allele. RESULTS: This mutation-oriented gene-amplification procedure can amplify the DNA fragment of the KRAS gene with codon 12 and codon 13 mutation within 30 min. Moreover, boiled cells can work as template resources. CONCLUSION: This newly developed procedure can be useful for patient stratification for anti-EGFR therapies.

Unusual cyst-like lesions in the parapharyngeal space associated with recurrence of tongue carcinoma.

A 54-year-old male presented with the complaint of a painful sore on the left side of his tongue. Our examination found an ulcer 15 x 20 mm in size on the left edge of the tongue, with peripheral indurations. The lesion was diagnosed histopathologically as squamous cell carcinoma (T2N0M0). Consequently, the lesion was surgical removed and radical neck dissection was performed. Four months after the operation, two unusual cyst-like lesions were identified in the parapharyngeal space by CT and MRI. A biopsy specimen revealed recurrent carcinoma with a cyst-like structure. The route of the tumor metastasis into the parapharyngeal space was obscured, but it was speculated that the excessive lymph accumulation was due to a lymphatic occlusion caused by the surgical procedure, proliferation of the metastatic carcinoma, or stagnation and accumulation of tissue fluid caused by parapharyngeal invasion by the recurrent lesion.

Postoperative mental disorder following prolonged oral surgery.

A prolonged period of oral surgery is a potential risk factor of postoperative mental disorders although no such report has been published to date. We retrospectively studied perioperative features in 36 patients who underwent prolonged (10 hours or more) of oral surgery. Patients were categorized as pre-delirium (Pre-D) when they manifested 1 or 2 symptoms and delirium (D) when they showed more than 2 symptoms, according to the modified International Classification of Diseases, 10th edition. Of the 36 patients who returned to a normal mental state without drug therapy, 13 were classified as D and 14 were Pre-D. A number of patients had moderate complications preoperatively, and massive hemorrhaging occurred during the operation in some Pre-D and D patients. Age was greater in D (62.0 +/- 9.9 years) than in Pre-D (56.0 +/- 13.8 years) patients. Propofol-based general anesthesia was most commonly employed. The time prior to appearance of pre-delirium was significantly shorter in D (30.0 +/- 16.7 hours) than in Pre-D (55.0 +/- 35.0 hours) group patients. Our results indicate that, in general, patients predisposed to postoperative mental disorders have moderate complications preoperatively, are generally older than 50-years-old, receive propofol-based general anesthesia and/or experience a massive hemorrhage during the operation.

Prognostic value of LYVE-1-positive lymphatic vessel in tongue squamous cell carcinomas.

The density of lymphatic vessels in 52 cases of human tongue squamous cell carcinoma (TSCC) and normal portions was analyzed. TSCC specimens were immunostained with antibodies against lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1) and podoplanin monoclonal antibody (D2-40). The significance of the LYVE-1-positive vessel density (LVD) was calculated in 6 topographic areas and investigated on the basis of specific clinical and histo-pathological parameters. LYVE-1 positivity was more evident in the muscular area than the submucosal area, while small D2-40-positive lymphatic vessels were not demonstrable in muscular endomysium. The LVD in peri-tumoral submucosal and peri-tumoral muscular areas was lower than in normal counterparts (p<0.01). LVD was higher in the tumor invasion front as compared to tumor-associated stroma (p<0.01). Low LVD in invasion front and peri-tumoral submucosal area was significantly related to regional lymph node metastasis (p<0.05 and p<0.01, respectively). The decrease of LYVE-1-positive lymphatic vessels in the invasion front and peri-tumoral submucosal area would seem to predict cervical lymph node metastasis in TSCC.

Anaplastic large cell lymphoma in gingiva: case report and literature review.

We report an unusual case of gingival anaplastic large-cell lymphoma (ALCL) that occurred in a 76-year-old Japanese woman who showed marked gingival swelling in both the maxilla and mandible. Although the patient received caries and periodontal therapy at the outpatient clinic of our dental hospital, the gingival swelling remained and she was referred to the oral surgery department, where a biopsy of the gingiva was performed. The specimens showed proliferation of large atypical and amphophilic epithelioid cells beneath the covering epithelium. Immunohistochemical analysis of the proliferating cells revealed positivity for CD30 and T-cell markers, such as CD45RB, as well as CD45RO antibodies, and they were weakly positive for the granzyme B antibody. In contrast, the tumor cells were negative for all B-cell markers as well as for CD3, CD56, S-100 protein, epithelial membrane antigen, and p80(NPM/ALK) antibodies. Based on the clinical and histopathologic features, the lesion was diagnosed as an ALCL in both the upper and the lower gingiva. This is an extremely rare case, in which a specific subtype of T-cell lymphoma appeared in the oral cavity.