Transcriptome of Oral Cancer Cells Adapted to Suspension Culture Is Potentially Related to Cancer Progressive Phenotypes.

BACKGROUND/AIM: Cervical lymph node metastasis worsens oral cancer prognosis. Cancer cells with high metastatic ability can delay or resist apoptosis and survive in the floating condition during circulation. The involved genes and pathways in this process remain largely unknown. This study aimed to establish an oral cancer cell line adapted to suspension culture by in vitro selection and perform gene expression analysis. MATERIALS AND METHODS: The oral cancer cell subline adapted to suspension culture was isolated by in vitro selection from the oral cancer cell line, HSC-3. The transcriptome profiles of HSC-3 and its subline were compared using gene expression microarrays. Gene Ontology (GO) enrichment analysis, Gene Set Enrichment Analysis (GSEA), and Ingenuity Pathway Analysis (IPA) were performed to predict the involved pathways and molecules in cancer progression. RESULTS: The subline was designated as HSC-3S5 The cellular viability of HSC-3S5 cells at the suspension culture was higher than that of HSC-3 cells. A total of 961 genes were differentially expressed between HSC-3 and HSC-3S5 cells under the threshold cut-off (FDR-adjusted p-value of <0.05 and absolute fold change of >1.5). GO terms, such as growth regulation, were enriched in the DEGs. GSEA revealed the association between the DEGs and significant gene sets, including metastasis and stemness. IPA predicted that the proliferation-related pathways were enhanced while the apoptotic pathway was inhibited in HSC-3S5 cells compared to HSC-3 cells. CONCLUSION: Our transcriptome analysis revealed several potentially activated pathways and molecules in the floating-adapted oral cancer cells and indicated molecular implications for cancer progression.

Cytomorphology and Gene Expression Signatures of Anchorage-independent Aggregations of Oral Cancer Cells.

BACKGROUND/AIM: Cancer cells with high anchorage independence can survive and proliferate in the absence of adhesion to the extracellular matrix. Under anchorage-independent conditions, cancer cells adhere to each other and form aggregates to overcome various stresses. In this study, we investigated the cytomorphology and gene expression signatures of oral cancer cell aggregates. MATERIALS AND METHODS: Two oral cancer-derived cell lines, SAS and HSC-3 cells, were cultured in a low-attachment plate and their cytomorphologies were observed. The transcriptome between attached and detached SAS cells was examined using gene expression microarrays. Subsequently, gene enrichment analysis and Ingenuity Pathway Analysis were performed. Gene expression changes under attached, detached, and re-attached conditions were measured via RT-qPCR. RESULTS: While SAS cells formed multiple round-shaped aggregates, HSC-3 cells, which had lower anchorage independence, did not form aggregates efficiently. Each SAS cell in the aggregate was linked by desmosomes and tight junctions. Comparative transcriptomic analysis revealed 1,698 differentially expressed genes (DEGs) between attached and detached SAS cells. The DEGs were associated with various functions and processes, including cell adhesion. Moreover, under the detached condition, the expression of some epithelial genes (DSC3, DSP, CLDN1 and OCLN) were up-regulated. The changes in both cytomorphology and epithelial gene expression under the detached condition overall returned to their original ones when cells re-attached. CONCLUSION: The results suggest specific cytomorphological and gene expression changes in oral cancer cell aggregates. Our findings provide insights into the mechanisms underlying anchorage-independent oral cancer cell aggregation and reveal previously unknown potential diagnostic and therapeutic molecules.

Sentinel lymph node biopsy with a handheld cordless magnetic probe following preoperative MR lymphography using superparamagnetic iron oxide for clinically N0 early oral cancer: A feasibility study.

PURPOSE: Radioisotope (RI) tracers are generally used for preoperative mapping of sentinel lymph node (SLN) and intraoperative detection with a portable γ probe. However, the use of RI has several limitations. Therefore, a method without RI is required for the widespread application of SLN biopsy. The purpose of this study was to evaluate the feasibility of SLN biopsy with a handheld cordless magnetic probe following magnetic resonance lymphography (MRL) using superparamagnetic iron oxide (SPIO) and for clinically N0 early oral cancer. MATERIALS AND METHODS: MRL using SPIO and SLNB with the handheld cordless magnetic probe were performed for 27 patients with clinically N0 early oral cancer. RESULTS: In all 27 patients (100%), SLNs were detected by MRL, and the total and mean number of SLNs were 73 and 2.7, respectively. All SLNs identified by MRL were detectable using the magnetic probe in all patients. CONCLUSIONS: SLNB with handheld cordless magnetic probe following preoperative SLN mapping by MRL using SPIO is feasible, without RI use, for neck management in cases of clinically N0 early oral cancer.

Large dentigerous cyst associated with the maxillary impacted supernumerary teeth: A rare occurrence and literature review.

Dentigerous cysts are common odontogenic cysts of the jaw but are rarely associated with supernumerary teeth. Few cases of large dentigerous cysts associated with anterior maxillary supernumerary teeth have been reported. The English literature has documented only four cases of dentigerous cysts>40 mm in diameter associated with supernumerary teeth. A 47-year-old man was referred to our hospital, complaining of minor pain in the maxillary gingiva. Computed tomography revealed a well-defined oval unilocular radiolucent lesion (50×45×35 mm) in the right maxilla, including two impacted supernumerary teeth. A dentigerous cyst associated with impacted anterior maxillary supernumerary teeth was diagnosed. The two impacted teeth were surgically removed, and the cyst was enucleated using the Caldwell-Luc approach. Histopathology confirmed the diagnosis of a large dentigerous cyst associated with impacted anterior maxillary supernumerary teeth. The postoperative course has been uneventful for two years. We also reviewed the relevant English literature.

A prospective interventional trial on the effect of periodontal treatment on Fusobacterium nucleatum abundance in patients with colorectal tumours.

Fusobacterium nucleatum is associated with the progression of colorectal cancer. Thus, the possibility of preventing colorectal cancer or its progression by targeting F. nucleatum has been explored. As F. nucleatum is associated with periodontitis, we analysed whether treating periodontitis could influence F. nucleatum abundance in the colon. Patients with colorectal tumours who underwent colonoscopy were recruited. Patients diagnosed with periodontitis by a dentist were treated for approximately 3 months. Endoscopic resection of colorectal tumours was performed after periodontitis treatment, and resected tumours were pathologically classified as high-(HGD) or low-grade dysplasia (LGD). Saliva and stool samples were collected before and after the treatment. Of the 58 patients with colorectal tumours, 31 were included in the study, 16 showed improvement in periodontitis, and 11 showed no improvement. Stool F. nucleatum levels before treatment were significantly lower in the LGD group than in the HGD group. A significant decrease in faecal F. nucleatum levels was observed in patients who underwent successful treatment but not in those whose treatment failed. Salivary F. nucleatum levels were not altered in patients despite periodontal treatment. Thus, successful periodontitis treatment reduces stool F. nucleatum levels and may aid research on periodontitis and suppression of colorectal cancer development.

Transcriptomic Profiling Predicts Multiple Pathways and Molecules Associated With the Metastatic Phenotype of Oral Cancer Cells.

BACKGROUND/AIM: Metastasis to cervical lymph nodes of oral squamous cell carcinoma (OSCC) leads to a poor prognosis. The present study aimed at investigating the pathways and molecules associated with OSCC metastasis. MATERIALS AND METHODS: The transcriptome between HSC-3 cells and their highly metastatic subline, HSC-3-M3 cells, was examined using gene expression microarray. Gene enrichment analyses and Ingenuity Pathway Analysis were performed. Kaplan-Meier plot analysis using a publicly available dataset was conducted to assess whether candidate molecules are prognosticators. RESULTS: A total of 1,018 genes were differentially expressed, and the inflammatory pathway and NF-kB were predicted to be activated in HSC-3-M3 cells. CSF2 was suggested to be an indicator of poor prognosis in head and neck cancers. CONCLUSION: Inflammation and NF-kB may be involved in the metastasis of OSCC, and CSF2 is a promising diagnostic and therapeutic molecule. Moreover, HSC-3-M3 cells are a useful cell line model for studying OSCC progression.

Sentinel lymph node mapping of clinically N0 early oral cancer: a diagnostic pitfall on CT lymphography.

OBJECTIVE: The purpose of this study was to reevaluate preoperative computed tomography lymphography (CTL) and enhanced CT images during follow-up to clarify whether SLNs enhanced by CTL were identified accurately before primary surgery with sentinel lymph node biopsy (SLNB) for clinically N0 early oral cancer. METHODS: Thirty two early oral cancer patients without cervical lymph node metastasis were enrolled in this study. To clarify whether SLNs enhanced by CTL were identified accurately before primary surgery with SLNB, we reevaluated preoperative CTL and enhanced CT images during follow-up in all patients. RESULTS: SLNs were detected by CTL in 31 of 32 patients (96.9%). During follow-up after primary surgery with SLNB, 4 of 27 patients without SLN metastasis had occult neck metastasis. Of the 4 patients, only 1 patient with cancer of floor of the mouth had overlooking of SLN, and the overlooking rate of SLN was 3.1%. The overlooked small SLN (2.9 × 3.3 × 3.1 mm) was located at contralateral level IB. The CT numbers before, 2, 5, 10 min after iopamidol injection, were 33 HU, 37 HU, 62 HU, 52 HU, respectively. The CT numbers of overlooked SLN 5 and 10 min after the injection was higher than CT images scanned before the iopamidol injection. CONCLUSIONS: The enhancement of SLNs in CTL images after iopamidol injection should be compared sufficiently with CT images before iopamidol injection to avoid overlooking of SLNs in N0 early oral cancer.

MR lymphography with superparamagnetic iron oxide for sentinel lymph node mapping of N0 early oral cancer: A pilot study.

OBJECTIVES: The purpose of this pilot study was to evaluate the usefulness of magnetic resonance lymphography (MRL) with superparamagnetic iron oxide (SPIO) in sentinel lymph node (SLN) mapping of clinically N0 early oral cancer, and to conduct a comparative study of this MRL with CT lymphography (CTL). METHODS: CTL and MRL were performed for SLN mapping before surgery for 20 patients with clinically N0 early oral cancer. The detection rate, number, and location of SLNs in CTL and MRL were evaluated. Furthermore, optimal scanning/imaging timing in MRL was examined. RESULTS: SLNs were detected by CTL in 18 (90%) patients, and the total and mean number of SLN were 35 and 1.8, respectively. All SLNs could be detected 2 min and 3.5-5 min after contrast medium injection. In all patients, SLNs were detected by MRL at 10 min after SPIO injection, and the total and mean number of SLN was 53 and 2.7, respectively. MRL at 30 min after the injection showed additional 18 secondary lymph nodes. CONCLUSION: MRL with SPIO is safe and useful imaging for the detection of SLNs in clinically N0 early oral cancer, and the optimal imaging timing is 10 min after SPIO injection.

Sentinel lymph node biopsy with preoperative CT lymphography and intraoperative indocyanine green fluorescence imaging for N0 early tongue cancer: A long-term follow-up study.

PURPOSE: This study evaluated the usefulness of sentinel lymph node (SLN) biopsy with preoperative computed tomographic lymphography (CTL) and intraoperative indocyanine green (ICG) fluorescence imaging for N0 early tongue cancer. METHODS: Twenty-seven patients with N0 early oral tongue cancer underwent CTL with a 128-slice multi-detector row CT scanner to detect SLN on the day before resection of primary tumor and SLN biopsy under ICG fluorescence guidance. We identified the location and number of SLNs mapped by CTL and evaluated whether CTL-enhanced SLNs could be identified intraoperatively as ICG fluorescent lymph nodes. Prognosis was also evaluated. RESULTS: SLNs were detected by CTL in 26 of 27 patients (96.3%). The total and mean numbers of SLNs were 41 and 1.5, respectively. All SLNs enhanced by CTL could be identified intraoperatively as ICG fluorescent lymph nodes. Two SLNs were found under ICG fluorescent guidance in only one patient without SLN enhanced by CTL. Among the 27 patients, five (18.5%) had SLN with metastasis. Median follow-up was 76 months (range 44-82 months). During follow-up, three of 22 patients without SLN metastasis had occult cervical lymph node metastasis. The 5-year overall survival rate was 100%. CONCLUSION: SLN biopsy with preoperative CTL and intraoperative ICG fluorescence imaging is a feasible and reliable procedure, without radioisotope tracers, for neck management in cases of early tongue cancer.

CT lymphography for sentinel lymph node mapping of clinically N0 early oral cancer.

BACKGROUND: The objectives of this retrospective study were to evaluate the usefulness of computed tomography lymphography (CTL) and to clarify the optimal timing of CTL in sentinel lymph node (SLN) mapping of clinically N0 early oral cancer. METHODS: Twenty patients with clinically N0 early oral cancer underwent CTL with a 128 multi-detector row CT scanner to detect SLN the day before resection of primary tumor and SLN biopsy with indocyanine green (ICG) fluorescence guidance. CT scanning was performed in the first 10 patients at 2, 5, and 10 min after submucosal injection of iopamidol and in the remaining 10 patients at 2, 3.5, 5, and 10 min after the injection of contrast medium. We evaluated the SLN detection rate at each scan timing and the number and location of SLNs. We evaluated whether CTL-enhanced SLNs could be identified intraoperatively as ICG fluorescent lymph nodes. RESULTS: SLNs were detected by CTL in 19 of the 20 patients (95.0%), and the mean number of SLNs was 2 (range, 1-4). All SLNs were located on the ipsilateral side; 35 of 37 SLNs were located at level I and II, and 2 SLNs were lingual lymph nodes. All SLNs could be detected 2 min and 3.5-5 min after contrast medium injection, and CTL-enhanced SLNs could be identified intraoperatively as fluorescent lymph nodes. CONCLUSIONS: CTL could facilitate the detection of SLNs in early oral cancer, and the optimal timing of CT scanning was at 2 and 5 min after injection of contrast medium.

Atypical Protein Kinase C λ/ι Expression Is Associated with Malignancy of Oral Squamous Cell Carcinoma.

BACKGROUND/AIM: Atypical protein kinase C λ/ι (aPKCλ/ι) is a cell polarity-regulator localized in the tight junction and apical membrane in epithelial cells. Previous studies suggested that aPKCλ/ι overexpression and abnormal localization were involved in tumor progression in several cancers. We investigated the relationship between aPKCλ/ι and oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: The correlation between the aPKCλ/ι expression and the clinicopathological parameters in 76 OSCC cases was examined using immunohistochemical analyses. RESULTS: aPKCλ/ι overexpression was observed in 36.8% of cases. aPKCλ/ι expression was more intense in poorly differentiated OSCC and younger patients (<60 years of age). Although expression of aPKCλ/ι was not significantly associated with clinical parameters, the correlation was found between aPKCλ/ι localization and progression-free survival. CONCLUSION: This is the first study to assess the association of aPKCλ/ι expression in OSCC with clinical results. Expression and localization of aPKCλ/ι may be involved in the degree of malignancy in OSCC.

Multidisciplinary approach for treatment of a dentigerous cyst - marsupialization, orthodontic treatment, and implant placement: a case report.

BACKGROUND: Dentigerous cysts are common odontogenic cysts associated with unerupted teeth. We describe a previously unreported case of a multidisciplinary approach using surgical, orthodontic, and implant treatment to establish the occlusion for a patient with a maxillary dentigerous cyst. CASE PRESENTATION: An 18-year-old Japanese woman visited our hospital with a chief complaint of gingival swelling in her anterior maxillary region, midline diastema, and tooth crowding. Her main symptom was this gingival swelling. A panoramic radiograph revealed a radiolucent area, 30 mm in diameter, round in shape, and with well-demarcated margins including the maxillary canine. Computed tomography revealed a cystic cavity filled with homogeneous fluid of the same density as water, and a distolingually inclined canine. Our clinical diagnosis was maxillary dentigerous cyst with an unerupted distolingually inclined canine. The selected treatment was marsupialization of the dentigerous cyst, followed by orthodontic traction of the unerupted canine, and simultaneous orthodontic treatment of the midline diastema and tooth crowding. The orthodontic traction failed because the canine did not erupt completely, and the canine was extracted. The treatment plan was then changed to implant treatment after the tooth crowding and midline diastema had been improved. Because the alveolar ridge width was inadequate, the implant was placed after a two-stage implant treatment; therefore, a satisfactory occlusion could be achieved. Our patient did not experience any complications, and the cyst has not recurred. A radiograph taken 7 years after marsupialization of the dentigerous cyst revealed that the cystic cavity had been replaced by new bone. CONCLUSIONS: In general, orthodontic traction of an unerupted tooth after marsupialization should be the best option. However, if orthodontic traction fails, a multidisciplinary approach involving implant treatment may be necessary. We describe a case in which a multidisciplinary approach involving surgical, orthodontic, and implant treatment was used to establish a satisfactory occlusion for a patient with a dentigerous cyst.

Lateral Bone Window Approach with Micross Mini Bone Scraper for Sinus Floor Elevation.

OBJECTIVE: The purpose of this article is to describe a technique with a mini bone scraper for lateral bone window approach and simultaneous bone harvesting during sinus floor elevation. METHODS: This study included five consecutive patients with the atrophic posterior maxilla. After elevation of mucoperiosteal flap, a bone window was then opened safely using a Micross Mini Bone Scraper® with small shape and easy handling. Data were recorded on the residual alveolar bone height, number of implants, site of implant placement, collected particulate bone volume, and bone harvesting time. RESULTS: There was no injury of the Schneiderian membrane, and one to two implants of 11.5 mm length were placed simultaneously after sinus floor elevation in all cases. Mean residual bone height was 4.6 mm. The mean volume of particulate bone collected from the anterior wall of the maxilla using this technique was 0.74 cm3. Supplementary allogeneic materials were not required in all cases. Mean bone harvesting time was 4.6 min. CONCLUSIONS: Our technique with Micross Mini Bone Scraper® is a simple and safe procedure in lateral bone window approach and simultaneous bone harvesting during sinus floor elevation, because it is performed under better visualization of the membrane without irrigation.

Increase of peripheral blood CD57+ T-cells in patients with oral squamous cell carcinoma.

BACKGROUND/AIM: The subset of T-cells positive for expression of cluster of differentiation (CD) 57 has been associated with various cancer phenotypes. However, the presence of CD57(+) T-cells in patients with oral squamous cell carcinoma (OSCC) has yet to be confirmed. In the present study, we examined the diagnostic significance of the presence of CD57(+) T-cells in peripheral blood (PB) from patients with OSCC. MATERIALS AND METHODS: The subset of CD57(+) T-cells in PB was analyzed in 43 patients with OSCC by fluorescence-activated cell sorting (FACS) analysis. RESULTS: The proportion of CD57(+) T-cells, including both CD8(+) and CD4(+) subsets, significantly increased with clinical stage, especially in parallel with tumor size. CONCLUSION: Our results suggest that an increase in the population of CD57(+) T-cells is a potent prognostic marker and may also influence the systemic immunity of patients with OSCC.

Peripheral osteoma of the mandibular notch: report of a case.

Osteoma is a benign, slow-growing osteogenic tumor that sometimes arises from the craniomaxillofacial region, such as the sinus, temporal or jaw bones. Osteoma consists of compact or cancellous bone that may be peripheral, central or extraskeletal type. Peripheral osteoma arises from the periosteum and is commonly a unilateral, pedunculated mushroom-like mass. Peripheral osteoma of the mandible is relatively uncommon, and peripheral osteoma of the mandibular notch is extremely rare, although many cases arise from the mandibular body, angle, condyle, or coronoid process. We report here an unusual peripheral osteoma of the mandibular notch in a 78-year-old nonsyndromic female.

Warthin tumor arising from the minor salivary gland.

Warthin tumor (WT) accounts for 4% to 13% of all salivary gland tumors. This benign tumor, which commonly arises in the parotid gland, is the second most common tumor of the salivary gland. WT is multicentric in 12% to 20% of patients and is bilateral in 5% to 14%. The mean age at diagnosis is 62 years (range, 12-92 years), and it rarely presents (<6%) before age 40 years. Extraparotid WT, arising from the submandibular gland or cervical lymph node for example, is very infrequent, with corresponding incidences of 0.4% to 6.9% and 8%, respectively. Moreover, WT arising from the minor salivary gland is extremely rare, with a reported incidence of merely 0.1% to 1.2%. We report here WT arising from the minor salivary gland in the buccal mucosa in a 66-year-old woman and review cases of WT of the minor salivary gland reported in the English literature.

18F-fluorodeoxyglucose-positive Warthin tumor in a contralateral cervical lymph node mimicking metastasis in tongue cancer staging with PET/CT.

We report 18F-fluorodeoxyglucose-positive Warthin tumor in a contralateral cervical lymph node mimicking metastasis in tongue cancer staging with positron emission tomography/computed tomography.