To screen for additional treatment targets against tongue cancer, we evaluated the contributions of extracellular signal-related kinase (ERK), AKT and ezrin in cancer development. Immunohistochemical staining showed that ERK and ezrin expressions were significantly higher in invasive squamous cell carcinoma than in carcinoma in situ. To investigate the roles of ERK and ezrin in cancer development, we used the non-woven silica fibre sheet CellbedTM with a structure resembling the loose connective tissue morphology in a novel 3D culture system. We confirmed that the 3D system using CellbedTM accurately mimicked cancer cell morphology in vivo. Furthermore, cell projections were much more apparent in 3D-cultured tongue cancer cell lines than in 2D cultures. Typically, under conventional 2D culture conditions, F-actin and cortactin are colocalized in the form of puncta within cells. However, in the 3D-cultured cells, colocalization was mainly observed at the cell margins, including the projections. Projections containing F-actin and cortactin colocalization were predicted to be invadopodia. Although suppressing ezrin expression with small interfering RNA transfection caused no marked changes in morphology, cell projection formation was decreased, and the tumour thickness in vertical sections after 3D culture was markedly decreased after suppressing ERK activity because both the invasion ability and proliferation were inhibited. An association between cortactin activation as well as ERK activity and invadopodia formation was detected. Our novel 3D culture systems using Cellbed™ are simple and useful for in vitro studies before conducting animal experiments. ERK contributes to tongue cancer development by increasing both cancer cell proliferation and migration via cortactin activation.

Background Ezrin, ERK, STAT3, and AKT are proteins that are overexpressed in various types of cancer, although their expressions in tongue cancer has received less focus. This study aimed to address associations between the expression levels of these proteins and with characteristics of the tumor and patient survival. Methods We performed immunohistochemical staining of ezrin, ERK, STAT3, and AKT in tumors from patients with tongue carcinoma in situ (CIS, n = 17) and tongue squamous cell carcinoma (SCC, n = 46). Statistical differences between the SCC versus the CIS cohorts were estimated by calculations of bivariate odds ratios of low versus high expression of the proteins. Fisher's exact tests were used to appraise interassociations between the proteins, as well as expression levels versus patient and tumor characteristics. Survival based on Kaplan–Meier statistics in combination log‐rank tests were used to address potential effects of the patient and tumor characteristics versus 5‐year survival rate. Results The relative high: low expression of all four proteins in the two cohorts differed, and particularly ERK was markedly overexpressed in the SCC versus the CIS cohort (odds ratio = 45.3, p < .01). The relative high: low expression each protein versus patient and tumor characteristics; showed associations between AKT expression and T stage (p = .002) plus node metastases (p = .12), and between ERK expression and drinking (p = .01) and smoking history (p = .01). There was no significant difference observed between ERK and the three other molecules, nor any significant difference between the degree of expression of each protein and the 5‐year disease‐specific survival rate. Conclusion Ezrin, ERK, STAT3, and AKT appear to be involved in the progress from carcinoma in situ in the tongue into squamous cell carcinoma. ERK in particular is overexpressed, suggesting that ERK may be a novel therapeutic target for preventing tongue cancer.

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Background/AimThe first objective of this study was to identify predictive factors for oral and maxillofacial fractures at the initial response to the patient. The second objective was to determine the factors influencing the incidence of treatment duration of more than 1 month using the information shown in the medical record.Materials and MethodsHospital records from 2011 to 2019 were reviewed to identify patients who had sustained oral and maxillofacial injuries by falling or falling from a height. Patterns and types of oral and maxillofacial injury, injury severity, and background of the injury were collected from the hospital records. The variables independently associated with a treatment duration of more than 1 month were determined by logistic regression analysis.ResultsIn total, 282 patients (150 men, 132 women; median age, 17.5 years) were selected for analysis. Maxillofacial fractures were observed in 20.9% of patients (59/282); among these, mandibular fractures were the most common (47/59). Logistic regression analysis showed that age (odds ratio [OR], 1.026), nighttime occurrence (OR, 2.192), and upper face injury (OR, 20.704) were independent predictive factors for having a maxillofacial fracture. Additionally, the number of injured teeth (OR, 1.515) and the use of intermaxillary fixation (OR, 16.091) were independent predictors of treatment duration of more than 1 month.ConclusionsThese results may be useful in the initial management of maxillofacial injuries in terms of better‐informing patients injured by falling their expected treatment duration and managing the psychological impacts of a long treatment duration.

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