Tongue squamous cell carcinoma (TSCC) occurs frequently in the oral cavity, and because of its high proliferative and metastatic potential, it is necessary to develop a novel treatment for it. We have reported the importance of the inhibition of the periostin (POSTN) pathological splicing variant, including exon 21 (PN1-2), in various malignancies, but its influence is unclear in tongue cancer. In this study, we investigated the potential of POSTN exon 21-specific neutralizing antibody (PN21-Ab) as a novel treatment for TSCC. Human PN2 was transfected into the human TSCC (HSC-3) and cultured under stress, and PN2 was found to increase cell viability. PN2 induced chemotherapy resistance in HSC-3 via the phosphorylation of the cell survival signal Akt. In tissues from human TSCC and primary tumors of an HSC-3 xenograft model, PN1-2 was expressed in the tumor stroma, mainly from fibroblasts. The intensity of PN1-2 mRNA expression was positively correlated with malignancy. In the HSC-3 xenograft model, CDDP and PN21-Ab promoted CDPP\'s inhibition of tumor growth. These results suggest that POSTN exon 21 may be a biomarker for tongue cancer and that PN21-Ab may be a novel treatment for chemotherapy-resistant tongue cancer. The treatment points towards important innovations for TSCC, but many more studies are needed to extrapolate the results.

OBJECTIVE: To study and analyze the clinical patterns of lower-level lymph node (Ⅳ and Ⅴ) metastasis in primary tongue squamous cell carcinoma, and establish a reference for the decision-making of the lower-level neck dissection in tongue squamous cell carcinoma.
METHODS: A total of 203 patients with primary tongue squamous cell carcinoma were recruited. These patients underwent simultaneous/secondary comprehensive neck (level Ⅰ-Ⅴ) dissection in Shanghai Ninth People\'s Hospital, Shanghai Jiao Tong University School of Medicine from January 2010 to December 2015. Their clinicopathological and follow-up data were obtained and analyzed to reveal the prognosis and risk factors of primary tongue squamous cell carcinoma with lower-level lymph node metastasis.
RESULTS: Among the 203 patients, the metastasis rates of levels Ⅳ and Ⅴ are 14.78% and 4.93%, respectively. Level Ⅳ metastasis is more prevalent in males than females (P=0.04); non-smokers are more likely to have level Ⅴ metastasis than smokers (P=0.046). Lymph node status in levels Ⅲ and Ⅳ are significantly associated with the risk of metastasis in level Ⅴ (P=0.001). Patients with extracapsular invasion in level Ⅲ have a significantly increased risk of metastasis in levels Ⅳ (P=0.014) and Ⅴ (P=0.026). The 5-year survival rate of patients with lower lymph node (Ⅳ/Ⅴ) metastasis is only 14.70%, which is an independent poor prognostic factor for patients with primary tongue squamous cell carcinoma (P<0.000 1).
CONCLUSIONS: Most primary tongue squamous cell carcinoma metastases occur in levels Ⅰ-Ⅲ. However, the rate of lower-level lymph node metastasis is rather low. For the cN0 and cN+ patients with levels Ⅰ-Ⅱ lymph node metastases without extracapsular invasion, the strategy for level Ⅴ management may be observation rather than dissection.
目的: 研究分析原发性舌鳞状细胞癌颈部淋巴结低位（Ⅳ区和Ⅴ区）转移的临床规律，为舌癌颈部淋巴结低位区域的清扫决策提供参考依据。方法: 选取2010年1月—2015年12月上海交通大学医学院附属第九人民医院收治的原发性舌鳞状细胞癌同期或二期行全颈（Ⅰ~Ⅴ区）淋巴结清扫患者203例，收集其临床病理资料及随访信息，分析原发舌鳞状细胞癌颈部低位淋巴结转移的临床病理影响因素和预后。结果: 在203例患者中，Ⅳ区和Ⅴ区转移率分别为14.78%和4.93%，男性较女性更易出现Ⅳ区转移（P=0.04），不吸烟较吸烟更易出现V区转移（P=0.046）；Ⅲ区和Ⅳ区的淋巴结状态与Ⅴ区转移风险明显相关（P=0.001）；Ⅲ区包膜外侵犯患者发生Ⅳ区（P=0.014）和V区（P=0.026）转移的风险明显增加；发生低位淋巴结（Ⅳ/Ⅴ）转移的患者5年生存率仅为14.70%，是原发性舌鳞状细胞癌患者的独立不良预后因素（P<0.000 1）。结论: 原发性舌鳞状细胞癌绝大部分转移发生于Ⅰ~Ⅲ区，低位淋巴结转移比率较低，对于cN0患者和Ⅰ~Ⅱ区淋巴结转移但无包膜外侵犯的cN+患者，Ⅴ区淋巴结可以选择观察。.

BACKGROUND: IgG4, which plays a pivotal role in the progression of phenotypically diverse tumors, serves as a prognostic marker because of its influence on cancer immunity. Nevertheless, the functions of IgG4 in tongue squamous cell carcinoma (TSCC) remained to be identified.
METHODS: To evaluate the significance of IgG4 expression in TSCC, we performed immunohistochemical analysis of patients with TSCC (n = 50) to evaluate the correlation of IgG4 expression with patients\' clinicopathological features and prognoses.
RESULTS: Higher IgG4 expression detected in TSCC tissues was associated with the less advanced mode of invasion (Yamamoto-Kohama [YK] 1-3) (P = 0.031) and with well-differentiated TSCC (P = 0.077). Kaplan-Meier analyses revealed that the higher IgG4 expression group exhibited better prognosis indicated by overall survival (OS) (P = 0.04) and recurrence-free survival (RFS) (P = 0.016). Univariate analysis of OS indicated that IgG4 expression was associated with longer OS (P = 0.061), and multivariate analysis of RFS revealed that IgG4 expression served as an independent prognostic factor for longer RFS (P = 0.005).
CONCLUSIONS: These results indicate that relatively higher IgG4 levels serve as a favorable prognostic factor for TSCC.

Surgery is the primary treatment for tongue cancer. The goal is a complete resection of the tumor with an adequate margin of healthy tissue around the tumor.Inadequate margins lead to a high risk of local cancer recurrence and the need for adjuvant therapies. Ex vivo imaging of the resected surgical specimen has been suggested for margin assessment and improved surgical results. Therefore, we have developed a novel three-dimensional (3D) ultrasound imaging technique to improve the assessment of resection margins during surgery. In this research protocol, we describe a study comparing the accuracy of 3D ultrasound, magnetic resonance imaging (MRI), and clinical examination of the surgical specimen to assess the resection margins during cancer surgery. Tumor segmentation and margin measurement will be performed using 3D ultrasound and MRI of the ex vivo specimen. We will determine the accuracy of each method by comparing the margin measurements and the proportion of correctly classified margins (positive, close, and free) obtained by each technique with respect to the gold standard histopathology.

OBJECTIVE: Grape seed oil (GSO) has recently gained popularity due to its anticancer properties. This study aimed to investigate the efficacy of combining cisplatin (CP) and GSO in tongue squamous cell carcinoma (TSCC) treatment.
METHODS: In this study, human tongue carcinoma cell line (HNO-97) was treated with CP and GSO alone or in combination. The effects of CP and GSO on cytotoxicity and cell cycle arrest were studied using the MTT assay and flowcytometry, respectively. The apoptotic markers, including p53 and caspase 8, were assessed using reverse-transcription polymerase chain reaction (RT-PCR), caspase 3 using immunohistochemistry, and the angiogenic marker vascular endothelial growth factor (VEGF) using enzyme-linked immunosorbent assay (ELISA).
RESULTS: The IC50 drug concentrations were found to be 16.4 ug/mL of GSO and 2.18 ug/mL of CP. When compared to the untreated control group, the percentage of S phase cells and apoptotic cells was significantly higher in the GSO, CP, and GSO/CP combination therapy groups. Furthermore, p53, caspase 8, caspase 3 expression were significantly upregulated in the GSO-and CP-treated groups, with evident upregulation with GSO/CP combination therapy. However, VEGF levels were significantly lower in the GSO-, CP-, and combined GSO/CP-treated groups.
CONCLUSIONS: GSO has both an apoptotic and antiangiogenic effect in the treatment of TSCC, suggesting a new strategy for phytochemical-based combination therapy.

OBJECTIVE: In order to predict the patients\' prognosis with tongue squamous cell carcinoma (SCC), this study set out to develop a clinically useful and trustworthy prognostic nomogram.
METHODS: The Surveillance, Epidemiology, and End Results (SEER) Program was used to compile clinical information on patients with tongue SCC between 2010 and 2015. The likelihood of Cancer-Specific Survival (CSS) and Overall Survival (OS) for specific patients was predicted using a prognostic nomogram created with the help of the RStudio software. The nomogram\'s predictive ability was evaluated using the consistency index (C-index) and decision curve analysis, and the nomogram was calibrated for 1-, 2-, 3-, 5-, and 10-year CSS and OS.
RESULTS: Patients numbering 6453were enrolled in this study. The primary cohort (3895) and validation cohort (2558) were each randomly assigned. Sex, age, tumor-node-metastasis (TNM) stage, surgery, chemotherapy, and radiation were significant risk factors for OS, whereas age, TNM stage, surgery, chemotherapy, and radiotherapy were significant risk factors for CSS. Additionally, C-index and calibration curves indicated that the prognostic nomogram prediction and the actual observation in both cohorts would be very coherent.
CONCLUSIONS: The predictive nomogram created in this study can offer patients with tongue SCC customized treatment and survival risk assessment.

BACKGROUND: This study aimed to evaluate the prognostic outcome for adolescent and young adult (AYA) generation patients with tongue squamous cell carcinoma (TSCC).
METHODS: Data were obtained from the Head and Neck Cancer Registry of Japan, and patients who were newly diagnosed with TSCC from 2011 to 2014 were extracted. We compared the clinical parameter and survival of the ≤ 39 years old (AYA) patient group with the 40-79 (non-AYA) group. Propensity score matching (PSM) was used for survival analyses.
RESULTS: Our cohort included 2221 patients with TSCC. AYA and non-AYA groups consisted of 258 and 1963 patients, respectively. The AYA group has a larger proportion of females than the non-AYA group (P < 0.001). Following PSM, both overall and disease-specific survival of the AYA group was significantly longer than those of the non-AYA group (P = 0.009 and P = 0.04, respectively).
CONCLUSIONS: We demonstrated the survival superiority of AYA patients with TSCC compared to older adult patients. Therefore, our study results may reduce this anxiety by providing patients with appropriate information of prognosis for AYA patients with TSCC.

High-definition ultrasonography is a diagnostic tool that uses sound echoes to produce images of tissues and organs. In the head and neck region, ultrasounds have been used to diagnose different types of lesions. The intraoral approach was shown to be a real-time, non-invasive way to characterize oral lesions. The tongue is the most often examined region because of its accessibility. This observational study aimed to describe the qualitative characteristics of tongue squamous cell carcinoma images obtained with high-definition intraoral ultrasound by comparing them with the corresponding histopathological sample. Twenty patients were enrolled in this study. The scans of the lesions were carried out with an 18 MHz linear ultrasound probe following the long axis of the lesion. For each lesion, five frames were selected, on which descriptive analysis was performed. A histological sample was taken and then compared to the ultrasonographic acquisition. The sonographic appearance of the tissue layers has a good correlation between ultrasound and histological morphology, and it was easy to distinguish the tumor from the homogenous composition of the tongue tissues. Furthermore, a correlation between the structure by section and pattern of tumor margin features by ultrasound was obtained. Intraoral ultrasonography appears to be a promising technique in the non-invasive characterization of tongue squamous cell carcinoma. Further studies will be needed to improve the technique in terms of ergonomics and repeatability.

OBJECTIVE: The present in vitro study aims to investigate the potential use of epigenetic inhibitors as treatment modalities in tongue squamous cell carcinoma.
METHODS: The human tongue squamous cell carcinoma cell line (CAL-27) was cultured and exposed to varying concentrations of 5-Azacitidine (5-Aza) or Trichostatin A (TSA) in the culture medium. The cell apoptosis was evaluated using Annexin V/PI by flow cytometry. To evaluate DNA damage response, γH2AX foci analysis was performed using immunofluorescence. Single cell gel electrophoresis (SCGE) was applied to measure DNA strand breaks. Gene expression was assessed by quantitative real-time PCR.
RESULTS: The results showed that 5-Aza and TSA had apoptotic effects on the SCC cell line at concentrations of 50-200 µM and 0.5-5 µM, respectively. Immunofluorescence analysis showed increased expression of γH2AX, the marker of DNA damage response after treatment of 5-Aza and TSA that was associated with increased DNA strand breaks. The expressions of urokinase plasminogen activator, its receptor and matrix metalloproteinase-2, were significantly reduced in TSA- and 5-Aza-treated cells.
CONCLUSIONS: Our results showed that 5-Aza and TSA increase apoptotic and DNA damage response in squamous cell carcinoma cell line while reducing the expression of tumor invasion genes that further indicating the potential therapeutic value of two epigenetic modifiers in squamous cell carcinoma.

Compared with traditional imaging techniques, multimodal imaging obtains more accurate images that may increase disease detection rates. The present study prepared stromal cell-derived factor 1 (SDF-1)-loaded, targeted nanoparticles coated with iron (II,III) oxide and perfluorohexane (PFH) to be used as polymer-shelled contrast agents with multimodal imaging functions, with the aim of improving tongue cancer and lymph node metastasis diagnosis. The multifunctional, targeted, polymeric nanoparticles were prepared using a double emulsion method and chemokine SDF-1 was conjugated to nanoparticles by a sulfide bond. The nanoparticles were spherical, uniform size and well dispersed. The results of the in vitro photoacoustic and ultrasonic imaging experiments demonstrated that the multifunctional nanoparticles displayed excellent multimodal imaging functions, as even small concentrations of nanoparticles presented clear ultrasound and photoacoustic imaging. When the temperature reached the boiling point of PFH (56˚C), a liquid-gas phase change occurred and the microsphere volume and acoustic impedance increased, leading to enhanced ultrasonic development. The nanoparticles were automatically targeted to tongue squamous carcinoma cells in vitro via SDF-1-CXC chemokine receptor 4 interactions. The targeted experiment and flow cytometry results indicated that the nanoparticles underwent strong targeted binding to human tongue squamous cell carcinoma (SCC-15) cells. In summary, the nanoparticles were automatically targeted to SCC-15 cells and displayed promising characteristics for ultrasound and photoacoustic imaging. Higher concentrations of nanoparticles was associated with clearer imaged and greater echo intensity value and photoacoustic value. The present study established a foundation for the development of procedures for primary tongue cancer and lymph node metastasis diagnosis.