To identify genetic alterations associated with tongue cancer recurrence in young adults, whole exome sequencing of the primary tumor, recurrence, and whole blood samples from young patients with tongue cancer was performed. A frameshift mutation in the TP53 gene was detected in the primary tumor and recurrence tumor tissue. A mutation in the EPHB6 gene was detected in the recurrence and was absent in the primary tumor. In addition, the primary tumor and recurrence tongue cancer tissue harbored amplification of the 20p13 region containing C20orf96, DEFB125, DEFB126, DEFB127, DEFB128, DEFB129, DEFB132, and ZCCHC3 genes. Thus, genetic alterations have been identified that are associated with tongue cancer recurrence in young adults.

OBJECTIVE: Radiomics is an emerging field that utilizes quantitative features extracted from medical images to predict clinically meaningful outcomes. Validating findings is crucial to assess radiomics applicability. We aimed to validate previously published magnetic resonance imaging (MRI) radiomics models to predict oncological outcomes in oral tongue squamous cell carcinoma (OTSCC).
METHODS: Retrospective multicentric study on OTSCC surgically treated from 2010 to 2019. All patients performed preoperative MRI, including contrast-enhanced T1-weighted (CE-T1), diffusion-weighted sequences and apparent diffusion coefficient map. We evaluated overall survival (OS), locoregional recurrence-free survival (LRRFS), cause-specific mortality (CSM). We elaborated different models based on clinical and radiomic data. C-indexes assessed the prediction accuracy of the models.
RESULTS: We collected 112 consecutive independent patients from three Italian Institutions to validate the previously published MRI radiomic models based on 79 different patients. The C-indexes for the hybrid clinical-radiomic models in the validation cohort were lower than those in the training cohort but remained > 0.5 in most cases. CE-T1 sequence provided the best fit to the models: the C-indexes obtained were 0.61, 0.59, 0.64 (pretreatment model) and 0.65, 0.69, 0.70 (posttreatment model) for OS, LRRFS and CSM, respectively.
CONCLUSIONS: Our clinical-radiomic models retain a potential to predict OS, LRRFS and CSM in heterogeneous cohorts across different centers. These findings encourage further research, aimed at overcoming current limitations, due to the variability of imaging acquisition, processing and tumor volume delineation.

An artificial intelligence (AI) model was designed to assist pathologists in diagnosing and quantifying structural changes in tongue lesions induced by chemical carcinogens. Using a tongue cancer model induced by 4-nitroquinoline-N-oxide and treated with β-elemene, a total of 183 digital pathology slides were processed. The Segment Anything Model (SAM) was employed for initial segmentation, followed by conventional algorithms for more detailed segmentation. The epithelial contour area was computed using OpenCV\'s findcontour method, and the skeletonize method was used to calculate the distance map and skeletonized representation. The AI model demonstrated high accuracy in measuring tongue epithelial thickness and the number of papilla-like protrusions. Results indicated that the model group had significantly higher epithelial thickness and fewer papillae compared with the blank group. Furthermore, the treatment group exhibited reduced epithelial thickness and fewer papilla-like protrusions compared with the model group, though these differences were less pronounced. Overall, the SAM framework algorithm proved effective in quantifying tongue epithelial thickness and the number of papilla-like protrusions, thereby assisting healthcare professionals in understanding pathological changes and assessing treatment outcomes.

OBJECTIVE: Anterior 2/3rds glossectomy results in significant patient morbidity due to speech and swallowing impairment. Microvascular free flap reconstruction compensates for large volume defects. Flap volume is based on the adipose content of the donor site and varies by patient body mass index (BMI) and donor site location. We sought to correlate flap thickness at different donor sites with patient BMI to determine optimal donor site selection.
METHODS: Patients with CT scans of the oral cavity, thorax and lower extremity were identified and included. The volumes of the anterior 2/3rds of the tongue were measured and recorded using computed tomography-generated modeling. Pre-muscular tissue thicknesses at anterolateral thigh (ALT), deep inferior epigastric artery (DIEP), latissimus dorsi, and parascapular donor sites were measured. The donor site adequency was defined as reconstructing the tongue volume within 10% of the ideal volume required and stratified based on patient BMI.
RESULTS: In 144 patients, the average anterior 2/3rds glossectomy defect was 100.3 cm3. Glossectomy defect size was highly correlated with BMI (p < 0.001). The DIEP flap had the largest volume (155.4 cm3), followed by latissimus (105.6 cm3), parascapula (97.8 cm3), and ALT (60.5 cm3). For patients with BMI ≤ 30, the DIEP flap best reconstructed native tongue volume (up to 113 % of native tongue volume). In patients with BMI > 30.1, native tongue volumes were approximated by the latissimus flap (89-92 % of native tongue) and parascapular flap (85-95 % of native tongue volume). In BMI > 30.1 the DIEP flap provided excess tissue bulk (129-135 % of native tongue volume).
CONCLUSIONS: The DIEP flap more closely approximates the volume needed to reconstruct anterior two-thirds tongue defects for BMIs ≤ 30. The subscapular system flaps provided the best volume match for BMIs > 30 and the DIEP flap provided excess tissue bulk which could be adjusted in the reconstruction process.

OBJECTIVE: This study aims to assess the role of DNA methylation changes in tongue cancer through a comprehensive analysis of global DNA methylation alterations during experimental lingual carcinogenesis.
METHODS: C57BL/6J mice were subjected to 16-week oral administration of 4-nitroquinoline-1-oxide (4NQO, 50 mg/L). Lingual mucosa samples, being representative of normal tissue (week 0) and early (week 12) and advanced (week 28) tumorigenesis, were harvested for microarray and methylated DNA immunoprecipitation sequencing (MeDIP-Seq). The mRNA and promoter methylation of transforming growth factor-beta-signaling protein 1 (SMAD1) were evaluated with real-time quantitative reverse transcription polymerase chain reaction and Massarray in human lingual mucosa and tongue cancer cell lines.
RESULTS: The cytosine guanine island (CGI) methylation level observed at 28 weeks surpassed that of both 12 weeks and 0 weeks. The promoter methylation level at 12 weeks exceeded that at 0 weeks. Notably, 208 differentially expressed genes were negatively correlated to differential methylation in promoters among 0, 12, and 28 weeks. The mRNA of SMAD1 was upregulated, concurrent with a decrease in promoter methylation levels in cell lines compared to normal mucosa.
CONCLUSIONS: DNA methylation changed during lingual carcinogenesis. Overexpression of SMAD1 was correlated to promoter hypomethylation in tongue cancer cell lines.
目的: 研究舌黏膜癌变过程中基因组甲基化特征，探讨舌癌中DNA甲基化的规律。方法: 用50 mg/L的4-硝基喹啉-1-氧化物（4NQO）饮水诱导C57BL/6J小鼠舌黏膜癌变，分别取第0、12、28周的舌黏膜（分别代表正常、癌前病变和癌变）进行基因芯片检测和甲基化DNA免疫沉淀测序（MeDIP-Seq），在人舌黏膜组织和人舌癌细胞系中，用实时定量逆转录聚合酶链反应（qRT-PCR）和飞行质谱检测验证转化生长因子贝塔信号蛋白1（SMAD1）的表达和启动子的甲基化。结果: 28周较12周和0周舌黏膜的胞嘧啶鸟嘌呤岛（CGI）甲基化水平均升高，12周时启动子甲基化水平高于0周。在0、12和28周期间，208个差异表达基因与启动子中的差异甲基化呈负相关。与正常黏膜相比，细胞系中SMAD1的mRNA上调，同时启动子甲基化水平降低。结论: 舌黏膜癌变中伴随DNA甲基化修饰异常，舌癌中SMAD1高表达伴启动子低甲基化。.

BACKGROUND: Robotic neck dissection is emerging as an alternative to conventional open neck dissection. However, the oncologic safety of robotic elective neck dissection (END) and its indications in early-stage tongue cancer are unclear.
METHODS: We retrospectively reviewed the data of 78 patients who underwent transoral excision for T1, T2 squamous cell carcinoma of tongue with simultaneous ipsilateral END. Patients were assigned to two groups: the robotic group (n = 32)-postauricular face-lift -and the conventional group (n = 46)- transcervical incision. We compared the survival, clinical, pathologic and cosmetic outcomes of the two groups, and evaluated the number of retrieved lymph nodes and robot console time in the robotic group.
RESULTS: The mean age was lower in the robotic group (43.6 ± 12.8 vs. 55.8 ± 14.0, p < 0.001) and the conventional group had more T2 patients (p = 0.01). The mean operation time was significantly longer in the robotic group than the conventional group (178.81 ± 33.9 vs. 92.28 ± 16.7, p < 0.001). The mean number of retrieved lymph nodes was not significantly different between the two groups (19.22 ± 8.51 vs. 20.7 ± 11.4, p = 0.41). The 5-year disease-free survival rate was not significantly different between the two groups (93.6 % vs. 82.9 %, p = 0.59). Overall scar satisfaction assessed by VAS score, the robotic group showed significantly better results compared to the conventional group (8.38 vs. 5.86, p = 0.033).
CONCLUSIONS: Robotic END by a postauricular facelift approach is a feasible and safe approach for early-stage tongue cancer.

This study proposes a scoring system for adjuvant irradiation for stage I/II oral squamous cell carcinoma (OSCC). Derivation cohort (119 patients, operated between 2011 and 2014) and a validation cohort (204 patients, operated between 2016 and 2019) were included. In derivation cohort, on univariate analysis, tumor size >2 cm [3-year Disease Free Survival (DFS) 72.5% vs 95.6%, P = 0.039], lymphovascular invasion (58.3% vs 83.6%, P = 0.024), perineural invasion (75% vs 85.6%, P = 0.013), and depth of invasion ≥0.5 cm (73.8% vs 97.5%, P = 0.017) predicted 3-year DFS. Tongue lesions and poor differentiation were added as poor prognosticators based on previously published reports. Patients were grouped as low risk (<3 risk factors) and high risk (≥3 risk factors), with only high-risk group receiving adjuvant irradiation in validation cohort. Overall, 47/119 (39.5%) patients in the derivation cohort and 50/204 (24.5%) patients in validation cohort received adjuvant irradiation. In derivation cohort, 3-year DFS was 93% and 72.5% in the low and high-risk group, respectively. 3-year DFS was 90.7% and 85.8% in the low and high-risk group, respectively for validation cohort. The proposed scoring system reduced the use of adjuvant irradiation by 38%, with similar DFS.

BACKGROUND: In the treatment of oral cavity cancer, margin status is one of the most critical prognostic factors. Positive margins are associated with higher local recurrence and lower survival rates. Therefore, the universal goal of oral surgical oncology is to achieve microscopically clear margins. Near-infrared fluorescence guided surgery (FGS) could improve surgical resection using fluorescent probes. αVβ6 integrin has shown great potential for cancer targeting due to its overexpression in oral cancers. Red fluorescent contrast agent IRDye 680 coupled with anti-αVβ6 peptide (IRDye-A20) represents an asset to improve FGS of oral cancer. This study investigates the potential of IRDye-A20 as a selective imaging agent in 3D three-dimensional tongue cancer cells.
METHODS: αVβ6 integrin expression was evaluated by RT-qPCR and Western Blotting in 2D HSC-3 human tongue cancer cells and MRC-5 human fibroblasts. Targeting ability of IRDye-A20 was studied in both cell lines by flow cytometry technique. 3D tumor spheroid models, homotypic (HSC-3) and stroma-enriched heterotypic (HSC-3/MRC-5) spheroids were produced by liquid overlay procedure and further characterized using (immuno)histological and fluorescence-based techniques. IRDye-A20 selectivity was evaluated in each type of spheroids and each cell population.
RESULTS: αVβ6 integrin was overexpressed in 2D HSC-3 cancer cells but not in MRC-5 fibroblasts and consistently, only HSC-3 were labelled with IRDye-A20. Round shaped spheroids with an average diameter of 400 μm were produced with a final ratio of 55%/45% between HSC-3 and MRC-5 cells, respectively. Immunofluorescence experiments demonstrated an uniform expression of αVβ6 integrin in homotypic spheroid, while its expression was restricted to cancer cells only in heterotypic spheroid. In stroma-enriched 3D model, Cytokeratin 19 and E-cadherin were expressed only by cancer cells while vimentin and fibronectin were expressed by fibroblasts. Using flow cytometry, we demonstrated that IRDye-A20 labeled the whole homotypic spheroid, while in the heterotypic model all cancer cells were highly fluorescent, with a negligible fluorescence in fibroblasts.
CONCLUSIONS: The present study demonstrated an efficient selective targeting of A20FMDV2-conjugated IRDye 680 in 3D tongue cancer cells stroma-enriched spheroids. Thus, IRDye-A20 could be a promising candidate for the future development of the fluorescence-guided surgery of oral cancers.

UNASSIGNED: Adenoid cystic carcinoma (ACC) is an uncommon malignancy of head and neck. Although the cornerstone of treatment is surgery, concurrent chemoradiotherapy (CRT) might be used as an effective treatment for unresectable tumors. Herein we report a case of massive ACC of base of tongue with durable complete response to definitive CRT.
UNASSIGNED: Adenoid cystic carcinoma (ACC) is a rare tumor accounting for 1% of all head and neck cancers. The best treatment option is complete surgical resection with or without adjuvant radiotherapy. When surgical resection is not feasible, definitive radiotherapy with or without concurrent chemotherapy can be considered. Herein we report a non-smoker 72-year-old woman presented with throat discomfort and sensation of a lump. Evaluation revealed an unresectable adenoid cystic carcinoma of the base of tongue in whom complete clinical response was achieved after definitive concurrent chemoradiation. Although the cornerstone of treatment is complete surgical resection, this case report indicates that concurrent chemoradiotherapy might result in complete clinical response and could be used as a definitive treatment in selected ACC tumors.

UNASSIGNED: To avoid the oncologic risks of ipsilateral regional flaps, this study aimed to explore the feasibility and clinical outcomes of the contralateral-based facial artery myomucosal island flap (C-FAMMIF) for oral T2-T3 oncologic defects reconstruction.
UNASSIGNED: A study of flap anatomy was conducted on 7 cadaver samples and a cohort of 24 patients who received C-FAMMIF reconstruction after malignancy resection were retrospectively researched. A balanced anterolateral thigh flap (ALT) group of 47 patients was extracted as control group using propensity score matching method. Progression-free survival (PFS), functional outcomes, and donor site complications were assessed.
UNASSIGNED: Consistent blood supply and drainage through facial artery and vein with median maximum pedicle length of 106 mm supported contralateral reconstruction. The superficial vein drainage pattern indicated safer flap harvest at contralateral neck under circumstances of ipsilateral neck dissections. The pedicle and marginal facial nerve formed three anatomical patterns. The surgical management of each was described. Patients with ipsilateral pN+ neck accounted for 41.7% and 40.4% in the C-FAMMIF and ALT group, respectively. The 2-year PFS rate between the C-FAMMIF and ALT groups was not significantly different (88.2% in C-FAMMIF group and 84.6% in ALT group, respectively, p = 0.6358). Promising recoveries were observed for swallowing function and tactile sensation. The donor sites healed upon primary closure without trismus or permanent facial palsy.
UNASSIGNED: Our findings suggested that C-FAMMIF is feasible and safe for T2-T3 oral oncologic defect reconstruction in patients with ipsilateral cN+ neck.