Pannexin 1-Mediated ATP Signaling in the Trigeminal Spinal Subnucleus Caudalis Is Involved in Tongue Cancer Pain.

Pain is one of the most severe concerns in tongue cancer patients. However, the underlying mechanisms of tongue cancer pain are not fully understood. We investigated the molecular mechanisms of tongue cancer-induced mechanical allodynia in the tongue by squamous cell carcinoma (SCC) inoculation in rats. The head-withdrawal threshold of mechanical stimulation (MHWT) to the tongue was reduced following SCC inoculation, which was inhibited by intracisternal administration of 10Panx, an inhibitory peptide for pannexin 1 (PANX1) channels. Immunohistochemical analyses revealed that the expression of PANX1 was upregulated in the trigeminal spinal subnucleus caudalis (Vc) following SCC inoculation. The majority of PANX1 immunofluorescence was merged with ionized calcium-binding adapter molecule 1 (Iba1) fluorescence and a part of it was merged with glial fibrillary acidic protein (GFAP) fluorescence. Spike frequencies of Vc nociceptive neurons to noxious mechanical stimulation were significantly enhanced in SCC-inoculated rats, which was suppressed by intracisternal 10Panx administration. Phosphorylated extracellular signal-regulated kinase (pERK)-immunoreactive (IR) neurons increased significantly in the Vc after SCC inoculation, which was inhibited by intracisternal 10Panx administration. SCC inoculation-induced MHWT reduction and increased pERK-IR Vc neuron numbers were inhibited by P2X7 purinoceptor (P2X7R) antagonism. Conversely, these effects were observed in the presence of P2X7R agonist in SCC-inoculated rats with PANX1 inhibition. SCC inoculation-induced MHWT reduction was significantly recovered by intracisternal interleukin-1 receptor antagonist administration. These observations suggest that SCC inoculation causes PANX1 upregulation in Vc microglia and adenosine triphosphate released through PANX1 sensitizes nociceptive neurons in the Vc, resulting in tongue cancer pain.

Model of Human Tongue Squamous Cell Lines Stably Transfected with Human Papillomavirus (HPV)16 E6 and E7 Genes and Biological Characteristic Analysis.

BACKGROUND: Human tongue squamous cell carcinoma (TSCC) is the most common oral cancer with the highest human papillomavirus (HPV) infection rate in oral cancer. The purpose of this study was to research the correlation between HPV and TSCC. METHOD: Plasmid pEGFP/HPV16 E6E7 and plasmid pEGFP/no HPV16 E6E7 were constructed. TSCC cell lines SCC9 and SCC15 were infected by liposome transfection and would be highly selected by antibiotic. Fluorescence imaging, PCR, and Western blot were used to detect the expression of HPV16 E6E7 in cells. The biological characteristics were detected by CCK-8, wound healing assay, qRT-PCR, and Western blot. RESULT: TSCC cell lines transfected with HPV16 E6E7 gene were successfully established and identified. And the proliferation and migration ability of the TSCC cell lines infected with HPV16 E6E7 gene were significantly stronger than that of the blank group. CONCLUSION: TSCC cell lines infected with HPV16 E6E7 with significantly higher ability of proliferation and migration were more malignant than those not infected with HPV16 E6E7.

Data mining of an acoustic biomarker in tongue cancers and its clinical validation.

The promise of speech disorders as biomarkers in clinical examination has been identified in a broad spectrum of neurodegenerative diseases. However, to the best of our knowledge, a validated acoustic marker with established discriminative and evaluative properties has not yet been developed for oral tongue cancers. Here we cross-sectionally collected a screening dataset that included acoustic parameters extracted from 3 sustained vowels /ɑ/, /i/, /u/ and binary perceptual outcomes from 12 consonant-vowel syllables. We used a support vector machine with linear kernel function within this dataset to identify the formant centralization ratio (FCR) as a dominant predictor of different perceptual outcomes across gender and syllable. The Acoustic analysis, Perceptual evaluation and Quality of Life assessment (APeQoL) was used to validate the FCR in 33 patients with primary resectable oral tongue cancers. Measurements were taken before (pre-op) and four to six weeks after (post-op) surgery. The speech handicap index (SHI), a speech-specific questionnaire, was also administrated at these time points. Pre-op correlation analysis within the APeQoL revealed overall consistency and a strong correlation between FCR and SHI scores. FCRs also increased significantly with increasing T classification pre-operatively, especially for women. Longitudinally, the main effects of T classification, the extent of resection, and their interaction effects with time (pre-op vs. post-op) on FCRs were all significant. For pre-operative FCR, after merging the two datasets, a cut-off value of 0.970 produced an AUC of 0.861 (95% confidence interval: 0.785-0.938) for T3-4 patients. In sum, this study determined that FCR is an acoustic marker with the potential to detect disease and related speech function in oral tongue cancers. These are preliminary findings that need to be replicated in longitudinal studies and/or larger cohorts.

Patient-Reported Quality of Life After Resection With Primary Closure for Oral Tongue Carcinoma.

OBJECTIVES/HYPOTHESIS: For early-stage oral tongue carcinoma and carcinoma in situ (ESOTCCIS), we evaluated patient-reported quality-of-life (QOL) outcomes following resection with primary closure (R-PC). STUDY DESIGN: Retrospective review at an academic cancer center. METHODS: Thirty-nine ESOTCCIS patients (Tis, T1, T2) who underwent R-PC without radiation completed the University of Washington Quality of Life Questionnaire Version 4 (UW-QOL) at least 6 months since R-PC (mean = 2.39 years; range = 0.5-6.7 years). We compared UW-QOL scores for pain, swallowing, chewing, speech, and taste to established normative population scores. Multivariable regression analysis evaluated factors associated with QOL impairment. RESULTS: ESOTCCIS patients who underwent R-PC in comparison to the normative population reported significantly worse mean speech (87.7 vs. 98, P < .001) and taste (85.6 vs. 95, P = .002) scores and no significant differences in mean pain (91.7 vs. 86, P = .96), swallowing (100 vs. 98, P = .98), chewing (97.4 vs. 94, P = .98) scores. For speech and taste, 59% (23/39) reported no postoperative change from baseline, whereas 41% (16/39) and 35.9% (14/39) reported mild impairment, respectively. Overall, postoperative QOL was reported as good, very good, or outstanding by 87.2% (34/39). Higher American Society of Anesthesiologists class, cT1 compared to CIS, and ventral tongue involvement were independently associated with worse speech. Age < 60 years was independently associated with worse taste. CONCLUSIONS: ESOTCCIS patients who undergo R-PC without radiation can expect long-term swallowing, chewing, and pain to be in the normative range. Although a majority of patients can expect to achieve normative speech and taste outcomes, R-PC carries the risks of mild speech and/or taste impairments. LEVEL OF EVIDENCE: 4 Laryngoscope, 131:312-318, 2021.

Fu's subcutaneous needling for subcutaneous adhesions and scar hyperplasia in the neck region: A case report.

RATIONALE: Lymphadenectomy for tongue cancer in the neck region is often accompanied by local impaired mobility, gland damage, difficult in swallowing, and postoperative complication and seriously affects patients life quality. We reported a case of subcutaneous adhesions and scar hyperplasia in the neck region after lymphadenectomy for tongue lesions accompanied by impaired neck mobility and difficult in swallowing was treated using Fu's subcutaneous needling (FSN) treatment. PATIENT CONCERNS: A 55-year-old male with tongue cancer received surgical intervention with lymphadenectomy 8 years ago was revealed a 15 cm-long curved surgical incision in the neck region and surrounded by numerous scar tissues. DIAGNOSIS: Post-operation subcutaneous adhesions and scar hyperplasia in the neck region after lymphadenectomy was diagnosed. INTERVENTIONS: FSN treatment was performed 2 to 3 times per week for 1 month to sway the affected tightened muscle and dissociate the superficial fascia beneath the scar resulted in a considerable improvement in neck movement. OUTCOMES: The Vancouver Scar Scale (VSS) was as follows: color (M) - 1; vascular distribution (V) - 0, thickness (H) - 2, and flexibility (P) - 4, with a total of 7 points before FSN treatment. The VSS after 1 month of FSN treatment was as follows: M1, V0, H2, and P2, with a total of 5 points. Neck mobility in different directions, i.e., stretching to the back of the neck and laterally bending the neck to the left and/or right side, was improved (P < .05). LESSONS: At present, treatment of chronic scar hyperplasia has certain side effects and limitations. FSN is safe and convenient, with minimal destruction of the superficial fascia, having evident effects of dissociating tissue adhesion under scars and compensating for deficiencies in scar hyperplasia treatment. It can provide new ideas for future treatments.

Impairments in Food Oral Processing in Patients Treated for Tongue Cancer.

Patients surgically treated for oral cancer are affected by several underestimated deglutition disorders risk factors. This study aims to characterize the level of these food oral processing (FOP) impairments in a group of patients treated by surgery for tongue cancer. Twenty-seven consecutive patients surgically treated for tongue cancer were evaluated concerning their chewing capacity (Mastication-test), and responded to questions concerning their capacity to bite, chew and manipulate food with their tongue, and their quality of life. According to the Mastication-test, 16 patients suffered total FOP incapacities (TI group), characterized by high tumor stage, invasive carcinological surgery and necessity of reconstructive surgery; 12 patients were partially or not impaired (PI/NI-group). Tongue movement score and number of dental units were lower in the TI group than in the PI/NI-group. Subjective FOP criteria were clearly impaired in the TI group and correlated with a poor oral health-related quality of life. One year after surgery, there is a decrease in BMI for TI group patients while the PI group patients had a significant increase in BMI. All patients surgically treated for oral cancer suffered FOP impairments, but not with the same severity. Totally impaired subjects are at higher risk from long-term malnutrition. Functional evaluation of FOP should form part of the post-operative follow-up for all patients suffering from tongue cancer, using a quick combined evaluation of chewing efficiency, oral health quality of life and nutritional status.

Circ_0001742 promotes tongue squamous cell carcinoma progression via miR-431-5p/ATF3 axis.

OBJECTIVE: Circular RNAs (circRNAs) have been demonstrated to involve in the development of various cancers. This study aimed to investigate the functions of circ_0001742 on regulating tongue squamous cell carcinoma (TSCC) development and the underlying mechanisms. PATIENTS AND METHODS: The expression of circ_0001742, miR-431-5p and activating transcription factor 3 (ATF3) mRNA was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of epithelial-mesenchymal transition (EMT)-related proteins and ATF3 were measured by Western blot analysis. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry assay were used to evaluate cell proliferation and apoptosis. Besides, Cell migration and invasion were assessed by transwell assay. The relationships between circ_0001742 and miR-431-5p, miR-431-5p and ATF3 were predicted by online software and confirmed by dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and pull-down assay. RESULTS: The expression of circ_0001742 was upregulated in TSCC tissues and cells. Knockdown of circ_0001742 inhibited proliferation, migration, invasion and EMT and induced apoptosis in TSCC cells. Then, miR-431-5p was identified as a target of circ_0001742, and knockdown of miR-431-5p reversed the effects of circ_0001742 knockdown on proliferation, apoptosis, migration, invasion and EMT of TSCC cells. Moreover, miR-431-5p could bind to ATF3, and overexpression of ATF3 rescued the effects mediated by miR-431-5p in TSCC cells. In addition, circ_0001742 regulated ATF3 expression through miR-431-5p. CONCLUSIONS: Our results demonstrated that circ_0001742 plays a tumor-promoting effect in TSCC cells by serving as a competing endogenous RNA (ceRNA) to regulate miR-431-5p/ATF3 axis, which might provide a potential therapeutic target for TSCC.

Atlas-Based Tongue Muscle Correlation Analysis From Tagged and High-Resolution Magnetic Resonance Imaging.

Purpose Intrinsic and extrinsic tongue muscles in healthy and diseased populations vary both in their intra- and intersubject behaviors during speech. Identifying coordination patterns among various tongue muscles can provide insights into speech motor control and help in developing new therapeutic and rehabilitative strategies. Method We present a method to analyze multisubject tongue muscle correlation using motion patterns in speech sound production. Motion of muscles is captured using tagged magnetic resonance imaging and computed using a phase-based deformation extraction algorithm. After being assembled in a common atlas space, motions from multiple subjects are extracted at each individual muscle location based on a manually labeled mask using high-resolution magnetic resonance imaging and a vocal tract atlas. Motion correlation between each muscle pair is computed within each labeled region. The analysis is performed on a population of 16 control subjects and 3 post-partial glossectomy patients. Results The floor-of-mouth (FOM) muscles show reduced correlation comparing to the internal tongue muscles. Patients present a higher amount of overall correlation between all muscles and exercise en bloc movements. Conclusions Correlation matrices in the atlas space show the coordination of tongue muscles in speech sound production. The FOM muscles are weakly correlated with the internal tongue muscles. Patients tend to use FOM muscles more than controls to compensate for their postsurgery function loss.

Submental island flap versus radial forearm free flap for oral tongue reconstruction: a comparison of complications and functional outcomes.

OBJECTIVE: This study compared the complications and functional outcomes of patients with oral tongue cancer who had undergone reconstruction using a submental island flap or a radial forearm free flap. METHOD: Of the 54 patients, 29 underwent reconstruction with a submental island flap and 25 patients with a radial forearm free flap. The complications and outcomes of speech and swallowing were evaluated. RESULTS: In the submental island flap group, all the flaps were successfully transferred with no donor site complications. In the radial forearm free flap group, partial skin graft loss and arm function restriction were recorded. Mean operative time and duration of hospital stay were significantly shorter in the submental island flap group. Speech and swallowing function were comparable between the two groups. There was no significant difference in locoregional recurrence between the groups. CONCLUSION: The submental island flap is reliable and is suitable for oral tongue reconstruction. It has a lower complication incidence when compared to the radial forearm free flap, while maintaining speech and swallowing function.

Gastro-omental free flap for reconstruction of tongue defects.

OBJECTIVES: The purpose of this paper is to report the technique and outcomes of the use of gastro-omental free flap reconstruction in glossectomy defects. MATERIALS AND METHODS: This is a prospective case series of 9 patients of tongue squamous cell carcinoma, who underwent either subtotal or partial glossectomy and reconstruction with gastro-omental free flap. The flap anatomy, surgical technique and the outcomes including the swallowing and speech are presented. RESULTS: Five patients underwent partial glossectomy and 4 had sub-total glossectomy. The median age was 43 years; and the median follow up was 11.4 months. Laparoscopic harvest was done in 8 patients. There was one flap loss. Seven patients underwent postoperative radiotherapy. Functional evaluation was done in 5 patients who were disease free. Four could tolerate soft diet orally, one patient was on liquid to pureed diet. Speech was intelligible in 4. None of the patients had any complications related to laparotomy or laparoscopy. CONCLUSION: Gastro-omental flap provided a secretory mucosal surface and was beneficial in the saliva depleted patients post radiotherapy. The laparoscopic harvest of this flap has minimized donor site morbidity. One patient had a flap loss. Two patients reported superficial ulcerations on the surface, one of them had to undergo surgical debulking to correct it while the other healed with conservative measures. Speech and swallowing outcomes of the reconstructed tongue was good, especially in patients with partial glossectomy. The reconstructed gastric mucosal flaps tolerated the adjuvant radiation well.

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro.

In order to construct an effective and realistic model for tongue squamous cell carcinoma (TSCC) in vitro, the methods were created to produce decellularized tongue extracellular matrix (TEM) which provides functional scaffolds for TSCC construction. TEM provides an in vitro niche for cell growth, differentiation, and cell migration. The microstructures of native extracellular matrix (ECM) and biochemical compositions retained in the decellularized matrix provide tissue-specific niches for anchoring cells. The fabrication of TEM can be realized by deoxyribonuclease (DNase) digestion accompanied with a serious of organic or inorganic pretreatment. This protocol is easy to operate and ensures high efficiency for the decellularization. The TEM showed favorable cytocompatibility for TSCC cells under static or stirred culture conditions, which enables the construction of the TSCC model. A self-made bioreactor was also used for the persistent stirred condition for cell culture. Reconstructed TSCC using TEM showed the characteristics and properties resembling clinical TSCC histopathology, suggesting the potential in TSCC research.

Role of dental hardware in oral cavity squamous cell carcinoma in the low-risk nonsmoker nondrinker population.

BACKGROUND: Oral cavity squamous cell carcinoma (SCC) arising in nonsmokers and nondrinkers remains poorly characterized. We hypothesized that these patients had prior exposure to metallic dental hardware. METHODS: We utilized a questionnaire querying the lifetime oral health status of 54 patients. Demographics and extensive oral health history were collected. RESULTS: The majority of patients (74%) had prior exposure to metallic dental hardware. The younger population with almost exclusively oral tongue cancer had a high prevalence of metallic orthodontic braces (40%) within 15 years before diagnosis. In the 51+ year age group, 82% had crowns, dental implants, and/or dentures with metallic elements. CONCLUSION: Exposure to metallic dental hardware has increased in the past few decades given the rise of orthodontic braces and older adults retaining more teeth. Although this study does not prove a causal relationship between oral cavity SCC and dental hardware, this is a step toward identifying and investigating their role.

Functional evaluation of swallowing in patients with tongue cancer before and after surgery using high-speed continuous magnetic resonance imaging based on T2-weighted sequences.

OBJECTIVES: The aim of this study was to determine the usefulness of evaluating the function of swallowing before and after surgery in patients with tongue cancer by using T2-weighted sequences of high-speed continuous magnetic resonance imaging (HSCMRI). STUDY DESIGN: The imaging findings and related parameters on HSCMRI along with those on routine MRI examinations before and after surgery were examined in 19 patients with tongue cancer. In addition, changes in various parameters during 1 year after surgery were evaluated in 10 patients. RESULTS: In most patients examined, the direction of flow to the esophagus could be seen on HSCMRI before and after surgery. Significant correlations were observed among 4 parameters and in the responses to a dysphagia questionnaire. CONCLUSIONS: The results of the present study suggest that the dynamics of swallowing can be directly visualized on HSCMRI by using 4 parameters that permit the evaluation of changes before and after surgery, and this enables objective evaluation of patients' swallowing complaints.

The Effects of Palate Features and Glossectomy Surgery on /s/ Production.

Purpose: The aims of this article were to determine the effects of hard palate morphology and glossectomy surgery on tongue position and shape during /s/ for patients with small tumors. The first expectation was that laminal /s/ would be more prevalent in patients, than apical, due to reduced tongue tip control after surgery. The second was that patients would hold the tongue more anteriorly than controls to compensate for reduced tongue mass. Method: Three-dimensional tongue volumes were calculated from magnetic resonance imaging for the whole tongue and the portion anterior to the first molar during the /s/ in /əsuk/ for 21 controls and 14 patients. These volumes were used to calculate tongue anteriority and cross-sectional shape. Dental casts were used to measure palate perimeter, height, and width of the hard palate. Results: Palate height correlated with tongue height in controls (p < .05), but not patients. In patients, tongue anteriority correlated negatively with canine width and cross-sectional tongue shape (p < .05). Controls with a high palate favored laminal /s/. Patients preferred laminal /s/ regardless of palate height (p < .01). Conclusions: For controls, hard palate height affected tongue height; a higher palate yielded a higher tongue. For patients, hard palate width affected tongue width; a narrower palate yielded a more anterior tongue. Tongue shape was unaffected by any palate features. Preference for /s/ showed an interaction effect between subject and palate height. Controls with high palates preferred a laminal /s/. All patients preferred a laminal /s/; glossectomy surgery may reduce tongue tip control.

Identification of Key Genes and Pathways in Tongue Squamous Cell Carcinoma Using Bioinformatics Analysis.

BACKGROUND Tongue squamous cell carcinoma (TSCC) is a major type of oral cancers and has remained an intractable cancer over the past decades. The aim of this study was to identify differentially expressed genes (DEGs) during TSCC and reveal their potential mechanisms. MATERIAL AND METHODS The gene expression profiles of GSE13601 were downloaded from the GEO database. The GSE13601 dataset contains 57 samples, including 31 tongue SCC samples and 26 matched normal mucosa samples. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) enrichment analyses were performed; Cytoscape software was used for the protein-protein interaction (PPI) network and module analysis of the DEGs. RESULTS We identified a total of 1,050 upregulated DEGs (uDEGs) and 702 downregulated DEGs (dDEGs) of TSCC. The GO analysis results showed that uDEGs were significantly enriched in the following biological processes (BP): signal transduction, positive or negative regulation of cell proliferation, and negative regulation of cell proliferation. The dDEGs were significantly enriched in the following biological processes: signal transduction, cell adhesion, and apoptotic process. The KEGG pathway analysis showed that uDEGs were enriched in metabolic pathways, pathways in cancer, and PI3K-Akt signaling pathway, while the dDEGs were enriched in focal adhesion and ECM-receptor interaction. The top centrality hub genes RAC1, APP, EGFR, KNG1, AGT, and HRAS were identified from the PPI network. Module analysis revealed that TSCC was associated with significant pathways, including neuroactive ligand-receptor interaction, calcium signaling pathway, and chemokine signaling pathway. CONCLUSIONS The present study identified key genes and signal pathways, which deepen our understanding of the molecular mechanisms of carcinogenesis and development of the disease, and might be used as diagnostic and therapeutic molecular biomarkers for TSCC.

Dose de-escalation to the normal larynx using conformal radiotherapy reduces toxicity while maintaining oncologic outcome for T1/T2 glottic cancer.

We evaluated the efficacy of dose de-escalation to the normal larynx using conformal radiotherapy (CRT) for T1/T2 glottic cancer. For conventional RT (2DRT, n = 38), the laryngeal box received a median equivalent dose in 2 Gy fractions (EQD2) of 66 Gy. For CRT (n = 42; 3D-CRT, 20; intensity-modulated RT, 22), clinical target volume (CTV)1 (gross tumor with a 3-mm margin) and CTV2 (laryngeal box) received median EQD2s of 66.6 Gy and 52.2 Gy, respectively. With a 71-month median follow-up, five-year local control and overall survival rates for 2DRT vs. CRT were 88.1% vs. 95.1% (p = 0.405) and 94.7% vs. 90.7% (p = 0.102), respectively. Grade 2 and 3 pharyngitis rates were 52.6% and 5.3% for 2DRT vs. 35.7% and 2.4% for CRT (p = 0.121). Grade 2 and 3 dermatitis rates were 42.1% and 2.6% for 2DRT vs. 35.7% and 0% for CRT (p = 0.013). The maximum phonation time increased from 12.1 ± 7.1 to 14.0 ± 6.6 seconds after 2DRT (p = 0.375) and from 12.0 ± 5.5 to 13.8 ± 10.1 seconds after CRT (p = 0.313). Fundamental frequency decreased from 150.6 ± 40.3 to 121.9 ± 30.2 Hz after 2DRT (p = 0.039) and from 138.5 ± 31.9 to 126.1 ± 23.7 Hz after CRT (p = 0.058). CRT can effectively de-escalate the normal larynx dose while maintaining oncologic outcome and voice quality.