Tongue squamous cell carcinoma is highly malignant and has a poor prognosis. In this study, we aimed to combine whole-genome sequencing, whole-genome methylation, and whole-transcriptome analyses to understand the molecular mechanisms of tongue squamous cell carcinoma better. Oral tongue squamous cell carcinoma and adjacent normal tissues from five patients with tongue squamous cell carcinoma were included as five paired samples. After multi-omics sequencing, differentially methylated intervals, methylated loop sites, methylated promoters, and transcripts were screened for variation in all paired samples. Correlations were analyzed to determine biological processes in tongue squamous cell carcinoma. We found five mutated methylation promoters that were significantly associated with mRNA and lncRNA expression levels. Functional annotation of these transcripts revealed their involvement in triggering the mitogen-activated protein kinase cascade, which is associated with cancer progression and the development of drug resistance during treatment. The prognostic signature models constructed based on WDR81 and HNRNPH1 and combined clinical phenotype-gene prognostic signature models showed high predictive efficacy and can be applied to predict patient prognostic risk in clinical settings. We identified biological processes in tongue squamous cell carcinoma that are initiated by mutations in the methylation promoter and are associated with the expression levels of specific mRNAs and lncRNAs. Collectively, changes in transcript levels affect the prognosis of tongue squamous cell carcinoma patients.
Humans ; Biomarkers, Tumor ; Nerve Tissue Proteins ; Prognosis ; Squamous Cell Carcinoma of Head and Neck/pathology* ; Tongue Neoplasms/pathology*

Objective: To investigate the efficacies of different forms of free radial collateral artery perforator flaps in repairing the defects after oral tumor surgeries. Methods: From May 2016 to March 2021, 28 patients (22 males, 6 females, aged 35-62 years) with oral tumors admitted by Hunan Cancer Hospital received the reconstructive surgeries with the free radial collateral artery perforator flaps after removal of oral tumors, including 24 cases of tongue cancer (11 cases of tongue marginal cancer, 9 cases of tongue belly cancer and 4 cases of tongue cancer involved in the floor of the mouth) and 4 cases of buccal and oral cancer. Four forms of radial collateral artery perforator flaps were used: single perforator flaps for 6 cases, double perforators flaps for 7 cases, flaps without perforator visualization for 10 cases and chimeric perforator myocutaneous flaps for 5 cases. The recipient vessels were the superior thyroid artery and superior thyroid vein, and if second concomitant vein available, it was anastomosed with internal jugular vein in end-to-side fashion. SPSS 20.0 statistical software was used to analyze the data. Results: The mean length of flaps was (9.7±0.4) cm, mean width (4.4±0.3) cm and mean thickness (1.1±0.4) cm. The mean length of the vascular pedicles was (7.1±0.6)cm (6.0-8.0 cm), the mean diameter of the radial accessory arteries was (1.1±0.3)mm (0.8-1.3 mm). Eleven cases(39.3%) had respectively one accompanying vein and 17 cases(60.7%) had respectively two accompanying veins, with the mean diameter of (1.1±0.3) mm (0.8-1.3 mm). All the 28 flaps survived, the donor and recipient wounds healed in one stage, the appearances of the flaps were satisfactory, only linear scars remained in the donor sites, and the upper arm functions were not significantly affected. Follow up for 12-43 months showed that the flaps were soft with partially mucosalization, the reconstructed tongue and buccal cavity were in good shape, and the swallowing and language functions were satisfactory. The swallowing and language functions were retained to the greatest extent in 3 cases with near total tongue resection, although the functions were still significantly affected. There was no local recurrence of the tumor during follow-up. One case had regional lymph node metastasis, and further lymph node dissection and comprehensive treatment were performed, with satisfactory outcomes. Conclusions: The vascular pedicle of the radial collateral artery perforator flap has a constant anatomy, which can be prepared in different forms to improve the safety of the operation and minimize the donor site damage. It is an ideal choice for the repair of small and medium-sized defects after oral tumor surgery.
Male ; Female ; Humans ; Perforator Flap/transplantation* ; Plastic Surgery Procedures ; Tongue Neoplasms/surgery* ; Arm/surgery* ; Mouth Neoplasms/surgery* ; Arteries ; Skin Transplantation ; Treatment Outcome

Malignant peripheral nerve sheath tumor (MPNST) is a rare neurogenic malignant tumor. MPNST has aty-pical clinical symptoms and imaging presentations, difficult diagnosis, a high degree of malignancy, and poor prognosis. It usually occurs in the trunk, approximately 20% in the head and neck, and rarely in the mouth. This paper reports a case of MPNST of the tongue. A summary of the clinical features, diagnosis, and treatment of MPNST is presented in combination with a literature review to provide a reference for the diagnosis and treatment of this disease.
Humans ; Nerve Sheath Neoplasms/pathology* ; Neurofibrosarcoma ; Tongue/pathology*

OBJECTIVES: Tongue squamous cell carcinoma (TSCC) is a common cancer in the oral and maxillofacial region, which seriously endangers people's life and health.Heterogeneous nuclear ribonucleoprotein A2/B1(hnRNP A2/B1) is an RNA-binding protein that regulates the expression of a variety of genes and participates in the occurrence and development of a variety of cancers. This study aims to investigate the role of hnRNP A2/B1 in TSCC progression. METHODS: The differential expression of hnRNP A2/B1 in oral squamous cell carcinoma (OSCC) and normal oral mucosa cells and tissues was analyzed based on the gene expression profiles of GSE146483 and GSE85195 in the Gene Expression Omnibus (GEO) database. The correlation between hnRNP A2/B1 expression and disease-free survival of TSCC patients was analyzed based on TSCC related chip of GSE4676. TSCC cancer and paracancerous tissue samples of 30 patients were collected in Hunan Cancer Hospital from July to December 2021. Real-time RT-PCR and Western blotting were used to verify the mRNA and protein expression of hnRNP A2/B1 in TSCC patients'samples, respectively. Human TSCC Tca-8113 cells were transfected with hnRNP A2/B1 empty vector (a sh-NC group), knockdown plasmid (a sh-hnRNP A2/B1 group), empty vector overexpression plasmid (an OE-NC group) and overexpression plasmid (an OE-hnRNP A2/B1 group), respectively. The knockdown or overexpression efficiency of hnRNP A2/B1 was detected by Western blotting. The proliferation activity of Tca-8113 cells was detected by cell counting kit-8 (CCK-8), and the apoptosis rate of Tca-8113 cells was detected by flow cytometry. RESULTS: Based on the analysis of OSCC-related chips of GSE146483 and GSE85195 in the GEO database, it was found that hnRNP A2/B1 was differentially expressed in the OSCC and normal oral mucosa cells and tissues (all P<0.01). Meanwhile, the analysis of TSCC related chip GSE4676 confirmed that the expression of hnRNP A2/B1 was negatively correlated with the disease-free survival of TSCC patients (P=0.006). The results of real-time RT-PCR and Western blotting showed that the relative expression levels of hnRNP A2/B1 mRNA and protein in TSCC tissues were significantly up-regulated compared with those in adjacent tissues (all P<0.01). The results of Western blotting showed that the expression level of hnRNP A2/B1 in Tca-8113 cells was significantly inhibited or promoted after knockdown or overexpression of hnRNP A2/B1 (all P<0.01). The results of CCK-8 and flow cytometry showed that inhibition of hnRNP A2/B1 expression in Tca-8113 cells reduced cell proliferation activity (P<0.05) and increased cell apoptic rate (P<0.01). Overexpression of hnRNP A2/B1 in Tca-8113 cells significantly increased cell proliferation (P<0.05) and decreased cell apoptosis (P<0.01). CONCLUSIONS HnRNP A2/B1 is a key factor regulating the proliferation and apoptosis of TSCC cells. Inhibition of hnRNP A2/B1 expression can reduce the proliferation activity of TSCC cells and promote the apoptosis of TSCC cells.
Humans ; Carcinoma, Squamous Cell/genetics* ; Sincalide/metabolism* ; Tongue Neoplasms/genetics* ; Mouth Neoplasms ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B/metabolism* ; RNA, Messenger ; Tongue/metabolism* ; Cell Line, Tumor

OBJECTIVE: To evaluate the inhibitory effect of cordycepin on oral cancer xenograft in nude mice and explore the underlying mechanisms. METHODS: Sixteen BALB/c mice bearing subcutaneous human tongue squamous cell carcinoma (TSCC) TCA-8113 cell xenografts were randomized into model group and cordycepin treatment group for daily treatment with saline and cordycepin for 4 weeks. After the treatment, the tumor xenografts were dissected and weighed to assess the tumor inhibition rate. Histological changes in the heart, spleen, liver, kidney, and lung of the mice were evaluated with HE staining, and tumor cell apoptosis was examined using TUNEL staining; The expressions of Bax, Bcl-2, GRP78, CHOP, and caspase-12 in the xenografts were detected using RT-qPCR and Western blotting. RESULTS: Cordycepin treatment resulted in a tumor inhibition rate of 56.09% in the nude mouse models, induced obvious changes in tumor cell morphology and significantly enhanced apoptotic death of the tumor cells without causing pathological changes in the vital organs. Cordycepin treatment also significantly reduced Bcl-2 expression (P < 0.05) and increased Bax, GRP78, CHOP, and caspase-12 expressions at both the RNA and protein levels in the tumor tissues. CONCLUSION Cordycepin treatment can induce apoptotic death of TCA-8113 cell xenografts in nude mice via the endogenous mitochondrial pathway and endoplasmic reticulum stress pathways.
Humans ; Animals ; Mice ; Carcinoma, Squamous Cell/drug therapy* ; Heterografts ; Mice, Nude ; Tongue Neoplasms/drug therapy* ; Cordyceps ; Caspase 12 ; Endoplasmic Reticulum Chaperone BiP ; bcl-2-Associated X Protein ; Tongue

In contrast to the well-established genomic 5-methylcytosine (5mC), the existence of N⁶-methyladenine (6 mA) in eukaryotic genomes was discovered only recently. Initial studies found that it was actively regulated in cancer cells, suggesting its involvement in the process of carcinogenesis. However, the contribution of 6 mA in tongue squamous cell carcinoma (TSCC) still remains uncharacterized. In this study, a pan-cancer type analysis was first performed, which revealed enhanced 6 mA metabolism in diverse cancer types. The study was then focused on the regulation of 6 mA metabolism, as well as its effects on TSCC cells. To these aspects, genome 6 mA level was found greatly increased in TSCC tissues and cultured cells. By knocking down 6 mA methylases N6AMT1 and METTL4, the level of genomic 6 mA was decreased in TSCC cells. This led to suppressed colony formation and cell migration. By contrast, knockdown of 6 mA demethylase ALKBH1 resulted in an increased 6 mA level, enhanced colony formation, and cell migration. Further study suggested that regulation of the NF-κB pathway might contribute to the enhanced migration of TSCC cells. Therefore, in the case of TSCC, we have shown that genomic 6 mA modification is involved in the proliferation and migration of cancer cells.
AlkB Homolog 1, Histone H2a Dioxygenase/metabolism* ; Carcinoma, Squamous Cell/pathology* ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Site-Specific DNA-Methyltransferase (Adenine-Specific)/metabolism* ; Tongue Neoplasms/metabolism*

Objective: To investigate the efficacy of anterolateral thigh flap preforming tongue in patients with total glossectomy. Methods: A total of 27 patients with tongue cancer who underwent total glossectomy, neck lymph node dissection and anterolateral thigh flap transfer were collected from January 2019 to April 2021 in the Department of Oral and Maxillofacial Surgery at the Second Xiangya Hospital. All patients were males, the age ranged from 35-73 years. The patients were divided into experimental (14 cases) and control (13 cases) groups, based on whether the tongue was reconstructed. The clinical parameters of two groups were analyzed by independent sample t test or Fisher exact probability method. Results: The success rate of free flap was 100%, of the patients, 2 patients had cervical hematoma and 1 patient had wound infection postoperatively. There was no difference in speech (6.69±3.42 vs. 5.50±3.01, t=0.96, P=0.346) or swallowing (χ²=0.46, P=0.793) function between two groups at 1 month after surgery. However, the speech (24.94±7.43 vs. 18.44±6.30, t=2.48, P=0.020) and swallowing (χ²=6.97, P=0.008) functions in experimental group were significantly better than those in control group. No case was lost to follow-up. All patients were extubated after operation, with average time of 7.2 days in the experimental group and 7.7 days in the control group. The overall survival rate was 71.4% in the experimental group and 61.5% in the control group. Conclusion: The use of anterolateral thigh flap preforming tongue can improve the speech and swallowing functions in patients with total glossectomy and offer a novel method for tongue construction.
Male ; Humans ; Adult ; Middle Aged ; Aged ; Female ; Glossectomy ; Thigh ; Tongue ; Tongue Neoplasms ; Free Tissue Flaps

Objective: To explore the action mechanism of hsa_circ_0000231 in the occurrence and development of tongue squamous cell carcinoma (TSCC). Methods: Tissue samples of 60 TSCC patients were examined. The patients, including 32 males and 28 females, aged from 36 to 84 years old, underwent surgery in the Affiliated Hospital of Nantong University and Affiliated Tumor Hospital of Nantong University from December 2014 to December 2017. Saliva samples were obtained from healthy volunteers (5 males and 5 females, aged from 40 to 75 years old) and 10 TSCC patients. The TSCC cell lines (CAL-27, Tca-8113 and HN-4) were used. The expression levels of hsa_circ_0000231 in 60 pairs of freshly matched TSCC and para-carcinoma tissue samples, 10 pairs of saliva samples and 3 TSCC cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR). hsa_circ_0000231 gene interference and lentiviral transfection were constructed, hsa_circ_0000231 in TSCC cell lines CAL-27 and Tca-8113 was knocked down, and the expressions of hsa_circ_0000231 in hsa_circ_0000231 interference group (sh-circ) and no-load lentivirus group (negative control) were tested with qRT-PCR. Cells with the highest knock-down efficiency were selected for CCK-8 test, colony formation assay, transwell invasion assay and scratch assay. The expressions of EMT-related proteins including E-cadherin, snail protein, N-cadherin and vimentin and proteins related to Wnt/β-catenin signaling pathway including β-catenin, C-myc, Bcl-2, MMP-9 and Cyclin D1 were measured by western blot. After TSCC cells in the interference group were co-cultured with Wnt/β-catenin pathway activator LiCl, the expressions of above proteins were re-measured by western blot. TSCC cells in interference group and control group were subcutaneously injected into nude mice to compare the effect of hsa_circ_0000231 knockdown on the growths of the tumors grafted subcutaneously in the nude mice. Statistical analysis software 25.0 was used for data analysis, and t-test or chi-square test was used for comparison between groups. Results: hsa_circ_0000231 was highly expressed in the tissue and saliva samples of TSCC patients and cell lines CAL-27, Tca-8113 and HN-4, but lowly expressed in paired para-carcinoma tissues, saliva samples of healthy people and normal human oral keratinocytes (all P<0.05). Log-rank univariate analysis showed that hsa_circ_0000231 expression level, tumor differentiation degree and T stage were related to the survival of TSCC patients (all P<0.05). Multivariate Cox risk regression model analysis suggested that hsa_circ_0000231 expression level (χ²=5.77,P=0.016) and T stage (χ²=5.27,P=0.029) were independent factors for the poor prognosis of TSCC patients. Western blot showed the expressions of snail protein, N-cadherin and vimentin were down-regulated, but E-cadherin was up-regulated in interference group compared with control group. In interference group, the expressions of β-catenin, C-myc, Bcl-2, MMP-9 and CyclinD1 were down-regulated, which were reversed after TSCC cells were co-cultured with LiCl. The knockdown of hsa_circ_0000231 reduced the proliferation, invasion and metastasis abilities of CAL-27 and Tca-8113 cells, which were reversed after TSCC cells were co-cultured with LiCl. The growth rate and volume of the tumors grafted subcutaneously in interference group using LiCl were greater than those in negative control group. Conclusion: hsa_circ_0000231 is an independent prognostic factor of TSCC. Highly expressed hsa_circ_0000231 can promote the proliferation, invasion and metastasis of TSCC cells.
Male ; Animals ; Mice ; Female ; Humans ; Adult ; Middle Aged ; Aged ; Aged, 80 and over ; Tongue Neoplasms ; Wnt Signaling Pathway/genetics* ; Carcinoma, Squamous Cell/genetics* ; beta Catenin/metabolism* ; Mice, Nude ; Vimentin ; Matrix Metalloproteinase 9/metabolism* ; RNA, Circular ; Gene Expression Regulation, Neoplastic ; Cell Proliferation/genetics* ; Cadherins/genetics* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Tongue

OBJECTIVE: To examine the correlation of CCBE1 expression in adjacent tissues of tongue squamous cell carcinoma (TSCC) with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and survival outcomes of the patients. METHODS: Lymphatic vessel density was quantified in pericancerous tissue sections of 44 cases of cT_1-2N₀ TSCC using D2-40 as the lymphatic vessel endothelial marker for calibration and counting of the lymphatic vessels. Of these 44 cases, 22 showed a relatively low lymphatic vessel density (group A) and the other 22 had a high lymphatic vessel density (group B), and the expression levels of CCBE1 in the adjacent tissues determined using immunohistochemistry, immunofluorescence assay and Western blotting were compared between the two groups. The expression level of CCBE1 was also measured in another 90 patients with TSCC using immunohistochemistry, and all the patients were followed up for their survival outcomes. RESULTS: Immunohistochemistry and Western blotting showed a significantly lower rate of high CCBE1 expression in group A than in group B (P < 0.05). Immunofluorescence assay showed co-localization of CCBE1 and D2-40 in the adjacent tissues of TSCC. In the 90 TSCC patients with complete follow-up data, a high expression of CCBE1 was found to correlate with lymph node metastasis and a poor 5-year survival outcomes of the patients (P < 0.05). CONCLUSION A high expression of CCBE1 in the adjacent tissues of TSCC is closely related with pericancerous lymphatic vessel proliferation, cervical lymph node metastasis and a poor 5-year survival of the patients, suggesting the value of CCBE1 as a potential prognostic predictor for TSCC.
Humans ; Tongue Neoplasms/pathology* ; Carcinoma, Squamous Cell/metabolism* ; Lymphatic Metastasis ; Prognosis ; Lymphatic Vessels/pathology* ; Cell Proliferation ; Tongue/pathology* ; Calcium-Binding Proteins/metabolism* ; Tumor Suppressor Proteins/metabolism*

The annual incidence of oral cancer in the world is more than 300 000, and the five-year survival rate is 50%~60%. Every year, 145 400 people died due to tongue cancer, of which tongue cancer accounts for nearly 40%. Although tongue cancer has tumor heterogeneity and individual differences in prognosis, surgery is still the first choice for the treatment of tongue cancer. The effect of tongue cancer surgery can directly determine the survival time of patients. The defect caused by tongue cancer surgery seriously affects the patients' physical functions such as appearance, language, chewing and swallowing. The surgical treatment of tongue cancer with functional reconstruction can improve the quality of life of patients. In the past few decades, genomics has enhanced understanding of tongue cancer. The preclinical tumor model preserves the tumor heterogeneity, which has a great application prospect in the discovery of tumor biomarkers and the clinical translation of drugs. Many advances have been made in the diagnosis and treatment of tongue cancer, but there are still many controversies in clinical practice. Therefore, this expert consensus summarizes the progress and controversial hot spots of surgical diagnosis and treatment of tongue cancer, mainly including preoperative diagnosis and evaluation, surgical treatment points and postoperative functional rehabilitation.
Carcinoma, Squamous Cell/surgery* ; Consensus ; Humans ; Quality of Life ; Plastic Surgery Procedures/methods* ; Tongue/surgery* ; Tongue Neoplasms/surgery*