DAPT Enhances the Apoptosis of Human Tongue Carcinoma Cells

Aim To investigate the effect of DAPT （γ-secretase inhibitor） on the growth of human tongue carcinoma cells and to determine the molecular mechanism to enable the potential application of DAPT to the treatment of tongue carcinoma. Methodology Human tongue carcinoma Tca8113 cells were cultured with DAPT. Cell growth was determined using Indigotic Reduction method. The cell cycle and apoptosis were analyzed by flow cytometry. Real-time PCR and Immuno-Fluorescence （IF） were employed to determine the intracellular expression levels. Results DAPT inhibited the growth of human tongue carcinoma Tca8113 cells by inducing G0-G1 cell cycle arrest and apoptosis, The mRNA levels of Hairy/Enhancer of Split-1 （Hes-1）, a target of Notch activation, were reduced by DAPT in a dose-dependent manner. Coincident with this observation, DAPT induced a dose-dependent promotion of constitutive Caspase-3 in Tca8113 cells. Conclusion DAPT may have a therapeutic value for human tongue carcinoma. Moreover, the effects of DAPT in tumor inhibition may arise partly via the modulation of Notch- 1 and Caspase-3.

Evaluation of the Curative Effect of External Beam Radiotherapy and Brachytherapy for Tongue Carcinoma

OBJECTIVE To evaluate the curative effect of external beam radiothera-py (EBRT) and brachytherapy (BT) for tongue carcinoma. METHODS From 1991 to 2003, 35 patients received EBRT and BT in our department. We analyzed their curative and side effects retrospectively. RESULTS Local control was 80%. The 3-year overall (OS) and disease specific survival (DSS) rates were 75% and 79%. One patient developed metastases. Three patients (9%) developed different late complications. CONCLUSIONS Local regional control, survival, and complications in patients with tongue carcinoma treated by EBRT and BT have been satis-factory.

The activation of adenosine monophosphate–activated protein kinase inhibits the migration of tongue squamous cell carcinoma cells by targeting Claudin-1 via epithelial–mesenchymal transition

Background:The role of Claudin-1 in tongue squamous cell carcinoma(TSCC)metastasis needs further clarification,particularly its impact on cell migration.Herein,our study aims to investigate the role of Claudin-1 in TSCC cell migration and its underlying mechanisms.Methods:36 TSCC tissue samples underwent immunohistochemical staining for Claudin-1.Western blotting and immunofluorescence analyses were conducted to evaluate Claudin-1 expression and distribution in TSCC cells.Claudin-1 knockdown cell lines were established using short hairpin RNA transfection.Migration effects were assessed through wound healing assays.Furthermore,the expression of EMTassociated molecules was measured via western blotting.Results:Claudin-1 expression decreased as TSCC malignancy increased.Adenosine monophosphate–activated protein kinase(AMPK)activation led to increased Claudin-1 expression and membrane translocation,inhibiting TSCC cell migration and epithelial–mesenchymal transition(EMT).Conversely,Claudin-1 knockdown reversed these inhibitory effects on migration and EMT caused by AMPK activation.Conclusions:Our results indicated that AMPK activation suppresses TSCC cell migration by targeting Claudin-1 and EMT pathways.

Acute severe hypokalemia caused by treatment of tongue squamous cell carcinoma with docetaxel and cisplatin:A case report

BACKGROUND The tongue squamous cell carcinoma(TSCC)is an oral malignant tumor arising from the squamous epithelium of the tongue mucosa,characterized by a high malignant degree,invasive growth,early lymph node metastasis,and poor prognosis.Paclitaxel,represented by docetaxel,is now the standard first-line treatment for head and neck squamous cell carcinoma.Docetaxel,which belongs to the class of drugs known as paclitaxel,is an antitumor drug that inhibits cell mitosis and proliferation.Its adverse effects include myelosuppression,hair loss,gastrointestinal reactions,fluid retention,and allergic reactions.However,hypokalemia is rare,most cases are mild or moderate,and severe hypokalemia is seldom reported.symptoms of adverse effects early.It is necessary to be considerate regarding individual differences between patients when selecting chemotherapy regimens and adhere to the principle of individualized treatment.Following multiple cycles of chemotherapy,patients should be aware of the accumulation of toxic side effects and receive blood tests reviewed within 24 hours of completion.It is essential to monitor electrolyte levels in patients suffering from severe gastrointestinal reactions to avoid complications that may result in death.

Current Research Status of MicroRNAs in Squamous Cell Carcinoma of the Tongue

Tongue squamous cell carcinoma (TSCC) is the most invasive type of oral malignant tumor, posing a serious threat to human life and health. Its pathogenesis is complex and has a high degree of malignancy. Recurrence and metastasis often lead to poor prognosis. MicroRNAs are a type of single stranded small molecule RNA with only 18 - 25 nucleotides, which can regulate the expression of various genes and participate in the occurrence and development of tumors. Studies have found that microRNA expression profiling can serve as a reliable and stable biological indicator for early diagnosis and prognosis of tumors. This article provides a review of the research status of MicroRNAs in squamous cell carcinoma of the tongue.

Sternocleidomastoid flap for reconstruction of tongue small cell carcinoma: A case report

BACKGROUND The management of tongue carcinoma is excision and radical neck dissection followed with reconstruction.This is a case report of a patient with tongue squamous cell carcinoma(SCC)who underwent the procedure with sternocleidomastoid(SCM)flap reconstruction.CASE SUMMARY A 52-year-old woman without smoking history complained tongue ulcer since 3 years ago.Based on the histopathological examination,the patient was diagnosed with T2N2M0 right tongue SCC and underwent wide excision of tumor;right mandibular;neck dissection and were reconstructed with SCM flap.CONCLUSION SCC of the tongue requires wide excision and dissection of the neck and mandible if infiltration into the surrounding lymph nodes has been found.The SCM flap reconstruction could be used post-surgery.

Molecular signaling in cancer stem cells of tongue squamous cell carcinoma:Therapeutic implications and challenges

Head and neck squamous cell carcinoma is the seventh most common cancer worldwide with high mortality rates.Amongst oral cavity cancers,tongue carcinoma is a very common and aggressive oral cavity carcinoma.Despite the implementation of a multimodality treatment regime including surgical intervention,chemo-radiation as well as targeted therapy,tongue carcinoma shows a poor overall 5-year survival pattern,which is attributed to therapy resistance and recurrence of the disease.The presence of a rare population,i.e.,cancer stem cells(CSCs)within the tumor,are involved in therapy resistance,recurrence,and distant metastasis that results in poor survival patterns.Therapeutic agents targeting CSCs have been in clinical trials,although they are unable to reach into therapy stage which is due to their failure in trials.A more detailed understanding of the CSCs is essential for identifying efficient targets.Molecular signaling pathways,which are differentially regulated in the CSCs,are one of the promising targets to manipulate the CSCs that would provide an improved outcome.In this review,we summarize the current understanding of molecular signaling associated with the maintenance and regulation of CSCs in tongue squamous cell carcinoma in order to emphasize the need of the hour to get a deeper understanding to unravel novel targets.

Effect of Interferon Regulatory Factor 1 on Proliferation,Invasion and Migration of Tongue Squamous Carcinoma Cells and Its Mechanism

[Objectives]This study was conducted to investigate the effect of interferon regulatory factor on the invasion and migration of tongue squamous carcinoma cells. [Methods]The expression level of IRF1 in tongue squamous carcinoma tissues was detected by real-time quantitative PCR and immunohistochemistry. Plasmids for overexpression and knockdown of IRF1 were constructed. The effects of overexpression and knockdown of IRF1 on the proliferation, invasion and migration of Tca8113 cells were examined in Tca8113 cells. [Results] IRF1 expression was abnormally reduced in tongue squamous carcinoma tissues, and both real-time quantitative PCR and immunohistochemistry showed significantly lower expression than that of paraneoplastic controls. The overexpression and knockdown plasmids of IRF1 were successfully constructed. Growth curve assays showed that overexpression of IRF1 inhibited the proliferation of Tca8113 cells, while knockdown of IRF1 promoted the proliferation of Tca8113 cells. Scratch assay showed that overexpression of IRF1 inhibited the migration of Tca8113 cells, while knockdown of IRF1 promoted the migration of Tca8113 cells. Transwell assay showed that overexpression of IRF1 inhibited the invasion of Tca8113 cells, while knockdown of IRF1 promoted the invasion of Tca8113 cells. [Conclusions] In the development of tongue squamous carcinoma, IRF1 functions as an anti-oncogene, and the expression level of IRF1 was reduced in tongue squamous carcinoma tissues.

Effects of hsa-circ-0001862 on Phenotype of Tongue Squamous Cell Carcinoma Cells

[Objectives]To investigate the molecular mechanism of hsa_circ_0001862 on the proliferation,migration,invasion and apoptosis of tongue squamous cell carcinoma Tca-8113 cells.[Methods]hsa_circ_0001862 plasmid was constructed,and the interaction relationship between hsa_circ_0001862 and miR-23a-3p was verified by dual luciferase reporter gene and qRT-PCR.CCK8 assay,colony formation assay,scratch assay and Transwell assay were used to detect the proliferation,migration and invasion ability of Tca-8113 cells.Western blot was used to detect the expression level of apoptosis-related protein molecules.The effects of hsa_circ_0001862 on the apoptosis of Tca-8113 cells was detected.[Results]hsa_circ_0001862 and miR-23a-3p could interact,and their expression was negatively correlated in tongue squamous cell carcinoma cells.In Tca-8113 cells,hsa_circ_0001862 inhibited cell proliferation,migration,and invasion(P<0.01),and promoted cell apoptosis(P<0.01).[Conclusions]The hsa_circ_0001862 interacts with miR-23a-3p,and hsa_circ_0001862 plays an inhibitory role in the development of tongue squamous cell carcinoma.The hsa_circ_0001862 may be a new biomarker and target for the treatment of tongue squamous cell carcinoma.

Effects of Roundabout 5 on adhesion, invasion and potential motility of human tongue carcinoma Tb cells

Background Roundabout 5 （R5） is a monoclonal antibody which can neutralize the binding of Roundabout 1 （Robol） to Slit2. Oral squamous cell carcinoma angiogenesis was significantly inhibited when R5 blocked slit-robo signaling pathway. However, the effect of R5 on the invasion of tongue cancer cells has not been investigated clearly. Methods In this study, we treated human brain metastasis of tongue cancer cell lines （Tb cells） with R5 at different concentrations, and the control Tb cells were treated with 10 mg/ml immunoglobin G 2b （IgG2b）. The effect of R5 on the proliferation, adhension, invasion and motility of Tb cells was evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide （Ml-r） assay, cell attachment assay on fibronectin （FN）, wound assay and chemotaxis assay, respectively. And gelatin-incorporated sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） was used to investigate the activity of matrix metalloproteinase-2 （MMP2） and matrix metalloproteinase-9 （MMP9）. Results R5 had no effect on the proliferation of Tb cells. However, R5 could significantly inhibit the motility, attachment and chemotaxis of Tb cells to FN, and it could also significantly inhibit the activity of MMP2 and MMP9 in Tb cells. Conclusion R5 can inhibit the adhesion, invasion and motility of human tonaue carcinoma Tb cells.

In vitro effect of iASPP on cell growth of oral tongue squamous cell carcinoma

iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and potential role of iASPP in oral tongue squamous cell carcinoma （OTSCC） have not been addressed. In our study, we detected iASPP expression in OTSCC by immunohistochemistry, iASPP expression is up-regulated in OTSCC tissues. Moreover, in clinical pathology specimens, we found that increased iASPP expression correlates with poor differentiation and lymph node metastasis. Using multicellular tumor spheroids （MTS） and flow cytometry, we demonstrated that iASPP down-regulation arrests OTSCC cells at the G0/G1 phase, induces OTSCC cell apoptosis and inhibits OTSCC cell proliferation. These results indicate that iASPP plays a significant role in the progression of OTSCC and may serve as a biomarker or therapeutic target for OTSCC patients.

OCCULT CERVICAL METASTASIS OF SQUAMOUS CELL CARCINOMA OF TONGUE AMONG CN0 PATIENTS AND ITS TREATMENT

Objective： To explore the treatment of clinically negative neck （CN0） patients with squamous cell carcinoma of the tongue. Methods： 165 CN0 patients with squamous cell carcinoma of the tongue from 1985 to 2002 were investigated retrospectively. Parts of the patients staged at T1, T2 and T3 underwent resection of primary lesion followed by neck observation, and other patients staged above T2 or at T1 but without follow-up were treated with elective neck dissection （END）. All patients were followed up for more than 3 y or until their death. Results： Lymphatic metastasis was identified histologically after operation in 33 of 120 patients treated with END, and 9 of 45 patients treated with resection of primary lesion alone. The overall rate of occult lymphatic metastasis was 25.45%, which increased with the elevating of clinical T stage. The overall rate of neck uncontrolled death was 20.00% for observation group and 5.00% for END group, and significant difference was found between them （P〈0.05）. For T~ patients in the two groups, the rate of neck uncontrolled death was 7.71% and 4.00% respectively, and no significance was found between them （P〉0.05）. When stage T2 and T3 were considered as middle stage together, significant difference （P〈0.05） could be obtained between observation （70.00%） and END group （0%）. Conclusion： The occult metastasis rate of squamous cell carcinoma of tongue increases with the elevating of clinical stage, and elective neck dissection could be considered for NO patients staged over T2 to improve neck control and survival rate; and regional resection alone of primary lesion could be considered for T1N0 patients to improve quality of life if closely followed up is conducted.

Development and validation of novel nomograms to predict survival of patients with tongue squamous cell carcinoma

BACKGROUND There is no unified standard to predict postoperative survival in patients with tongue squamous cell carcinoma(TSCC),hence the urgency to develop a model to accurately predict the prognosis of these patients.AIM To develop and validate nomograms for predicting overall survival(OS)and cancer-specific survival(CSS)of patients with TSCC.METHODS A cohort of 3454 patients with TSCC from the Surveillance,Epidemiology,and End Results(SEER)database was used to develop nomograms;another independent cohort of 203 patients with TSCC from the Department of Oral and Maxillofacial Surgery,First Affiliated Hospital of Zhejiang University School of Medicine,was used for external validation.Univariate and multivariate analyses were performed to identify useful variables for the development of nomograms.The calibration curve,area under the receiver operating characteristic curve(AUC)analysis,concordance index(C-index),net reclassification index(NRI),and decision curve analysis(DCA)were used to assess the calibration,discrimination ability,and clinical utility of the nomograms.RESULTS Eight variables were selected and used to develop nomograms for patients with TSCC.The Cindex(0.741 and 0.757 for OS and CSS in the training cohort and 0.800 and 0.830 in the validation cohort,respectively)and AUC indicated that the discrimination abilities of these nomograms were acceptable.The calibration curves of OS and CSS indicated that the predicted and actual values were consistent in both the training and validation cohorts.The NRI values(training cohort:0.493 and 0.482 for 3-and 5-year OS and 0.424 and 0.402 for 3-and 5-year CSS;validation cohort:0.635 and 0.750 for 3-and 5-year OS and 0.354 and 0.608 for 3-and 5-year CSS,respectively)and DCA results indicated that the nomograms were significantly better than the tumor-node-metastasis staging system in predicting the prognosis of patients with TSCC.CONCLUSION Our nomograms can accurately predict patient prognoses and assist clinicians in improving decision-making concerning patients with TSCC in clinical practice.

Therapeutic effect of AdCMVCD/5-FC system and metabolism of 5-FC in the treatment of human tongue squamous cell carcinoma

To investigate the therapeutic effect and metabolism of 5 fluorocytosine (5 FC ) in human tongue squamous carcinoma cells after treatment with adenovirus medi ated cytosine deaminase (AdCMVCD)/5 FC system Methods Human tongue squamous carcinoma cells (Tca8113 cell line) and its xenografts in BALB/c nude mice were treated with AdCMVCD/5 FC system The killing effect in vitro and bystander effect were detected by microculture tetrazolium (MTT) assay Tumor inhibition effect and histopathological changes were observed in vivo High performance liquid chromatography (HPLC) was performed to determine the m etabolism of 5 FC in vitro and in vivo Results AdCMVCD/5 FC system had strong killing effect and bystander effect on Tca8113 c ells Both condition media and cell extracts showed two peaks identified as 5 FC and 5 fluorouracil (5 FU) by HPLC and a time dependent generation of 5 FU and concomitant time dependent decreases of 5 FC Compared to the control gr oups, mice treated with AdCMVCD/5 FC system demonstrated significant tumor regr ession ( P <0 001); the tumor doubling time prolonged and inhibition rate was 92 62% There were substantial tumor necrotic areas and infiltrative lymphocy tes around necrotic areas in the AdCMVCD/5 FC treated group under light microsc ope There was a significantly low concentration of 5 FC and high concentratio n of 5 FU in tumor tissue, but only 5 FC was found in blood Conclusion AdCMVCD/5 FC suicide gene system had significant in vitro and in vivo anti tum or effect on human tongue squamous cell carcinoma due to convert 5 FC into 5 F U

miR-29b upregulates miR-195 by targeting DNMT3B in tongue squamous cell carcinoma

MicroRNAs play important roles in the devel- opment and progression of various cancers, including tongue squamous cell carcinoma （TSCC）. miR-29b and miR-195 have been reported to be tumor suppressors in TSCC. Here, we investigated the expression of miR-29b and miR- 195 and their relationship in TSCC. Our data showed that miR-29b and miR-195 were significantly downregulated in TSCC com- pared with their matched nonmalignant tissues in 60 paired samples. The level of miR-29b was positively correlated with that of miR-195 in TSCC and the matched nonmalignant tissues. Moreover, miR-29b overexpression induced the demethylation of CpG islands upstream of miR-195 via targeting DNMT3B, leading to the upregulation of miR-195 in TSCC cell lines. Following DNMT3B silencing, the expression of miR-195 was increased and the methylation of CpG islands upstream of miR-195 was reduced. Although overexpression of miR-29b alone significantly increased miR- 195 expression, co-transfection of miR-29b with DNMT3B resulted in no change in miR-195 expression. Taken together, our results demonstrated that miR-29b could upregulate miR- 195 by directly targeting DNMT3B in TSCC. The interaction between miR-29b and miR-195 might provide new insights in developing novel therapeutic approaches of TSCC.

miR-532-3p inhibits the progression of tongue squamous cell carcinoma by targeting podoplanin

Background:The association between miR-532-3p and tongue squamous cell carcinoma(TSCC)has been examined in the literature to improve the survival rate of patients with this tumor.However,further studies are needed to confirm the regulatory roles of this microRNA(miRNA)in TSCC.The objective of this study was to investigate the roles played by and the underlying mechanism used by the miR-532-3p/podoplanin(PDPN)axis in TSCC development.Methods:Western blotting and quantitative real-time reverse transcription-polymerase chain reaction(RT-qPCR)were performed to evaluate the PDPN expression level in TSCC tissues and cells.The proliferative,adhesive,and migratory capabilities of TSCC cells(CAL-27 and CTSC-3)were examined using cell counting kit-8(CCK-8),cell adhesion,and wound-healing assays,respectively.The dual-luciferase reporter(DLR)assay was later conducted to confirm the relationship between miR-532-3p and PDPN.Results:The results indicated that PDPN expression was enriched in TSCC tissues and cells,and that the expression of PDPN was associated with some clinicopathological parameters of TSCC,including lymph node metastasis(P=0.001),tumor-node-metastasis(TNM)staging(P=0.010),and grading(P=0.010).Further analysis also showed that PDPN knockdown inhibited the viability,adhesive ability,and migratory capacity of CAL-27 and CTSC-3 cells,effects that could be reversed by the application of a miR-532-3p inhibitor.Additionally,PDPN was found to be a direct target of miR-532-3p.Conclusions:This research suggested that by targeting PDPN,miR-532-3p could inhibit cell proliferation viability,adhesion,and migration in TSCC.Findings also revealed that the miR-532-3p/PDPN axis might provide more insights into the prognosis and treatment of TSCC.

Effects of ATRA, Acitretin and Tazarotene on Growth and Apoptosis of Tca8113 Cells

Summary：To investigate the effects of ATRA, acitretin and tazarotene on the growth and apoptosis of human tongue squamous cell carcinoma cell line Tca8113. The effect of retinoids on growth of Tca8113 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis with double staining with Annexin V-FITC and PI, and active caspase-3 analysis with the staining of FITC-conjugated monoclonal rabbit anli-active caspase-3 antibody were made by flow cytometer. Streptavidin-biotin complex （SABC） immunocytochemical assays were employed for the detections of Bax/Bcl-2 proteins expressions. Our results showed that the retinoids inhibited growth of Tca8113 cells in a dose-and time-dependent manner with maximal inhibition 24 h after treatment of 10 5 mol/L. 10^-5 mol/L retinoids altered cell cycle distribution of Tca8113 cells, revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. 10^-5 mol/L retinoids significantly induced apoptosis of Tca8113 cells （all P〈0.05）, elevated the cells population with detectable active caspase-3 （P〈 0.05 for all）, increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly （all P〈0.05）. Acitretin played a most prominent role among the retinoids. It is concluded that the inhibition of cell cycle progress of Tca8113 cells by ATRA, acitretin and tazarotene is one of the possible mechanisms for proliferation arrest of TcaS113 cells elicited by the retinoids. The retinoids mediate apoptosis in TcaS113 cells that may be caspase-dependent through mitochondria pathway. High concentration retinoids inhibit growth of Tca8113 cells in vitro by interfering with proliferation and inducing apoptosis of cells. Acitretin may be an alternative medicine for the prevention and treatment of tongue squamous cell carcinoma.

Squamous cell carcinoma of tongue 18 years after renal transplantation:a case report

Solid organ transplant recipients are at increased risk of developing malignancies,even decades after transplant,due to the prolonged use of immunosuppressant drugs.A 35-year-old male underwent renal transplant for end stage renal disease 18 years previously and was on immunosuppressive drugs since that time and was on regular follow up.In 2016,he developed a squamous cell carcinoma of tongue,which was operated and adjuvant radiation therapy was given.The patient is currently on follow up and asymptomatic.Though squamous cell carcinoma of tongue is a relatively common malignancy in the general population,it is very rare in transplant recipients.Hence,such patients require longer follow-up,active surveillance,and screening for early diagnosis and prompt treatment of premalignant and malignant conditions.

Flowcytometry Analysis in 67 Squamous Cell Carcinoma of the Tongue.

Flowcytometry (FCM) was used to measure the DNA content in 67 cases of squamous cell carcinoma of the tongue (SCCT). The relationship of FCM results were compared with clinical and pathological parameters. The results showed