TMEM16A inhibits autophagy and promotes the invasion of hypopharyngeal squamous cell carcinoma through mTOR pathway.

Previous studies have indicated that transmembrane protein 16A (TMEM16A) plays a crucial role in the pathogenesis and progression of various tumors by influencing multiple signaling pathways. However, the role of TMEM16A in regulating autophagy via the mammalian target of rapamycin (mTOR) pathway and its impact on the development of hypopharyngeal squamous cell carcinoma (HSCC) remain unclear. Immunohistochemistry and western blotting were used to assess the expression of TMEM16A in HSCC tissues and metastatic lymph nodes. Manipulation of TMEM16A expression levels was achieved in the FaDu cell line through overexpression or knockdown, followed by assessment of its biological effects using cell colony formation, wound healing, transwell, and invasion assays. Additionally, apoptosis and autophagy-related proteins, as well as autophagosome formation, were evaluated through western blotting, transmission electron microscopy, and immunofluorescence following TMEM16A knockdown or overexpression in FaDu cells. Our study revealed significantly elevated levels of TMEM16A in both HSCC tissues and metastatic lymph nodes compared to normal tissues. In vitro experiments demonstrated that silencing TMEM16A led to a notable suppression of HSCC cell proliferation, invasion, and migration. Furthermore, TMEM16A silencing effectively inhibited tumor growth in xenografted mice. Subsequent investigations indicated that knockdown of TMEM16A in HSCC cells could suppress mTOR activation, thereby triggering autophagic cell death by upregulating sequestosome-1 (SQSTM1/P62) and microtubule-associated protein light chain 3 II (LC3II). This study highlights the crucial role of TMEM16A in modulating autophagy in HSCC, suggesting its potential as a therapeutic target for the treatment of this malignancy.

m6A methylation reader IGF2BP2 activates endothelial cells to promote angiogenesis and metastasis of lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) is a common type of lung cancer with a high risk of metastasis, but the exact molecular mechanisms of metastasis are not yet understood. METHODS: This study acquired single-cell transcriptomics profiling of 11 distal normal lung tissues, 11 primary LUAD tissues, and 4 metastatic LUAD tissues from the GSE131907 dataset. The lung multicellular ecosystems were characterized at a single-cell resolution, and the potential mechanisms underlying angiogenesis and metastasis of LUAD were explored. RESULTS: We constructed a global single-cell landscape of 93,610 cells from primary and metastatic LUAD and found that IGF2BP2 was specifically expressed both in a LUAD cell subpopulation (termed as LUAD_IGF2BP2), and an endothelial cell subpopulation (termed as En_IGF2BP2). The LUAD_IGF2BP2 subpopulation progressively formed and dominated the ecology of metastatic LUAD during metastatic evolution. IGF2BP2 was preferentially secreted by exosomes in the LUAD_IGF2BP2 subpopulation, which was absorbed by the En_IGF2BP2 subpopulation in the tumor microenvironment. Subsequently, IGF2BP2 improved the RNA stability of FLT4 through m6A modification, thereby activating the PI3K-Akt signaling pathway, and eventually promoting angiogenesis and metastasis. Analysis of clinical data showed that IGF2BP2 was linked with poor overall survival and relapse-free survival for LUAD patients. CONCLUSIONS: Overall, these findings provide a novel insight into the multicellular ecosystems of primary and metastatic LUAD, and demonstrate that a specific LUAD_IGF2BP2 subpopulation is a key orchestrator promoting angiogenesis and metastasis, with implications for the gene regulatory mechanisms of LUAD metastatic evolution, representing themselves as potential antiangiogenic targets.

Evaluating hypopharyngeal carcinoma using narrow band imaging and oxygen-injected laryngoscope: New technique.

OBJECTIVE: To evaluate the diagnostic value of narrow band imaging (NBI) endoscopic classification for hypopharyngeal lesions and to lay the groundwork for practical applications of oxygen-injected laryngoscope for hypopharyngeal carcinoma (HC). METHODS: A total of 140 subjects with suspected 146 hypopharyngeal lesions were selected for pathological examination. Subsequently, NBI and white light imaging (WLI) endoscopy were performed to observe and classify lesions into 7 types according to our modified NBI classification. Pathological results were used as the gold standard to assess the diagnostic value of the NBI classification. The value of oxygen-injected laryngoscope for accurate assessment of lesion extension was evaluated based on the exposure of hypopharyngeal lesions before and after use. RESULTS: The accuracy, sensitivity, and negative predictive value of NBI endoscopy in diagnosing hypopharyngeal lesions were 95.9 %, 96.7 %, and 84.6 %, respectively, which were higher than those of WLI mode (p < 0.05). NBI endoscopy was more accurate than WLI in diagnosing malignant lesions (p < 0.05), especially for high-grade dysplasia (HGD) (p < 0.05). There was remarkable consistency between NBI classification and pathological results (Kappa = 0.855). Type Va and type Vb-c accounted for 72.7 % and 92.8 % of HGD and invasive carcinoma, respectively. Moreover, the oxygen-injected laryngoscope was found to provide a more accurate assessment of HC extension (P < 0.001). CONCLUSION: We propose a more appropriate NBI endoscopic classification for hypopharyngeal lesions, which can effectively improve diagnostic accuracy, especially for the early diagnosis of hypopharyngeal cancer. Moreover, the application of oxygen-injected laryngoscope is essential for the accurate assessment of HC and has a high clinical utility.

Applying a nomogram based on preoperative CT to predict early recurrence of laryngeal squamous cell carcinoma after surgery.

PURPOSE: To identify the value of a computed tomography (CT)-based radiomics model to predict probability of early recurrence (ER) in patients diagnosed with laryngeal squamous cell carcinoma (LSCC) after surgery. MATERIALS AND METHOD: Pre-operative CT scans of 140 LSCC patients treated by surgery are reviewed and selected. These patients are randomly split into the training set (n = 97) and test set (n = 43). The regions of interest of each patient were delineated manually by two senior radiologists. Radiomics features are extracted from CT images acquired in non-enhanced, arterial, and venous phases. Variance threshold, one-way ANOVA, and least absolute shrinkage and selection operator algorithm are used for feature selection. Then, radiomics models are built with five algorithms namely, k-nearest neighbor (KNN), logistic regression (LR), linear support vector machine (LSVM), radial basis function SVM (RSVM), and polynomial SVM (PSVM). Clinical factors are selected using univariate and multivariate logistic regressions. Last, a radiomics nomogram incorporating the radiomics signature and clinical factors is built to predict ER and its efficiency is evaluated by receiver operating characteristic (ROC) curve and calibration curve. Decision curve analysis (DCA) is also used to evaluate clinical usefulness. RESULTS: Four features are remarkably associated with ER in patients with LSCC. Applying to test set, the area under the ROC curves (AUCs) of KNN, LR, LSVM, RSVM, and PSVM are 0.936, 0.855, 0.845, 0.829, and 0.794, respectively. The radiomics nomogram shows better discrimination (with AUC: 0.939, 95% CI: 0.867-0.989) than the best radiomics model and the clinical model. Predicted and actual ERs in the calibration curves are in good agreement. DCA shows that the radiomics nomogram is clinically useful. CONCLUSION: The radiomics nomogram, as a noninvasive prediction tool, exhibits favorable performance for ER prediction of LSCC patients after surgery.

miR-146a enhances regulatory T-cell differentiation and function in allergic rhinitis by targeting STAT5b.

BACKGROUND: MicroRNA (miR)-146a, as an important immune regulatory factor with an anti-inflammatory effect, plays a crucial role in regulatory T-cell (Tregs) differentiation and function in allergic rhinitis (AR). The present study aimed to investigate the regulatory mechanism employed by miR-146a to control Treg differentiation and function in AR. METHODS: Expression of miR-146a and STAT5b in peripheral blood mononuclear cells (PBMCs) and nasal mucosa from patients with AR was detected by qPCR and Western blotting. Tregs were quantified by flow cytometry in miR-146a knockdown or STAT5b knockdown PBMCs. FOXP3, IL-10, and TGF-ß levels were detected by Western blotting or ELISA in miR-146a knockdown or STAT5b overexpressing PBMCs, as well as in STAT5b knockdown PBMCs overexpressing miR-146a. The effect of miR-146a on STAT5b was observed by luciferase assay and knockdown experiments. RESULTS: Levels of miR146a and STAT5b in the nasal mucosa or PBMCs were significantly lower in the AR group than in the control group. There were significantly fewer Tregs in miR-146a knockdown or STAT5b knockdown PBMCs compared to control PBMCs. Expression of FOXP3, IL-10, and TGF-ß was decreased in the miR-146a knockdown group but increased in the STAT5b overexpression group. In contrast, miR-146a overexpression increased the levels of these factors, but knockdown of STAT5b significantly inhibited this effect. Luciferase assay and knockdown experiments showed that miR-146a bound directly to STAT5b. CONCLUSIONS: miR-146a enhances Treg differentiation and function in AR by positively targeting STAT5b.

Radiomics Model for Predicting TP53 Status Using CT and Machine Learning Approach in Laryngeal Squamous Cell Carcinoma.

Objective: We aim to establish and validate computed tomography (CT)-based radiomics model for predicting TP53 status in patients with laryngeal squamous cell carcinoma (LSCC). Methods: We divided all patients into a training set 1 (n=66) and a testing set 1 (n=30) to establish and validate radiomics model to predict TP53. Radiomics features were selected by analysis of variance (ANOVA) and the least absolute shrinkage and selection operator (Lasso) regression analysis. Five radiomics models were established by using K-Nearest Neighbor, logistics regressive, linear-support vector machine (SVM), gaussian-SVM, and polynomial-SVM in training set 1. We also divided all patients into a training set 2 and a testing set 2 according to different CT equipment to establish and evaluate the stability of the radiomics models. Results: After ANOVA and subsequent Lasso regression analysis, 22 radiomics features were selected to build the radiomics model in training set 1. The radiomics model based on linear-SVM has the best predictive performance of the five models, and the area under the receiver operating characteristic curve in training set 1 and testing set 1 were 0.831(95% confidence interval [CI] 0.692-0.970) and 0.797(95% CI 0.632-0.957) respectively. The specificity, sensitivity, and accuracy were 0.971(95% CI 0.834-0.999), 0.714(95% CI 0.535-0.848), and 0.843(95% CI 0.657-0.928) in training set 1 and 0.750(95% CI 0.500-0.938), 0.786(95% CI 0.571-1.000), and 0.667(95% CI 0.467-0.720) in testing set 1, respectively. In addition, the radiomics model also achieved stable prediction results even in different CT equipment. Decision curve analysis showed that the radiomics model for predicting TP53 status could benefit LSCC patients. Conclusion: We developed and validated a relatively optimal radiomics model for TP53 status prediction by trying five different machine learning methods in patients with LSCC. It shown great potential of radiomics features for predicting TP53 status preoperatively and guiding clinical treatment.

Radiomics Analysis of Computed Tomography for Prediction of Thyroid Capsule Invasion in Papillary Thyroid Carcinoma: A Multi-Classifier and Two-Center Study.

Objective: To investigate the application of computed tomography (CT)-based radiomics model for prediction of thyroid capsule invasion (TCI) in papillary thyroid carcinoma (PTC). Methods: This retrospective study recruited 412 consecutive PTC patients from two independent institutions and randomly assigned to training (n=265), internal test (n=114) and external test (n=33) cohorts. Radiomics features were extracted from non-contrast (NC) and artery phase (AP) CT scans. We also calculated delta radiomics features, which are defined as the absolute differences between the extracted radiomics features. One-way analysis of variance and least absolute shrinkage and selection operator were used to select optimal radiomics features. Then, six supervised machine learning radiomics models (k-nearest neighbor, logistic regression, decision tree, linear support vector machine [L-SVM], Gaussian-SVM, and polynomial-SVM) were constructed. Univariate was used to select clinicoradiological risk factors. Combined models including optimal radiomics features and clinicoradiological risk factors were constructed by these six classifiers. The prediction performance was evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA). Results: In the internal test cohort, the best combined model (L-SVM, AUC=0.820 [95% CI 0.758-0.888]) performed better than the best radiomics model (L-SVM, AUC = 0.733 [95% CI 0.654-0.812]) and the clinical model (AUC = 0.709 [95% CI 0.649-0.783]). Combined-L-SVM model combines 23 radiomics features and 1 clinicoradiological risk factor (CT-reported TCI). In the external test cohort, the AUC was 0.776 (0.625-0.904) in the combined-L-SVM model, showing that the model is stable. DCA demonstrated that the combined model was clinically useful. Conclusions: Our combined model based on machine learning incorporated with CT radiomics features and the clinicoradiological risk factor shows good predictive ability for TCI in PTC.

Laryngeal extra-skeletal Ewing sarcoma treated with DC-CTL immunotherapy: A case report and review of the literature.

Extra-skeletal Ewing sarcoma (EES) is a rare sarcoma composed primarily of small round cells, capable of metastasizing and relapsing. Few cases of EES originating from the larynx have been reported, and no publications regarding laryngeal EES treated with dendritic cells-cytotoxic T lymphocytes (DC-CTL) immunotherapy have been found. We described a 29-year-old woman with a mass found in the larynx. Diffuse small round cells with scanty cytoplasm shown by histology test and extremely positive staining of CD99 revealed by immunohistochemistry helped determine the diagnosis of laryngeal EES. The patient survived for seven years with no signs of recurrence or metastasis after six cycles of DC-CTL immunotherapy based on traditional treatments. This case indicates that DC-CTL immunotherapy could be considered a new option for treating EES.

Case Report: Two cases of survival after complete transection of the left common carotid artery.

Penetrating carotid artery traumas are rare yet fatal injuries with a high rate of mortality, and survivors may live with neurological sequelae. Of all the types of penetrating carotid artery traumas, the total transection of the common carotid artery (CCA) may be the most serious, can lead to death quickly, and has few reports of survivors. We described two cases of patients with complete CCA transections who survived without any neurological sequelae. The penetrating neck traumas of both patients were confirmed as complete CCA severance by CT and surgical exploration. Case 1 received the insertion of an interposition polytetrafluoroethylene graft to reconstruct the CCA, with postoperative ultrasound and CT angiography (CTA) verifying the total occlusion. Case 2 underwent nonoperative management under close observation and did not develop delayed active bleeding or neurological symptoms. Both patients recovered well, and no nervous system sequelae appeared during the follow-up period. A carotid artery injury cannot be ruled out in an asymptomatic penetrating neck injury. If CTA is feasible given the patient's hemodynamic condition, then it should be used as a routine examination to evaluate cervical vascular injury in patients with penetrating neck trauma. Management for hemodynamically stable carotid artery injuries remains controversial. These two cases of transverse carotid artery injury have caused us to further consider the principles of this kind of case management.

Computed Tomography-Based Radiomics Model to Predict Central Cervical Lymph Node Metastases in Papillary Thyroid Carcinoma: A Multicenter Study.

Objectives: This study aimed to develop a computed tomography (CT)-based radiomics model to predict central lymph node metastases (CLNM) preoperatively in patients with papillary thyroid carcinoma (PTC). Methods: In this retrospective study, 678 patients with PTC were enrolled from Yantai Yuhuangding Hot3spital (n=605) and the Affiliated Hospital of Binzhou Medical University (n=73) within August 2010 to December 2020. The patients were randomly divided into a training set (n=423), an internal test set (n=182), and an external test set (n=73). Radiomics features of each patient were extracted from preoperative plain scan and contrast-enhanced CT images (arterial and venous phases). One-way analysis of variance (ANOVA) and least absolute shrinkage and selection operator algorithm were used for feature selection. The K-nearest neighbor, logistics regression, decision tree, linear-support vector machine (linear-SVM), Gaussian-SVM, and polynomial-SVM algorithms were used to establish radiomics models for CLNM prediction. The clinical risk factors were selected by ANOVA and multivariate logistic regression. Incorporated with clinical risk factors, a combined radiomics model was established for the preoperative prediction of CLNM in patients with PTCs. The performance of the combined radiomics model was evaluated using the receiver operating characteristic (ROC) and calibration curves in the training and test sets. The clinical usefulness was evaluated through decision curve analysis (DCA). Results: A total of 4227 radiomic features were extracted from the CT images of each patient, and 14 non-zero coefficient features associated with CLNM were selected. Four clinical variables (sex, age, tumor diameter, and CT-reported lymph node status) were significantly associated with CLNM. Linear-SVM led to the best prediction model, which incorporated radiomic features and clinical risk factors. Areas under the ROC curves of 0.747 (95% confidence interval [CI] 0.706-0.782), 0.710 (95% CI 0.634-0.786), and 0.764 (95% CI 0.654-0.875) were obtained in the training, internal, and external test sets, respectively. The linear-SVM algorithm also showed better sensitivity (0.702 [95% CI 0.600-0.790] vs. 0.477 [95% CI 0.409-0.545]) and accuracy (0.670 [95% CI 0.600-0.738] vs. 0.642 [95% CI 0.569-0.712]) than an experienced radiologist in the internal test set in the combined radiomics model. The calibration plot reflected a favorable agreement between the actual and estimated probabilities of CLNM. The DCA indicated the clinical usefulness of the combined radiomics model. Conclusion: The combined radiomics model is a non-invasive preoperative tool that incorporates radiomic features and clinical risk factors to predict CLNM in patients with PTC.

Overexpression of TP53 mutation-associated microRNA-182 promotes tumor cell proliferation and migration in head and neck squamous cell carcinoma.

OBJECTIVE: To investigate the functional mechanism of microRNA-182 (miR-182) in head and neck squamous cell carcinoma (HNSCC). DESIGN: HNSCC and normal samples were used to check both p53 and miR-182 expression. Two cancer cell lines with overexpression of miR-182 were used to check cell proliferation and migration. In vivo role of miR-182 was assessed using mouse xenograft tumor model. ß-TrCP2 was found to be direct target of miR-182 by luciferase reporter assay. RESULTS: Overexpression of miR-182 was closely related with overproduction of p53 in HNSCC. Overexpression of miR-182 promoted cell proliferation and migration and its oncogenic effect was also confirmed in vivo. ß-TrCP2 acted as direct target of miR-182 and its overexpression can reverse the miR-182 induced cell proliferation and migration. CONCLUSION: Overexpression of TP53 mutation-associated miR-182 may promote tumor cell proliferation and migration in HNSCC and suggest possible biomarker for the prediction of tumor recurrence.

Analysis of Association Between MGMT and p53 Gene Single Nucleotide Polymorphisms and Laryngeal Cancer.

AIM: To investigate the p53 and O6-methylguanine DNA methyltransferase (MGMT)5' upstream sequence gene promoter regions for single nucleotide polymorphisms and explore the p53 gene 5' upstream sequence consisting of two haplotypes to provide a genetic marker for the incidence of laryngeal squamous cell carcinoma. MATERIALS AND METHODS: We included 96 cases of laryngeal squamous cell carcinoma and 102 controls. We used SNaPshot micro-sequencing analysis of the MGMT promoter region for four single nucleotide polymorphisms and p53 gene 5' upstream sequence loci (rs1625649, rs2287499, rs2287498, rs228749) genotypes. We calculated and compared two groups for genotypic and allelic frequencies, applied HaploView4.2 for computing rs2287499, rs2287498, rs228749 values and haplotype frequencies and tested control loci and Hardy-Weinberg equilibrium. All the experimental data were statistically evaluated using SPSS17.0. The Chi-square test was used for statistical analysis with p<0.05 indicating statistical significance. RESULTS: 5'Upstream single nucleotide polymorphisms rs1625649, rs2287499, rs2287498, rs228749 of p53 were polymorphic in both patient and control groups. There was no statistical significance in frequency distributions for the four loci genotypes when comparing patients and healthy controls (Chi-square values were 4.47, 0.98, 1.67, 4.68, respectively; p>0.05). However, allelic frequencies of the MGMT promoter region locus rs1625649 between patients and healthy control groups were statistically significantly different (chi-square value of 5.77; p<0.05). Differences between allelic frequencies for the p53 gene 5' upstream sequence loci rs2287499, rs2287498 and rs228749 between patients and the healthy control group were not statistically significant (Chi-square values were 1.11,1.56,3.36; p>0.05). Nor were those for the two haplotypes of rs2287499, rs2287498 and rs228749 between patients and the healthy control group were not statistically significant (Chi-square value 1.46, p>0.05). CONCLUSION: MGMT gene polymorphism appears to be associated with the incidence of laryngeal cancer.

Application of Ultrasound-guided Core Needle Biopsy in the Diagnosis of T3 or T4 Stage Laryngeal and Hypopharyngeal Cancer.

AIM: To study the value of ultrasound-guided core needle biopsy (CNB) in the diagnosis of T3 or T4 stage laryngeal and hypopharyngeal cancer, which is difficult by routine methods. PATIENTS AND METHODS: Nineteen cases of T3 or T4 stage laryngeal or hypopharyngeal carcinoma with abnormal pharyngeal sensitivity, severe dyspnea, submucous cancer recurrence, cardiovascular and pulmonary dysfunction were reviewed retrospectively from October 2012 to October 2014 in the Yuhuangding Hospital of Qingdao University. Ultrasound-guided coarse needle biopsies were used on primary lesions after assessing the patients with neck-enhanced computed tomography (CT) or magnetic resonance imaging (MRI) scan(s). The clinical value of ultrasound-guided CNB in the diagnosis of laryngeal and hypopharyngeal cancer was analyzed. RESULTS: All patients underwent successful pathological diagnosis by ultrasound-guided CNB without any serious complications. Dyspnea, cardiovascular and pulmonary dysfunction did not deteriorate. CONCLUSION: Ultrasound-guided CNB is a highly safe and efficient method for the pathological diagnosis of T3 or T4 stage laryngeal and hypopharyngeal cancer. It should be used especially when the fiberoptic or laryngoscope biopsy are of high risk.

[The diagnosis and treatment of one huge cystic lymphangioma in etropharyngeal space].

One child was hospitalized because of repeated cough and sputum. The biopsy diagnosis in local hospital was cystic lymphangioma in retropharyngeal space. We carried out transoral incision and drainage by catheter under general anesthesia. Put into the surgical cavity a suction drainage tube, and injected 5 mg dexamethasone and 8mg Bleomycin. He had nasogastric liquid diet after operation. We removed the suction drainage tube two weeks later. No recurrence was found following up over two years.

MGMT Leu84Phe polymorphism contributes to cancer susceptibility: evidence from 44 case-control studies.

BACKGROUND: O(6)-methylguanine-DNA methyltransferase is one of the few proteins to directly remove alkylating agents in the human DNA direct reversal repair pathway. A large number of case-control studies have been conducted to explore the association between MGMT Leu84Phe polymorphism and cancer risk. However, the results were not consistent. METHODS: We carried out a meta-analysis of 44 case-control studies to clarify the association between the Leu84Phe polymorphism and cancer risk. RESULTS: Overall, significant association of the T allele with cancer susceptibility was verified with meta-analysis under a recessive genetic model (P<0.001, OR=1.30, 95%CI 1.24-1.50) and TT versus CC comparison (P=0.001, OR=1.29, 95% CI 1.12-1.50). In subgroup analysis, a significant increased risk was found for lung cancer (TT versus CC, P=0.027, OR=1.67, 95% CI 1.06-2.63; recessive genetic model, P=0.32, OR=1.64, 95% CI 1.04-2.58), whereas risk of colorectal cancer was significantly low under a dominant genetic model (P=0.019, OR=0.84, 95% CI 0.72-0.97). Additionally, a significant association between TT genetic model and total cancer risk was found in the Caucasian population (TT versus CC, P=0.014, OR=1.29, 95% CI 1.05-1.59; recessive genetic model, P=0.009, OR=1.31, 95% CI 1.07-1.61), but not in the Asian population. An increased risk for lung cancer was also verified in the Caucasian population (TT versus CC, P=0.035, OR=1.62, 95% CI 1.04-2.53; recessive genetic model, P=0.048, OR=1.57, 95% CI 1.01-2.45). CONCLUSIONS: These results suggest that MGMT Leu84Phe polymorphism might contribute to the susceptibility of certain cancers.