Lentivirus-Mediated Short Hairpin RNA for Follistatin Downregulation Suppresses Tumor Progression in Hypopharyngeal Carcinoma

Objective:Follistatin(FST)inhibits the action of activin by interfering with the binding of activin to its receptor.Although the prognostic value of FST in various cancers has been investigated previously,studies rarely focused on hypopharyngeal carcinoma(HPC).In our study,the effect of FST expression on HPC tissues and cell lines was investigated.Methods:A total of 60 patients with HPC were recruited for this study.Levels of FST mRNA and protein were measured by quantitative polymerase chain reaction(PCR)and immunohistochemistry in HPC tissue samples and by qPCR in the HPC FaDu cells,as well as immortal nasopharyngeal epithelial cell line NP-69 cells.After silencing the FST expression in FaDu cells using lentivirus-mediated siRNA that was specific for FST mRNA,cell proliferation was determined by a Celigo assay.Tumor growth was monitored in nude mice and viability was determined by a methylthiazoletetrazolium assay.The ratio of cell cycle arrest and apoptosis was evaluated by flow cytometry.The colony formation ability was performed using Giemsa staining.In addition,wound healing and Transwell migration and invasion assays were performed for the analysis of cell motility.Results:FST expression was significantly higher in human HPC tissue and FaDu cells than in normal tissue and NP-69 cells.A higher expression of FST in HPC samples was positively correlated with advanced tumors.Moreover,FST knockdown by shRNA significantly decreased cell growth,colony formation,migration and invasion.Furthermore,FST silencing increased the cell apoptosis percentage,and arrested cell cycle in the S phase in FaDu cells.In addition,FST silencing suppressed tumor growth in vivo.Conclusions:Our results indicated that the FST gene was associated with HPC progression and may serve as a potential therapeutic target for the treatment of HPC.

The Value of Magnetic Resonance Diffusion-Weighted Imaging in Evaluating the Efficacy of Concurrent Chemoradiotherapy in Patients with Hypopharyngeal Carcinoma

Objective: To investigate the application value of magnetic resonance diffusion-weighted imaging (DWI) combined with conventional magnetic resonance imaging (MRI) in evaluating the efficacy of concurrent chemoradiotherapy for hypopharyngeal carcinoma. Methods: A total of 20 patients with hypopharyngeal carcinoma diagnosed by pathological biopsy (who only received chemoradiotherapy without surgery) were collected. Before treatment, all patients underwent conventional MRI and DWI scanning, MRI characteristics of patients were analyzed, and maximum cross-sectional area of the tumor and average apparent diffusion coefficient (ADC) value were measured. One month after treatment, MRI was performed again to measure residual tumor area and ADC value, and the tumor remission rate was calculated. The changes in tumor ADC values before and after treatment were analyzed and their correlation with tumor remission rate was analyzed. The differences in ADC values and changes between complete response patients (CR group) and incomplete response patients (non-CR group) before and after treatment were analyzed. Results: The tumor area of 20 patients with hypopharyngeal cancer was 3.48 (0.93 - 5.6) cm2 before treatment and 0.24 (0 - 0.9) cm2 after treatment. There were 15 patients (15/20, 75%) in the CR group and 5 patients (5/20, 25.0%) in the non-CR group. The remission rate was 90.3% (6.0% - 100%). The average ADC value of the tumor before treatment was negatively correlated with the tumor remission rate after treatment (r = ?0.786, ?0.813, P Conclusion: The ADC value of tumor before treatment and the change of ADC value of tumor before and after treatment have a certain significance in evaluating the early remission rate of hypopharyngeal carcinoma after chemoradiotherapy.

HSP90AA1 promotes lymphatic metastasis of hypopharyngeal squamous cell carcinoma by regulating epithelial-mesenchymal transition

Lymphatic metastasis(LM)emerges as an independent prognostic marker for hypopharyngeal squamous cell carcinoma(HSPSCC),chiefly contributing to treatment inefficacy.This study aimed to scrutinize the prognostic relevance of HSP90AA1 and its potential regulatory mechanism of concerning LM in HPSCC.Methods:In a preceding investigation,HSP90AA1,a differential gene,was discovered through transcriptome sequencing of HPSCC tissues,considering both the presence and absence of LM.Validation of HSP90AA1 expression was accomplished via qRT-PCR,western-blotting(WB),and immunohistochemistry(IHC),while its prognostic significance was assessed employing Kaplan–Meier survival analysis(KMSA),log-rank test(LR),and Cox’s regression analysis(CRA).Bioinformatics techniques facilitated the prediction and analysis of its plausible mechanisms in LM,further substantiated by in vitro and in vivo experiments utilizing FaDu cell lines.Results:HSP90AA1 is substantially upregulated in HPSCC with LM and is identified as an independent prognostic risk determinant.The down-regulation of HSP90AA1 can achieve inhibition of tumor cell proliferation,migration and invasion.Both in vivo experiments and Bioinformatics exploration hint at promoting LM by Epithelial-mesenchymal transition(EMT),regulated by HSP90AA1.Conclusions:HSP90AA1,by controlling EMT,can foster LM in HPSCC.This finding sets the foundation for delving into new therapeutic targets for HPSCC.

Development of a Nomogram Based on Clinicopathological and Biological Features to Predict Neck Lymph Node Metastasis in Hypopharyngeal Squamous Cell Carcinoma

Background: Clinicopathological and biological features are associated with neck lymph node metastasis (LNM) of hypopharyngeal squamous cell carcinoma (HSCC). However, there is no complete nomogram combining multiple factors that can be used to accurately predict the neck LNM status for HSCC patients. Purpose: To guide the selection of surgical methods and radiotherapy areas for hypopharyngeal cancer. In this study, a nomogram was developed to combine these risk factors to predict neck LNM and guide the treatment of HSCC. Material and Methods: This retrospective study included 117 patients (training cohort, 64 patients;trial cohort, 53 patients). Biological characteristics of HSCC patients were assessed using immunohistochemical staining, and data of patient age, gender, and preoperative computed tomography (CT) scan reports were collected. Significant risk factors in univariate analysis were further identified to be independent variables in multivariate logistic regression analysis, which were then incorporated in and presented with a nomogram by using the rms package in R software. Receiver operating characteristic (ROC) curves and calibration curves were used to validate the discrimination and accuracy in the training and validation cohorts, respectively, and clinical usefulness was verified in decision curve analysis curves. Results: All variables with P-values < 0.2 in the univariate analysis were selected for multivariate logistic regression analysis to further identify independent risk factors for neck LNM. In multivariate logistic regression analysis, variables with P-values < 0.2 were identified as independent risk factors and then used to construct the nomogram. In total, five independent predictors, including the maximum tumor diameter in CT, tumor cell differentiation, LNM status in CT, Stathmin1 expression level, and lymphatic vessel invasion were included in the nomogram. The area under the ROC curve (AUC) was 0.916 (95% confidence interval [CI], 0.833 - 1.000) and AUC of 0.928 (95% CI, 0.864 - 1.000) in internal validation and the external validation. Conclusions: Both the internal validation in the training cohort and the external validation in the validation cohort showed that the nomogram had good discrimination, accuracy, and excellent clinical usefulness. The nomogram based on clinicopathological and biological features developed in this study has strong predictive power and could be used to predict neck LNM of HSCC in clinical practice.

Positron emission tomography and magnetic resonance imaging combined with computed tomography in tumor volume delineation: A case report

BACKGROUND Accurate delineation of the target area for patients with hypopharyngeal cancer is the key to achieving an ideal radiotherapy effect.Since computed tomography(CT)alone can no longer meet the treatment needs,fusing CT images with magnetic resonance imaging(MRI)or positron emission tomography(PET)images can overcome the disadvantages of CT.Herein,we present a clinical case of hypopharyngeal cancer to delineate the tumor volume using combined MRI-CT and PET-CT fusion images to examine if they could accurately cover the tumor volume.CASE SUMMARY A 67-year-old male patient with hypopharyngeal carcinoma could not tolerate chemotherapy and surgery due to complicated health issues such as diabetic nephropathy and other underlying diseases.After multidisciplinary consultations,clinicians eventually agreed to undergo radiotherapy to control the progression of his tumor.He was examined by CT,MRI,and 18-fluorodeoxyglucose-PET for treatment planning,and CT images were fused with PET and MRI images while delineating tumor volume.CONCLUSION The image fusion of MRI-CT and PET-CT has both advantages and disadvantages.Compared with CT images alone,the combination of MRI-CT and PET-CT fusion images can precisely cover the gross tumor volume in hypopharyngeal carcinoma and avoid overestimation or incomplete coverage of tumor volume.