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Purpose: To investigate the potential mechanisms of miR-211-3p on induction chemotherapy (IC) sensitivity in hypopharyngeal squamous cell carcinoma (HSCC)., Methods: qRT-PCR was assessed to compare the miR-211-3p expression between IC sensitive and insensitive tumor tissues. The MTT assay was performed to analyze cell proliferation and viability to paclitaxel after alteration of miR-211-3p. Flow cytometry assay was conducted to explore cell apoptosis. Transwell assay was used to explore the effect of miR-211-3p on cell migration. Transcriptome sequencing was then performed to select differentially expressed genes (DEGs) after over-expression of miR-211-3p. GO and KEGG enrichment analyses were conducted to annotate DEGs. PPI analysis was conducted to screen candidate genes. The differential expression and survival status of candidate genes were further validated in TCGA-HNSCC data. The single sample GSEA method was used to investigate the association between downstream genes and immune cell infiltration., Results: miR-211-3p was up-regulated in IC insensitive larynx-hypopharyngeal tumor tissues. Over-expression of miR-211-3p promoted cell proliferation and migration, and inhibited apoptosis. The IC 50 value of miR-211-3p overexpression (OE) group was significantly higher than negative control (NC) group treated with paclitaxel, suggesting miR-211-3p enhanced IC insensitivity in HSCC. We found 778 DEG after over-expression of miR-211-3p and 11 significant genes were then identified. Finally, colony stimulating factor 2 (CSF2) and C-C motif chemokine ligand 20 (CCL20) were validated to be significantly high expressed and associated with poorer overall survival in head and neck squamous cell carcinoma, which were involved in TNF signaling pathway and then regulated immune cell infiltration., Conclusion: The miR-211-3p could promote HSCC progression and upregulate CSF2/CCL20/TNF signaling to promote IC insensitivity in HSCC, which may provide new ideas for HSCC therapy., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Overexpression of long non-coding RNAs (lncRNAs) reveals the abnormal pathological processes in many human cancers. KRT16P3, a novel overexpressed lncRNA in tongue squamous cell carcinoma (TSCC), was identified by previous lncRNA microarrays. However, the role of KRT16P3 in TSCC is not clear., Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of KRT16P3 in TSCC tissues and cells. Next, the relationships between KRT16P3 and the clinical significance of TSCC patients were analyzed. Additionally, Cell Counting Kit-8, 5-Bromo-2-deoxyuridine (BrdU) incorporation assay, cell colony formation assay, flow cytometry cell apoptosis analysis, scratch wound healing assay, transwell invasion assay were used to explore the biological function of KRT16P3. Western blot and qRT-PCR were used to determine the expression of epithelial-mesenchymal transition (EMT) markers. The pathway changes after KRT16P3 knockdown were detected by Western blot., Results: We found KRT16P3 expression is significantly upregulated in TSCC tissues and positively associated with advanced clinicopathological features of TSCC patients, and it may serve as a poor prognostic factor. Functionally, KRT16P3 knockdown inhibits proliferation, migration, invasion and promotes apoptosis of TSCC cells. Furthermore, we also revealed that KRT16P3 knockdown suppresses EMT and JAK2/STAT3 signaling pathway., Conclusion: Our results validated that KRT16P3 can modulate the malignant progression, EMT process, and JAK2/STAT3 signaling pathway of TSCC, which might also serve as a novel prognostic biomarker and an attractive target for TSCC patients., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: The prognostic factors and survival difference between inverted papilloma (IP)-associated sinonasal squamous cell carcinoma (SCC) and de novo SCC are unclear., Objective: This study aimed to compare the clinical features and oncologic outcomes in patients with IP-associated SCC and de novo SCC; and additionally, to analyze the prognostic factors of the two types of SCCs., Methods: Data from 173 SCC patients treated for IP-associated SCC (n = 89) and de novo SCC (n = 84), were reviewed retrospectively for demographic features, tumor characteristics, treatment modality, and clinical outcomes. 5-year overall survival (OS) and disease free survival (DFS) was analyzed using the Kaplan-Meier method, and Cox proportional hazards model was used to analyze factors influencing prognosis., Results: A higher proportion of IP-associated SCC occurred in frontal and sphenoid sinus compared to de novo SCC. The two groups demonstrated similar 5-year OS and DFS (5-year OS: 63.3% and 55.4%, DFS: 45.4% and 50.1%, respectively). The metachronous tumor had a relatively better prognosis outcome than synchronous tumor and de novo SCC (5-year OS: 73.1%, 54.5% and 55.4%, respectively). Both groups showed similar loco-regional recurrence rates ( p  > 0.05); however, de novo SCC tumors demonstrated an increased incidence of distant metastasis. Multivariate analysis indicated that age >70 years, advanced tumor stage and surgical margin were independent predictive factors for the risk of mortality (HR 2.047, 1.581 and 1.931, respectively)., Conclusion: IP-associated SCCs have an aggressive loco-regional tendency, whereas de novo SCCs have a higher aggressive distant metastatic propensity. Age, tumor stage and surgical positive margin are key factors for poor prognosis and should be routinely taken into consideration during treatment planning and subsequent surveillance.

Objective: To evaluate the value of iodine staining in the margin determination in transoral laser microsurgery for early glottic carcinoma. Method: 35 patients with early glottic laryngeal cancer（Tis-T2） were enrolled in this study from January 2016 to December 2017.All patients were applied with 1.5% Lugol iodine solution to vocal cords for 3 times after general anesthesia，and the coloration of the laryngeal cavity was observed under the operation microscope to determine the lesion margins.The lesions were excised completely with CO₂ laser.The complications，complete resection rate and recurrence rate were evaluated. Result: In all patients，iodine staining was used to assist in determining the lesion range during transoral laser microsurgery.None of the patients had complications.Postoperative pathological results showed that the complete resection rate was 88.6%.There were no local recurrence cases in Tis and T1a，while local recurrence rate was 12.5% in T1b and 16.7% in T2，respectively.The overall local recurrence rate was 5.7%. Conclusion: The application of iodine staining in transoral laser microsurgery for early glottic carcinoma may help improve the complete resection rate and local control rate.It is a reliable，simple and low-cost method，worthy of further clinical application.Key words: laryngeal neoplasms;glottis; transoral laser microsurgery; iodine staining., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Background: This study aimed to analyze the prognostic factors and survival outcomes of squamous cell carcinoma (SCC) originating from sinonasal inverted papillomas (IPs), based on data from a single institution., Methods: The data from 120 patients treated at the affiliated Beijing Tongren Hospital, Capital Medical University, for SCC originating from sinonasal IPs between 2005 and 2018 were retrospectively reviewed. Data related to demographic features, tumor characteristics, treatment modality, and clinical outcomes were collected. Survival data were investigated using the Kaplan-Meier method, Cox regression analysis, and the nomogram model predictive of survival probabilities., Results: Among 1034 patients with sinonasal IPs, 120 patients (11.6%) with malignancy were identified. The overall survival (OS) and disease-free survival (DFS) rates at 5 years were 56.0% and 42.3%, respectively. Multivariate analysis showed that synchronous tumors and tumor stage were independent predictive factors for the risk of mortality (hazard ratio [HR], 1.954; 95% confidence interval [CI], 1.022-3.737, p = 0.043; HR, 1.737, 95% CI, 1.095-2.770, p = 0.020, respectively). The surgical margin was another important independent predictor, with patients with negative margins demonstrating a more than 2-fold improved survival compared to those with positive margins (HR, 2.095; 95% CI, 1.031-4.243; p = 0.041)., Conclusion: The main factors affecting the prognosis and outcomes were synchronous tumors, advanced tumor stage, and positive surgical margins. These findings highlight the importance of tumor biology and early detection in patient outcomes. In addition, risk factors should be taken into consideration during treatment planning and subsequent tumor surveillance., (© 2019 ARS-AAOA, LLC.)

Rationale: posterior pharyngeal wall carcinomas located at the level of the cricoid cartilage is a rare occurrence. Surgical treatment is a dilemma owing to the retention of laryngeal function PATIENT CONCERNS:: A 56-year-old man underwent a "resection of left cervical tumor" at another hospital. Postoperative pathological report noted a left metastatic squamous cell carcinoma of the neck. Neck-enhanced computed tomography (CT) revealed thickening of the right posterior pharyngeal wall, and an image of soft tissue density was present on the posterior pharyngeal wall with a size of approximately 1.0 × 1.5 cm., Diagnosis: Hypopharyngeal carcinoma (T1N2M0)., Interventions: We performed operation on a patient who suffered from posterior pharyngeal wall carcinomas at the level of the cricoid cartilage. The postoperative defects were repaired with posterior pharyngeal flaps., Outcomes: The patient with posterior pharyngeal flap after operation survived without necrosis, the neck wounds of patient healed in 1 stage, and the phonation, swallowing and respiratory function was good., Lessons: The application of posterior pharyngeal flap to repair defects due to posterior pharyngeal wall carcinomas that were located at the level of the cricoid cartilage after resection is an effective reconstruction method that is easy to use and has excellent repair effects.

Purpose: To identify potential molecular markers for induction chemotherapy of Laryngeal squamous cell carcinoma (LSCC)., Methods: Differently expressed genes between chemo-sensitive group (seven cases) and chemo-insensitive (five cases) group after induction chemotherapy by TPF were identified by microarrays. Bayes network and Random forest analyses were employed to identify core genes for induction chemotherapy. The diagnostic value of these core genes was also evaluated by ROC analysis., Results: Six genes (SPP1, FOLR3, KYNU, LOC653219, ADH7 and XAGE1A) are highly expressed, while seven gene (CADM1, NDUFA4L2, CCND2, RARRES3, ERAP2, LYD6 and CNTNAP2) present significantly low expression. Among these genes, genes CADM1, FOLR3, KYNU, and CNTNAP2 are core candidates for LSCC chemo-sensitivity. And that the low expression of CADM1 may result in chemo-sensitivity, which leads to high expression of gene FOLR3 and KYNU, and low expression of gene CNTNAP2. Besides, ROC analysis shows that these four genes exhibit effective diagnostic value for induction chemo-sensitivity., Conclusions: CADM1 may be a potential molecular marker for LSCC induction chemotherapy, while CADM1, FOLR3, KYNU, and CNTNAP2 may provide essential guidance for LSCC diagnosis and follow-up treatment strategies.

Background: Narrow band imaging (NBI) and stroboscopy are non-invasive techniques to detect the malignant lesions of the vocal cord. This study was to assess the diagnostic value of combined endoscopic analysis in the applanate indiscernible early-stage vocal cord cancer., Methods: A total of 110 patients with 160 suspicious vocal cord malignant lesions were included in this retrospective study. Stroboscopy was immediately performed after NBI and white light endoscopy (WLE) were performed in all patients. Excisional biopsy was performed to examine histopathology examination., Results: We found that the diagnostic specificity and PPV were higher in the NBI and WLE combined with stroboscopy group than in the NBI and WLE group without stroboscopy (88.9% vs 72.5%, 88.4% vs 60.9%). However, the diagnostic sensitivity was not significantly different in those two groups (69.3% vs 67.7%)., Conclusion: NBI and WLE combined with stroboscopy is a promising method to detect early-stage vocal cord cancer with the advantage of clinical feasibility and diagnostic specificity.

Laryngeal squamous cell carcinoma (LSCC) is one of the most common types of head and neck malignant tumor; however, there is a lack of effective molecular targets for therapy. The present study detected the expression of three immunity‑associated molecules [forkhead box (FOX)3, interleukin (IL)‑10 and cluster of differentiation (CD)274] in 133 LSCC samples using immunohistochemistry (IHC); subsequently, the association between their expression and the clinical characteristics of LSCC were analyzed. Spearman's rank correlation method, Kaplan‑Meier and Cox regression model were used to analyze the correlations of the three proteins and their clinical significance. StarBase and miRTarBase databases were used to establish the competitive endogenous (ce)RNA network of the three molecules. IHC demonstrated that the positive expression rates of FOXP3, IL‑10 and CD274 were 68.4, 73.7 and 58.6% in 133 LSCC samples, respectively. In addition, it was identified that the expression of the three proteins was closely correlated with the clinical characteristics of LSCC, including lymph node metastasis and prognosis (P<0.05). There was also a significant association of co‑expression between any two proteins (P<0.001). Furthermore, the expression levels of FOXP3, IL‑10 and CD274 were negatively associated with the survival rate of patients with LSCC (P<0.05). The results of a Cox regression model suggested that the three proteins were prognostic risk factors for LSCC (P<0.05). The ceRNA network revealed that 10 microRNAs (miRs; including miR‑16‑5p and miR‑214‑3p), 123 long non‑coding RNAs (including X‑inactive specific transcript, H19 and metastasis associated lung adenocarcinoma transcript 1) and 408 circular RNAs (including ATP‑binding cassette subfamily C member 1 hsa&#95;circ&#95;001569 and ISY1 splicing factor homolog hsa&#95;circ&#95;001859) may regulate the expression of FOXP3, IL‑10 and CD274. The data generated from the present study may increase the understanding of the immune escape mechanisms of LSCC and may be beneficial for the development of a specific immunotherapy.

Solute carrier family 7, membrane 11 (SLC7A11) or (xCT) is a component of the cysteine-glutamate transporter, which plays a critical role in glutathione homeostasis which is important to protect cells from oxidative stress. SLC7A11 is distributed in various tissues and participates in the occurrence of a number of diseases, particularly in the pathogenesis of malignant tumors, but its role in laryngeal cancer development has not yet been clearly defined. The objective of the present study was to investigate the role of SLC7A11 in laryngeal squamous cell carcinoma (LSCC). We conducted immunohistochemistry and RT-PCR to evaluate the protein and mRNA levels of SLC7A11 in LSCC and in control tissues, respectively. The knockdown experiments were conducted with SLC7A11 short hairpin RNA (shRNA) lentivirus, and the protein and mRNA levels of SLC7A11 were assessed by RT-PCR and western blotting. The functional study of SLC7A11 in vitro was conducted by MTT assay, and the effects on the cell cycle were detected using flow cytometry. Immunohistochemical results revealed that the expression levels of SLC7A11, Ki-67 and p53 in LSCC tissues were higher than those in laryngeal dysplasia tissues. The Spearman rank correlation analysis revealed that the expression of SLC7A11 was positively correlated with the expression of p53 and Ki-67. Cox regression analysis and Kaplan-Meier plots confirmed that the expression levels of SLC7A11 were a prognostic factor for overall survival (OS) rates and postoperative recurrence of LSCC. Moreover, the functional study of SLC7A11 in vitro revealed that knockdown of SLC7A11 using shRNA inhibited cell proliferation by inducing cell cycle arrest at the G1 phase. Immunohistochemical and RT-PCR results and knockdown experiments of SLC7A11 revealed that SLC7A11 was involved in the progression of LSCC, and may provide clinical information for the evaluation of OS rates and postoperative recurrence of LSCC. Collectively, these observations suggest that SLC7A11 may be a vital biomarker for the diagnosis and prognosis in human LSCC, and targeting SLC7A11 appears to be a potentially significant method for LSCC treatment.

Purpose: The minichromosomal maintenance (MCM) proteins are involved in the initiation and DNA replication. The role of MCM4 remains to be elucidated. The purpose of this study was to investigate the effects of MCM4 in laryngeal squamous cell carcinoma (LSCC) cell growth and apoptosis., Methods: LSCC cell line UMSCC 5 was used in this study. The small interfering RNA (siRNA) of MCM 4 gene was used to identify the effects of MCM4 on the proliferation and apoptosis using methylimidazole tetrazolium (MTT) assay and flow-cytometry, respectively. Confirmed LSCC and adjacent non-tumor tissues were collected from 34 patients who were willing to participate in the study, from 2010 through 2015, from 163 patients undergoing treatment in the Department of Otorhinolaryngology/Head and Neck Surgery of Beijing Tongren Hospital in Capital Medical University of P.R. China. Immunohistochemical staining of MCM4 expression in the resected tissues was performed to analyze the correlation between its expression and the clinicopathological characteristics., Results: The results showed that siRNA of MCM4 could significantly inhibit LSCC cell line UMSCC 5 proliferation and induce apoptosis. MCM4 mRNA was higher expressed in carcinoma tissues than in adjacent normal tissues. MCM4 expression was correlated with male gender, smoking history and poor differentiation., Conclusions: We noticed a significant role for MCM4 overexpression in human LSCC tissues and their corresponding adjacent non-neoplastic tissues and found that siRNA of MCM4 can significantly decrease the proliferation of cancer cells. It is suggested that MCM4 profiling could potentially be used to predict response to treatment and prognosis in LSCC.

Conclusion: These results provided a battery of genes relating to TPF chemotherapeutic sensitivity and might act as molecular targets in laryngeal squamous cell carcinoma (LSCC) treatment. Moreover, these candidate biomarkers could contribute to LSCC individualized treatment., Objectives: To screen out a set of candidate genes which could help to determine whether patients with LSCC could benefit from TPF induction chemotherapy., Method: Gene-expression profiles in seven TPF-sensitive patients were compared to four resistant controls by microarray analysis. Subsequently, expression levels of potential biomarkers in chemosensitive cell line UMSCC5 after TPF treatment were observed by qRT-PCR., Results: Through microarray analysis, 1546 differently expressed genes were identified, of which 769 were up-regulated in TPF chemotherapy-responsive tissues, whereas 777 were down-regulated. Gene ontology (GO) analysis suggested these genes participating in physiological processes including cell differentiation, metabolism, signal transduction, and cellular component organization. Additionally, Kyoto Encyclopedia of Genes and Genomes (KEGG) database revealed that Wnt and p53 signaling pathways occupied important roles in TPF chemotherapeutic sensitivity. Moreover, in vitro cell culture experiments revealed the expression alternations of Mapk10, Jun, Vegfb, Pik3r5, Pld1, Tek, Itga6 exposed to TPF treatment by qRT-PCR, whilst providing an insight into the mechanism underlying TPF chemotherapeutic response in LSCC.

Long non-coding RNA (lncRNA) plays an important role in tumorigenesis. However, the expression pattern and function of lncRNAs in laryngeal squamous cell carcinoma (LSCC) are still unclear. To investigate the aberrantly expressed lncRNAs and mRNAs in advanced LSCC, we screened lncRNA and mRNA expression profiles in 9 pairs of primary Stage IVA LSCC tissues and adjacent non-neoplastic tissues by lncRNA and mRNA integrated microarrays. Gene Ontology and pathway analysis were performed to find out the significant function and pathway of the differentially expressed mRNAs, gene-gene functional interaction network and ceRNA network were constructed to select core mRNAs, and lncRNA-mRNA expression correlation network was built to identify the interactions between lncRNA and mRNA. qRT-PCR was performed to further validate the expressions of selected lncRNAs and mRNAs in advanced LSCC. We found 1459 differentially expressed lncRNAs and 2381 differentially expressed mRNAs, including 846 up-regulated lncRNAs and 613 down-regulated lncRNAs, 1542 up-regulated mRNAs and 839 down-regulated mRNAs. The mRNAs ITGB1, HIF1A, and DDIT4 were selected as core mRNAs, which are mainly involved in biological processes, such as matrix organization, cell cycle, adhesion, and metabolic pathway. LncRNA-mRNA expression correlation network showed LncRNA NR&#95;027340, MIR31HG were positively correlated with ITGB1, HIF1A respectively. LncRNA SOX2-OT was negatively correlated with DDIT4. qRT-PCR further validated the expression of these lncRNAs and mRNAs. The work provides convincing evidence that the identified lncRNAs and mRNAs are potential biomarkers in advanced LSCC for further future studies., Competing Interests: The authors have declared that no competing interests exist.

Tumor necrosis factor superfamily member 13 (TNFSF13) modulates cell proliferation and apoptosis and participates in the pathogenesis of solid tumors, but its role in laryngeal cancer development is not clearly defined. In order to investigate whether TNFSF13 can be used as a biomarker for diagnosis and prognosis in laryngeal squamous cell carcinoma (LSCC) and the role of TNFSF13 in laryngeal cancer carcinogenesis, we conducted immunohistochemistry and ELISA assays to evaluate the expression level of TNFSF13 in laryngeal cancer patients and the contrast. We also conducted experiments on the functional study of TNFSF13 in vitro. We found that the expression levels of TNFSF13, ki-67, and NF-κB p65 in LSCC tumor tissues were higher than those in vocal polyp and para-carcinoma tissues. The Spearman rank correlation analysis showed that the expression of TNFSF13 had a positive correlation with the expression of ki-67 and NF-κB p65. Cox regression analysis and Kaplan-Meier plots confirmed the expression level of TNFSF13 was a prognostic factor for LSCC. Moreover, the serum TNFSF13 level was significantly higher in LSCC patients than in the controls, and the serum expression level of TNFSF13 can distinguish LSCC from healthy people, precancerosis, or laryngeal benign tumor. In addition, functional study of TNFSF13 in vitro revealed that knockdown of TNFSF13 inhibited cell proliferation by inducing G1 phase cell cycle arrest in Hep-2 cells. In conclusion, TNFSF13 may be a potential novel molecular target for diagnosis and prognosis in human LSCC, and therapies that target TNFSF13 may have clinical significance for the treatment of LSCC.

Zinc finger protein, X-linked (ZFX) gene locus on the human X chromosome is structurally similar to the zinc finger protein, Y-linked gene. Its role in human laryngeal squamous cell carcinoma (LSCC) is still not clearly defined. This study was focused on investigating the role of zinc-finger protein X-linked (ZFX) in human LSCC. Expression levels of ZFX were examined in LSCC tissues, corresponding adjacent non-tumoral tissues and vocal leukoplakia tissues by immunohistochemistry (IHC). The association with the expression level of ZFX and LSCC clincopathological parameters was analyzed. The prognostic value of ZFX expression was also analyzed. Lentivirus-mediated RNA interference was applied to silence ZFX expression and the effects of ZFX knockdown on the growth of human LSCC primary cells was investigated. Overexpression of ZFX was found in LSCC tissues. The expression of ZFX was associated with the clinical stage of LSCC. Patients with higher level of ZFX experienced a poorer prognosis compared to those with lower level of ZFX. Knockdown of ZFX inhibited cell proliferation, colony formation and migration of LSCC primary cells. Moreover, ZFX silencing induced cell apoptosis. These results provide the convincing evidence for the first time that ZFX plays an important role in LSCC development and could be a potential therapeutic target or prognostic predictor for LSCC.

Zinc finger protein, X-linked (ZFX) is a transcriptional factor involved in many physiological processes such as embryonic stem cell survival and self-renewal. Though ZFX dysfunctions have been identified in variant human diseases and especially in cancers, its pathological roles have not been fully addressed. Here, we explored the relationship between ZFX expression and squamous cell carcinoma (SCC) of the tongue. We found that ZFX expression was significantly higher in tongue SCC tumors as compared to tumor-adjacent normal tissues. Furthermore, ZFX knockdown impeded cell proliferation, impaired colony formation ability, and lead to cell cycle arrest while induced cell apoptosis in human tongue squamous cell carcinoma cell line Tca-8113. Our results provide evidence suggesting that ZFX overexpression is associated with the development of tongue SCC and ZFX knockdown is a potential treatment for tumor suppression.

MicroRNAs (MiRNAs) have been recognized to regulate cancer initiation and progression in carcinogenesis as either oncogenes or tumor suppressor genes, but their role in hypopharyngeal cancer development is not clearly defined. To determine whether miRNA-203 can promote tumor growth in human hypopharyngeal squamous cell carcinoma, we conducted experiments on the functional study of miRNA-203 and identification of miRNA-203 regulated target genes in hypopharyngeal cancer cells. We found that cell proliferation and cell colony-forming increased more in the miRNA-203 up-regulated cancer cells than in the negative control cancer cells. Up-regulation of miRNA-203 accelerated cell cycle progression in hypopharyngeal cancer cells. TP63 and B3GNT5 mRNAs were identified and validated as targets of miRNA-203. However, transwell assay and wound scratch assay showed that miRNA-203 did not involve in invasion and metastasis in hypopharyngeal cancer cells. According to the results, we conclude that miRNA-203 can promote tumor growth in human hypopharyngeal squamous cell carcinoma. These results provide the convincing evidence for the first time that up-regulation of miRNA-203 contributes to the malignancy of hypopharyngeal squamous cell carcinoma, possibly through down-regulating TP63 and B3GNT5.

Background: Sinonasal squamous cell carcinoma (SSCC) and nasal inverted papilloma (NIP) represent the predominant type of malignant and benign tumors in sinonasal tract, respectively. CD4+ CD25+ Foxp3+ natural regulatory T (Treg) cells might play critical role(s) in the suppression of anti-tumor immune response and thus shed light on tumor progression from benign to malignant., Objective: This study aimed to evaluate the frequency and suppressive capacity of Treg cells in SSCC compared to NIP and further to explore the underlying mechanisms., Patients and Methods: Frequencies of Treg, Th1 and Th2 cells were evaluated by flow cytometry in tissue homogenate and peripheral blood from 31 SSCC patients, 32 NIP patients and 35 normal controls. Treg cells were tested for regulatory function by co-culture with effector T cells. CCR4 and its ligands, CCL22 and CCL17, were analyzed by flow cytometry and Luminex, respectively. The chemoattractant properties of CCR4/CCL22 and CCR4/CCL17 for Treg cells were assessed using the Boyden chamber technique, to elucidate the potential mechanisms of Treg recruitment in tumor microenvironment. Treg cells induction via TGF-β was assessed with transwells after local CD4+ Foxp3+ T cells were assessed by immunohistochemistry and TGF-β concentration was measured by Luminex., Results: Tumor-infiltrating Treg cells increased significantly from normal to NIP to SSCC (P ≤ 0.001 for normal vs. NIP and P = 0.004 for NIP vs. SSCC). Significantly elevated frequency and enhanced suppression capacity of circulating Treg cells in SSCC were detected compared to NIP and healthy controls, concomitant with Th1 decrease and Th2 increase. Apparently increased CCL22 attracted CCR4-expressing Treg cells to tumor microenvironment in SSCC, compared to NIP. SSCC produced significantly more TGF-β than NIP and thus possessed greater potential for Treg cell induction., Conclusion: Frequency and suppressive capacity of Treg cells enhanced with progression of malignancy from NIP to SSCC. Circulating Treg cells were recruited to tumor tissue via CCR4/CCL22 signalling, whereas tumor-synthesised TGF-β contributed to induction of peripheral Treg cells.

To identify genetic markers for laryngeal squamous cell carcinoma (LSCC), we conducted a genome-wide association study (GWAS) on 993 individuals with LSCC (cases) and 1,995 cancer-free controls from Chinese populations. The most promising variants (association P < 1 × 10(-5)) were then replicated in 3 independent sets including 2,398 cases and 2,804 controls, among which we identified 3 new susceptibility loci at 11q12 (rs174549), 6p21 (rs2857595) and 12q24 (rs10492336). The minor alleles of each of these loci showed protective effects, with odds ratios (95% confidence intervals) of 0.73 (0.68-0.78; P = 1.00 × 10(-20)), 0.78 (0.72-0.84; P = 2.43 × 10(-15)) and 0.71 (0.65-0.77; P = 4.48 × 10(-14)), respectively. None of these variants showed an interaction with smoking or drinking. This is the first GWAS to our knowledge solely on LSCC, and the findings might advance understanding of the etiology of LSCC.

Objective: To identify the genes differentially expressed and underlying molecular mechanism in laryngeal squamous cell carcinoma by using cDNA microarrays., Methods: Using Illumina Human HT-12 BeadChip, gene expressions were detected in ten pairs of laryngeal cancer tissues and adjacent normal tissues. Total RNA was extracted and reverse-transcribed into cDNA. Labeled cDNA were hybridized with cDNA microarray, data were read and images were scanned. All the samples had passed quality control testing., Results: Through Illumina Genomstudio 1.9.0 Data processing software (P < 0.05 or diffscore>13, diffscore<-13) and multiple displacement t test(FDR<0.05), 426 genes showed statistically significant differences in expressions between laryngeal tumor tissues and corresponding adjacent normal tissues, with 222 up-regulated genes and 204 down-regulated genes in laryngeal cancer tissues. These up- or down-regulated genes were indicated to involve in cellular processes relevant to the cancer phenotype, such as proliferation, cell cycle, chromosome segregation, mitosis and meiosis. These differentially expressed genes also took part in cancer related signaling pathways as well, for instance, metabolic pathways, cell cycle, DNA replication, glutathione metabolism, mucin type O-Glycan biosynthesis, drug metabolism-cytochrome P450 and so on., Conclusion: The set of genes identified here and their functional annotations contribute to a better understanding of the pathogenesis of laryngeal squamous cell carcinoma from the view of multiple gene interactions and provide candidate markers for improving diagnosis, prognosis and treatment of this cancer.

Background: Laryngeal squamous cell carcinoma (LSCC) is the most common type in head and neck squamous cell carcinoma (HNSCC), and the development and progression of LSCC are multistep processes accompanied by changes of molecular biology., Objective: The purpose of this study was to investigate the molecular basis of tumorigenesis and regional lymph node metastasis in LSCC, and provide a set of genes that may be useful for the development of novel diagnostic markers and/or more effective therapeutic strategies., Methods: A total number of 10 patients who underwent surgery for primary laryngeal squamous cell carcinoma were recruited for microarray analysis. LSCC tissues compared with corresponding adjacent non-neoplastic tissues were analysed by Illumina mRNA microarrays, and LSCC tissues with regional lymph node metastasis and LSCC tissues without regional lymph node metastasis were analyzed in the same manner. The most frequently differently expressed genes screened by microarrays were also validated by qRT-PCR in another 42 patients diagnosed for LSCC., Results: Analysed by Illumina mRNA microarrays, there were 361 genes significantly related to tumorigenesis while 246 genes significantly related to regional lymph node metastasis in LSCC. We found that the six genes (CDK1, CDK2, CDK4, MCM2, MCM3, MCM4) were most frequently differently expressed functional genes related to tumorigenesis while eIF3a and RPN2 were most frequently differently expressed functional genes related to regional lymph node metastasis in LSCC. The expressions of these genes were also validated by qRT-PCR., Conclusions: The research revealed a gene expression signature of tumorigenesis and regional lymph node metastasis in laryngeal squamous cell carcinoma. Of the total, the deregulation of several genes (CDK1, CDK2, CDK4, MCM2, MCM3, MCM4, EIF3a and RPN2) were potentially associated with disease development and progression. The result will contribute to the understanding of the molecular basis of LSCC and help to improve diagnosis and treatment.

ZFX (zinc finger protein, X-linked) gene locus on the human X chromosome is structurally similar to the zinc finger protein, Y-linked gene, which may constitute the primary sex-determining signal. However, the pathological roles of the dysfunction of ZFX gene in human disease such as cancer have not been addressed. Here, we analyzed the expression of ZFX in human laryngeal squamous cell carcinoma (LSCC) tissue specimens and found a significant up-regulation compared to corresponding non-tumorous LSCC tissue. Recombinant lentivirus expressing ZFX short hairpin RNA (shZFX) was constructed and infected Hep-2 human LSCC cells. We found that knockdown of ZFX gene resulted in suppression of proliferation and colony-forming ability of Hep-2 cells, and led to S phase cell cycle arrest. In addition, down-regulation of ZFX induced a significant enhancement of cell apoptosis and expression changes of apoptosis-related genes. These results suggest that high expression of ZFX is associated with LSCC progression and knockdown of ZFX may block tumor cell growth mainly by promoting cell apoptosis.

The purpose of this study was to examine whether functional polymorphisms in the cytochrome P450 (CYP) enzyme genes affect the risk of developing larynx and hypopharynx squamous cell carcinoma (SCC). We investigated CYP1A1, CYP1B1, CYP2E1, and CYP3A4 polymorphisms in 278 patients with laryngeal and hypopharyngeal SCC and 278 control subjects by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Subjects with the CYP1A1 3798CC or TC genotype had an odds ratio (OR) of 3.26 (95% confidence interval CI=1.76-6.03) or 1.56 (95% CI=1.06-2.31), compared with those with the TT genotype. An increased risk was also associated with the CYP1A1 462Val/Val genotype (OR=2.39, 95% CI=1.11-5.16), compared with the TT genotype. Haplotype analysis suggested a synergistic effect of these two polymorphisms. A multiplicative joint effect between the CYP1A1 3798T>C polymorphism and smoking was observed. The OR (95% CI) of the TC or CC genotype for nonsmokers and smokers of >20 pack-years were 1.85 (0.99-3.44) or 8.15 (4.35-15.26), respectively (P(trend)<0.05). The CYP1A1 single-nucleotide polymorphisms are associated with an increased risk of developing smoking-related laryngeal and hypopharyngeal SCC in a Han Chinese population.