Previous studies have found that miR-375 and miR-205 were significantly dysregulated in laryngeal squamous cell carcinoma, which contributed to the invasion and migration of LSCC. However, the mechanisms of miR-375 and miR-205 regulating the invasion and migration of LSCC remain unknown. qRT-PCR was performed in 40 pairs of tissue samples to investigate the expression of miR-375 and miR-205 in LSCC and paracarcinoma tissues. To investigate whether or not miR-375 and miR-205 regulated the invasion and migration of LSCC synergistically via AKT-mediated epithelial-mesenchymal transition, miR-375 mimic and miR-205 inhibitor were transfected into SNU899 cells and miR-375 inhibitor and miR-205 mimic were transfected into SNU899 cells, respectively, with or without AKT inhibitor. Then the expressions of miR-375 and miR-205 were validated by qRT-PCR, cell migration and invasion were determined by wound healing assay and transwell invasive assay, and western blot analysis was performed to detect the expression of related proteins. Our results showed that miR-375 and miR-205 regulated the invasion and migration of LSCC via AKT-mediated EMT synergistically. In conclusion, our findings provided not only new information about the molecular mechanism of miRNAs regulating invasion and migration of LSCC, but also a theoretical principle for potential targeting therapy of laryngeal squamous carcinoma.

BACKGROUND: The optimal management of glottic carcinoma involving the anterior commissure is controversial.
METHOD: A retrospective analysis was conducted of 76 patients with glottic squamous cell carcinoma treated by transoral carbon dioxide laser resection by a single surgeon.
RESULTS: Sixty-three patients (with tumour stage Tis-T3) were eligible for inclusion. Thirty patients had involvement of the anterior commissure; these patients were significantly more likely to have either uncertain or positive margins (63.3 vs 30.3 per cent, p = 0.012), and were also more likely to receive adjuvant radiotherapy (40 vs 3.2 per cent, p = 0.0005). The overall laryngeal preservation rate was 96.8 per cent; there was no statistically significant difference between those with and without anterior commissure involvement (96.7 and 96.9 per cent respectively).
CONCLUSION: Transoral laser resection with the use of adjuvant radiotherapy in a minority of patients with adverse pathological findings can be recommended for the primary treatment of anterior commissure glottic cancer from an oncological perspective; excellent local control and laryngeal preservation rates can be achieved.

OBJECTIVE: The aim of the present study was to characterize the pharyngoesophageal segment in laryngectomees who rated themselves as functional tracheoesophageal speakers.
METHODS: Voice perceptual assessment, high-resolution videomanometry of swallowing and phonation, and high-speed camera recording during phonation provided information about the anatomy and function of the pharyngoesophageal segment.
RESULTS: Fourteen patients were included in the study. The voice assessments presented high intra/inter-listener reliability. We found a significant correlation between roughness and poor voice quality, hyperfunction and poor intelligibility, and poor voice quality, long time since the operation, and old age. High-resolution videomanometry during phonation revealed decreasing mean pressures from the distal esophagus to the pharynx and confirmed low resting pressures at the pharyngoesophageal segment and low esophageal peristaltic contraction pressures after laryngectomy in comparison to normal subjects. The neoglottis shape was mainly circular and presented a strong mucosal wave in most of the patients on the high-speed camera recording.
CONCLUSIONS: Perceptual voice assessment and high-speed camera recordings provided baseline information about voice characteristics and vibration regularity of the neoglottis. Additionally, the quantitative measures obtained with high-resolution videomanometry may have clinical applicability as reference data in voice rehabilitation after total laryngectomy.

OBJECTIVE: This study assessed the relationship between vascular endothelial growth factor expression and the laryngeal cancer prognosis.
METHODS: Systematic computerised searches of PubMed were performed up to 31 January 2015. Prognostic endpoints were overall survival and disease-free survival. The pooled hazard ratios for overall survival and disease-free survival were also calculated.
RESULTS: Seven studies containing 975 patients were included. The pooled hazard ratio was 1.703 (95 per cent confidence interval, 1.373 to 2.112; z score = 4.85, p = 0.000) for overall survival and was 1.918 (95 per cent confidence interval, 1.410 to 2.609; z score = 4.15, p = 0.000) for disease-free survival. No significant publication bias was found. A sensitivity analysis showed that the results were robust. Power analyses also showed there was enough power to detect the calculated hazard ratios.
CONCLUSION: The study found that vascular endothelial growth factor overexpression predicted a worse prognosis for laryngeal cancer patients. This supports a strategy of targeted therapy by blocking the vascular endothelial growth factor receptor.

BACKGROUND: We reviewed the outcomes of patients with T3 laryngeal neoplasms with a fixed hemilarynx, a large gross tumor volume or a subglottic extension (SGE), treated with a laryngeal-preservation protocol with induction chemotherapy.
PATIENTS AND METHODS: The study end-points were laryngo-esophageal dysfunction-free survival (LEDFS), laryngectomy-free survival (LFS), overall survival (OS), and disease-free survival (DFS).
RESULTS: A total of 104 patients were included. The 2-year and 5-year OS rates were 70.4% and 54.5%, respectively. OS and DFS were independent of the treatment modality in the whole cohort (p=0.6546 and p=0.3006, respectively) and in patients with SGE (p=0.529 and p=0.255, respectively). The 2-year and 5-year LEDFS rates were 44.3% and 28.2%, respectively. LEDFS was not associated with initial hemilaryngeal fixation or SGE (p=0.5772 and p=0.0623, respectively).
CONCLUSION: Chemoselection is feasible without compromised oncological or functional outcomes in patients with an initially fixed hemilarynx or subglottic extension.

Chemokine and its receptors play important roles in laryngeal cancer development and progression. CCR6 is the receptor of CCL20 chemokine, but its function in laryngeal cancer is not known. The aim of this study is to explore the roles of CCR6 and its regulation mechanism in laryngeal cancer. We found CCR6 expression was higher in laryngeal cancer tissues compared with their normal controls, so did in laryngeal cancer cells. Cellular function indicated that down-regulation of CCR6 in laryngeal cancer cells could suppress cell proliferation and metastasis. Further research showed that CCR6 could activate p38, which was related with the changes of microRNA (miRNA) profile in laryngeal cancer cells. We also found that CCR6 was positively associated with miR-20a-5p, miR-489 and negatively related to miR-29-3p, miR-632 and miR-1276 in laryngeal cancer tissues.

Laryngeal squamous cell carcinoma (LSCC) is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. Runt-related transcription factor 3 (RUNX3), an important tumor suppressor, is known to related to lymph node metastasis and the development of LSCC. However, the biological roles and potential mechanisms RUNX3 expression was not well understood. In this study, we reported that the RUNX3 was significantly downregulated and highly methylated in LSCC compared with their matched normal. The enforced expression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. We identified that RUNX3 was regulated by miR-148a-3p and found that the expression level of miR-148-3p was significantly decreased and positively related with the expression of RUNX3 in LSCC. We also identified that DNA methyltransferase enzyme DNA (cytosine-5-)-methyltransferase 1 (DNMT1) was targeted by miR-148a-3p in LSCC. The knockdown of DNMT1 promoted the expression of RUNX3 and inhibited migration, invasion, and proliferation in LSCC cells. In summary, our study demonstrated that miR-148a-3p may regulate RUNX3 expression through the modulation of DNMT1-dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC. LSCC is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. In this study, we reported that RUNX3, an important tumor suppressor, was significantly downregulated and highly methylated in LSCC compared with their matched normal. The overexpression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. Moreover, RUNX3 was regulated by miR-148a-3p, which targeted DNA methyltransferase enzyme DNMT1 in LSCC cells. Therefore, miR-148a-3p may regulate RUNX3 expression through the modulation of DNMT1-dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC.

BACKGROUND/AIMS: This study aims to investigate the effect of CD47 on the development of laryngeal squamous cell carcinoma (LSCC) and the therapeutic potential of monoclonal antibody against CD47 and its ligand SIRPα in the treatment of LSCC.
METHODS: We firstly detected the expressions of CD47 mRNA and protein in LSCC and para-carcinoma tissues, introduced the most efficient CD47siRNA sequence into LSCC cells by lentiviral transfection and employed three monoclonal antibodies to evaluate their anti-LSCC effects in vitro and in vivo.
RESULTS: We observed that the mRNA and protein expressions of CD47 in LSCC tissue had significant increase in LSCC tissues compared with those in para-carcinoma tissue (p < 0.05). After the treatments of three monoclonal antibodies, i.e. anti-SIRPα, anti-CD47 BRIC126, anti-CD47 B6H12.2, in rats transfected with Hep-2 cell, it has been showed that the mRNA and protein expressions of CD47 in LSCC tissue decreased, macrophage efficiency was promoted when anti-SIRPα and/or CD47siRNA were used, the amounts, viabilities and expressions of CD47 protein of tumor cell were significantly inhibited. Additionally, combined use of CD47siRNA and anti-SIRPα seemed more efficient than solo use of CD47siRNA/anti-SIRPα.
CONCLUSION: The results suggested a critical role of CD47 in LSCC development and the promising treatment of antiCD47/SIRPα and/or CD47siRNA in LSCC.

This is the official guideline endorsed by the specialty associations involved in the care of head and neck cancer patients in the UK. Significantly new data have been published on laryngeal cancer management since the last edition of the guidelines. This paper discusses the evidence base pertaining to the management of laryngeal cancer and provides updated recommendations on management for this group of patients receiving cancer care. Recommendations • Radiotherapy (RT) and transoral laser microsurgery (TLM) are accepted treatment options for T1a-T2a glottic carcinoma. (R) • Open partial surgery may have a role in the management of selected tumours. (R) • Radiotherapy, TLM and transoral robotic surgery are reasonable treatment options for T1-T2 supraglottic carcinoma. (R) • Supraglottic laryngectomy may have a role in the management of selected tumours. (R) • Most patients with T2b-T3 glottic cancers are suitable for non-surgical larynx preservation therapies. (R) • Concurrent chemoradiotherapy should be regarded as the standard of care for non-surgical management. (R) • Subject to the availability of appropriate surgical expertise and multi-disciplinary rehabilitation services, TLM or open partial surgical procedures ± post-operative RT, may be also be appropriate in selected cases. (R) • In the absence of clinical or radiological evidence of nodal disease, elective treatment (RT or surgery ± post-operative RT) is recommended to at least lymph node levels II, III and IV bilaterally. In node positive disease, it is recommended that lymph node levels II-V should be treated on the involved side. If level II nodes are involved, then elective irradiation of ipsilateral level Ib nodes may be considered. (R) • Most patients with T3 supraglottic cancers are suitable for non-surgical larynx preservation therapies. (R) • Concurrent chemoradiotherapy should be regarded as the standard of care for non-surgical management. (R) • Subject to the availability of appropriate surgical expertise and multi-disciplinary rehabilitation services, TLM or open partial surgical procedures ± post-operative RT, may also be appropriate in selected cases. (R) • In the absence of clinical or radiological evidence of nodal disease, elective treatment (RT or surgery ± post-operative RT) is recommended to at least lymph node levels II, III and IV bilaterally. In node positive disease, lymph node levels II-V should be treated on the involved side. (R) • As per the PET-Neck clinical trial, patients with N2 or N3 neck disease who undergo treatment with chemoradiotherapy to their laryngeal primary and experience a complete response with a subsequent negative post-treatment positron emission tomography combined with computed tomography (PET-CT) scan do not require an elective neck dissection. In contrast, patients who have a partial response to treatment or have increased uptake on a post-treatment PET-CT scan should have a neck dissection. (R) • Larynx preservation with concurrent chemoradiotherapy should be considered for T4 tumours, unless there is tumour invasion through cartilage into the soft tissues of the neck, in which case total laryngectomy yields better outcomes. (R) • In the absence of clinical or radiological evidence of nodal disease, elective treatment (RT or surgery ± post-operative RT) is recommended to bilateral lymph node levels II, III, IV, V and VI. (R).

BACKGROUND/AIM: Surgery and chemotherapy treatments of human laryngeal squamous cell carcinoma (HLSCC) may fail due to metastasis, in which epithelial-mesenchymal transition (EMT) plays an important role. TRPP2, a nonselective cation channel, is expressed in various cell types and participates in many biological processes. Here, we show that TRPP2 enhanced metastasis by regulating EMT.
METHODS: We used immunohistochemistry, western blotting, Ca2+ imaging, transwell and wound healing assays to investigate TRPP2 expression levels in HLSCC tissue, and the role of TRPP2 in invasion and metastasis of a human laryngocarcinoma cell line (Hep2 cell).
RESULTS: We found that TRPP2 protein expression levels were significantly increased in HLSCC tissue; higher TRPP2 levels were associated with decreased patient survival time and degree of differentiation and advanced clinical stage. Knockdown of TRPP2 by transfection with TRPP2 siRNA markedly suppressed ATP-induced Ca2+ release, wound healing, and cell invasion in Hep2 cells. Moreover, TRPP2 siRNA significantly decreased vimentin expression but increased E-cadherin expression in Hep2 cells. In the EMT signalling pathway, TRPP2 siRNA significantly decreased Smad4, STAT3, SNAIL, SLUG and TWIST expression in Hep2 cells.
CONCLUSION: We revealed a previously unknown function of TRPP2 in cancer development and a TRPP2-dependent mechanism underlying laryngocarcinoma cell invasion and metastasis. Our results suggest that TRPP2 may be used as a biomarker for evaluating patient prognosis and as a novel therapeutic target in HLSCC.

OBJECTIVE: Complete separation of upper and lower respiratory tract after total laryngectomy results in permanent effects on nasal cavities and tracheo-bronchial airways. Aim of this study is evaluating nasal and tracheal cytological alterations of mucosa in laryngectomy long-term survivors, analyzing the feasibility of scraping for cytological examination of tracheal mucosa.
METHODS: Twenty-five laryngectomy patients underwent symptoms' evaluation, endoscopic fiber optic examination, prick tests, and nasal and tracheal scraping for cytological exam. Twenty-five healthy subjects underwent the same assessment, except for tracheal scraping. Eleven laryngectomy patients accepted inferior turbinate biopsy for histological examination.
RESULTS: Nasal cytological analysis demonstrated mucous cell metaplasia in 20% of laryngectomized patients, but it was absent in all healthy subjects; no squamous cell metaplasia was found in both groups. In 15 patients (60%), bacteria were present, without inflammatory infiltrate. Tracheal cytological analysis demonstrated a quite high rate of squamous cell metaplasia (24%), neutrophilic infiltrate (32%), and presence of bacteria (40%). Histological examination of inferior turbinate showed submucosal stromal fibrosis in all patients and submucosal inflammatory infiltrate in 1 case (9%).
CONCLUSION: Nasal cavities and trachea of laryngectomy patients undergo long-term cytological and histological changes of mucosa and submucosa, probably due to airflow modifications.

The purpose of this study was to evaluate the treatment of clinically negative cervical lymph nodes in supraglottic carcinoma by a meta-analysis. The search words were "supraglottic carcinoma", "cervical lymph nodes negative/cN0", "radical neck dissection", and "radiotherapy". The databases included the Chinese biomedical literature database, Medline, Cochrane library, EMBASE database, journals, and theses, etc. from 1989 onwards. Using the 5-year overall survival, disease-free survival, and disease-specific survival rates, and the recurrence and distant metastasis rates as observation indexes, the proper model and method were selected after a heterogeneity test to allow combined statistic tests, sensitivity analysis, and publication bias analysis to be conducted. Four studies (807 cases) were included in the analysis. Comparisons of the 5-year overall survival, disease-free survival, and disease-specific survival rates as well as lymph node metastasis and the recurrence rate for radical neck dissection and radiotherapy showed no significant differences. There was no advantage of radical neck dissection in supraglottic carcinoma with clinically negative cervical lymph nodes compared to radiotherapy. However, owing to the lack of a prospective study and large number of cases, selection bias and measurement bias may still exist.

Laryngeal cancer is the major malignant tumor affecting the upper respiratory tract. Previous studies have reported on the association between XRCC1 genetic polymorphisms and risk of laryngeal cancer, but with conflicting results. In this study, we attempted to assess the association between XRCC1 Arg194Trp, Arg280His and Arg399Gln polymorphisms and risk of laryngeal cancer in a Chinese population. A total of 126 laryngeal cancer patients and 254 control subjects were recruited to this study from the Second Medical College of Jinan University between December 2013 and May 2015. The XRCC1 Arg194Trp, Arg280His, and Arg399Gln polymorphic sites were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Our results revealed a significant association between the AA genotype of XRCC1 Arg280His [odds ratio (OR) = 2.51, 95% confidence interval (CI) = 1.29-4.87, P = 0.01] and an increased risk of laryngeal cancer susceptibility compared to the GG genotype. Moreover, the A allele showed a higher risk of laryngeal cancer susceptibility compared to the G allele (OR = 1.63, 95%CI = 1.19-2.50, P = 0.002). In conclusion, the results of our study suggest that the AA genotype and A allele of the XRCC1 Arg280His polymorphism are associated with an increased laryngeal cancer risk in a Chinese population.

BACKGROUND/AIM: Recurrent laryngeal cancer often shows an aggressive phenotype after radiotherapy and does not respond to conventional therapeutic strategies. In this study, we investigated the contribution of furin to cellular invasiveness in radio-resistant laryngeal cancer.
MATERIALS AND METHODS: Using previously established AMC-HN-3 and AMC-HN-8 cell lines from laryngeal carcinoma patients, recurrent laryngeal cancer models were generated by cumulative irradiation (AMC-HN-3-70Gy and AMC-HN-8-70Gy). Immunocytochemistry and western blotting were used to determine the epithelial-mesenchymal transition (EMT). Invasion capacity was assessed using an in vitro invasion assay. Zymography was used to assess metalloproteinase-2 (MMP-2) activity. Tumor xenografts were developed to compare growth rate and furin expression in vivo. Furin expression in 35 patients (45 samples) with salvage total laryngectomy after radiation-based treatment was assessed by laryngeal cancer tissue microarray.
RESULTS: Both AMC-HN-3-70Gy and AMC-HN-8-70Gy cell lines underwent EMT following radiation. However, AMC-HN-3-70Gy cells showed increased cellular invasiveness, whereas AMC-HN-8-70Gy cells showed no difference. AMC-HN-3-70Gy cells also exhibited elevated furin expression with up-regulated expression of the active form of membrane type 1-matrix metalloproteinase (MT1-MMP)/MMP-2, whereas AMC-HN-8-70Gy cells did not show significant changes. After administration of a furin inhibitor (chloromethyl ketone (CMK)), AMC-HN-3-70Gy cells showed a significant decrease in MT1-MMP/MMP-2 expression and cellular invasiveness. Nine of 22 samples (40.9%) from salvage total laryngectomy and one of 13 pre-radiation samples (7.7%) had high furin expression. Post-radiation, furin expression increased in seven of 10 patients whose pre- and post-radiation samples were available; all-cancer mortality (three patients) was observed in this group.
CONCLUSION: Together with EMT, furin activity may serve as an indicator of an aggressive cancer phenotype, suggesting that furin is a potentially useful target for recurrent laryngeal cancer.

BACKGROUND: In this study, we examined epidemiological aspects of dynamic changes in incidences of laryngeal cancer in male and female populations in Kazakhstan.
MATERIALS AND METHODS: Primary data were for registered patients with malignant laryngeal tumors in the whole country during the period of 1999-2014. Evaluation of changes in laryngeal cancer incidence in the population of Kazakhstan was performed using component analysis.
RESULTS: It was determined that the number of patients with laryngeal cancer in the whole country is decreasing although with conflicting impacts of different factors. Despite population growth (all - ΔP=+66.1%, men - ΔP=+70.9% and women - ΔP=+46.4%), and aging (all - ΔA=+45.1%, men -ΔA=+54.3 and women - ΔA=+22.2), the reduction in risk of developing the disease (all - ΔR=-165.6%, men - ΔR=-170.9% and women - ΔR=-141.0%) was overwhelming.
CONCLUSIONS: This investigation was the first epidemiological study of dynamics of laryngeal cancer by component analysis in the population of Kazakhstan. Implementation of the results of the study is recommended in management of anti-cancer activities for laryngeal cancer.

BACKGROUND AND OBJECTIVES: Previous studies showed good short-term Quality of life (QOL) after Transoral Laser Microsurgery (TLM) for laryngeal cancer. Here, we aimed to evaluate QOL after TLM in the long-term.
METHODS: Prospective longitudinal study. Sixty-two consecutive disease-free patients were evaluated using UW-QOL v4 and SF-12 questionnaires, 1 and 5 years after TLM. Changes over time were assessed according to age, location, and tumor size. Long-term VHI-10 was also evaluated.
RESULTS: The mean follow-up time was 5.41 ± 2.02 years. No differences in the global UW-QOL score were observed between 1 and 5 years after TLM (1135.00 vs. 1127.20; P = 0.4). Activity worsened slightly in the long-term (93.03 vs. 87.70; P = 0.02). Forty-two and 58% of the patients reported that their health 1 and 5 years after treatment was much better than prior to diagnosis. Initially, 3.3% considered their health much worse, which was reduced to 1.7% at 5 years. SF-12 scores remained unchanged for both physical and mental aspects (P > 0.05). The VHI-10 was 3.81 ± 5.7 for supraglottic and 7.2 ± 9.6 for glottic tumors.
CONCLUSION: Patients treated with TLM present a very good long-term QOL. Only activity deteriorates over time, while voice and swallowing remain satisfactory in the majority of patients. J. Surg. Oncol. 2016;114:789-795. © 2016 2016 Wiley Periodicals, Inc.

BACKGROUND: Nodal metastasis is an important prognostic factor in head and neck squamous cell carcinoma. This study aimed to determine the average nodal basin yield per level of neck dissection, and to investigate if age, gender, body mass index, tumour size, depth of tumour invasion and p16 status influence nodal yield.
METHOD: A retrospective review of 185 patients with head and neck squamous cell carcinoma generated 240 neck dissection specimens.
RESULTS: The respective mean nodal yields for levels I, II, III, IV and V were 5.27, 9.43, 8.49, 7.43 and 9.02 in non-cutaneous squamous cell carcinoma patients, and 4.2, 7.57, 9.65, 4.33 and 12.29 in cutaneous squamous cell carcinoma patients. Multiple regression analysis revealed that p16-positive patients with mucosal squamous cell carcinoma yielded, on average, 2.4 more nodes than their p16-negative peers (p = 0.04, 95 per cent confidence interval = 0.116 to 4.693). This figure was 3.84 (p = 0.008, 95 per cent confidence interval = 1.070 to 6.605) for p16-positive patients with oral cavity squamous cell carcinoma.
CONCLUSION: In mucosal squamous cell carcinoma, p16-positive status significantly influenced nodal yield, with the impact being more pronounced in oral cavity squamous cell carcinoma patients.

OBJECTIVES: 8-Hydroxy-2'-deoxyguanosine is a biomolecule associated with DNA damage. We evaluated oxidative stress and DNA damage in patients with laryngeal cancer by measuring 8-hydroxy-2'-deoxyguanosine levels.
METHODS: This study enrolled 117 subjects, including 64 controls and 53 patients who had benign vocal cord lesions or laryngeal cancer. The benign excised lesions, tumor tissue, noncancerous laryngeal tissue, blood, and urine were subjected to high-performance liquid chromatography, and 8-hydroxy-2'-deoxyguanosine levels were compared between groups.
RESULTS: Blood and urine 8-hydroxy-2'-deoxyguanosine levels in patients with laryngeal carcinoma were significantly higher than in the controls ( P = .00002, P = .00001). The 8-hydroxy-2'-deoxyguanosine level was significantly higher in tumor tissues than in non-tumor tissue and benign vocal cord lesion tissues ( P = .00002, P = .000001).
CONCLUSIONS: We determined that laryngeal cancer was associated with oxidative stress, which may be quantified by measuring 8-hydroxy-2'-deoxyguanosine. For a patient with a suspicious laryngeal lesion, 8-hydroxy-2'-deoxyguanosine levels in blood and urine can provide advance information about the likely diagnosis.

OBJECTIVE: To evaluate the clinical and histopathological factors affecting the prognosis of patients with squamous cell locoregional advanced laryngeal cancer.
METHODS: A retrospective chart review was conducted of 121 patients with locoregional advanced laryngeal cancer, primarily treated with surgery from 2007 to 2011. Disease-free survival and overall survival rates were analysed as oncological outcomes. Prognostic variables, namely gender, pharyngeal invasion, pathological assessment of tumour and nodal stage, adjuvant therapy, margin status, nodal extracapsular extension, tumour differentiation, lymphovascular and perineural invasion, and predominant growth pattern, were also analysed.
RESULTS: One-year and three-year disease-free survival rates were 81.3 per cent and 63.5 per cent, respectively. One-year and three-year overall survival rates were 88.3 per cent and 61.4 per cent, respectively. Multivariate analysis showed that nodal extracapsular extension (p < 0.05) and an infiltrative growth pattern (p < 0.05) were associated with disease progression. Nodal extracapsular extension (p < 0.05) was associated with higher mortality.
CONCLUSION: Nodal extracapsular extension and an infiltrative growth pattern were the main prognostic factors in locoregional advanced laryngeal cancer. The presence of pharyngeal invasion, pathologically confirmed node-positive stage 2-3 disease, close or microscopic positive margins, and lymphovascular and perineural invasion have a negative impact on prognosis.

Targeting cancer cells is crucial for improving the efficiency of laryngeal cancer treatment. However, the signaling pathway and therapeutic strategy, related to the tumor, still need further research. Dietary flavonoid fisetin (3,3',4',7-tetrahydroxyflavone) found in many fruits and vegetables has been shown in preclinical studies to inhibit cancer growth through regulating cell cycle, apoptosis, angiogenesis, invasion and metastasis without causing any toxicity to normal cells. PI3K/AKT and ERK1/2 have been known as essential signaling pathways to modulate cell proliferation, apoptosis as well as autophagy via mTOR, Caspase-3 and NF-κB signals. In our study, flow cytometry and western blot assays suggested that apoptosis was induced by fisetin administration, promoting Caspase-3 expressions by regulating PI3K/AKT/NF-κB. Additionally, fisetin suppressed TU212 cells proliferation, which was linked with ERK1/2 inactivation. Further, the activation of PI3K/AKT-regulated mTOR was inhibited by fisetin, leading to transcription suppression and proliferation inhibition of TU212 cells. In vivo studies also showed that the tumor volume and weight of nude mice were reduced for fisetin use with KI-67 decrease and LC3II increase in tumor tissue samples. Together, our data indicated that fisetin had a potential role in controlling human laryngeal cancer through inhibiting tumor cell proliferation, inducing apoptosis and autophagy regulated by ERK1/2 and AKT/NF-κB/mTOR signaling pathways, which might provide a therapeutic strategy for laryngeal cancer inhibition in future.

OBJECTIVE: CKLF-like MARVEL transmembrane domain containing 3 (CMTM3), as a tumor suppressor gene, plays an important role in the suppression of cell growth and apoptosis. The goal of our study is to investigate the association between CMTM3 promoter methylation and laryngeal squamous cell carcinoma (LSCC).
DESIGN AND METHODS: Using the bisulfite pyrosequencing technology, DNA methylation levels of seven CpG sites in CMTM3 promoter are measured in tumor tissues and their adjacent tissues of 76 male LSCC patients.
RESULTS: Our results reveal a significantly elevated promoter methylation of CMTM3 in tumor tissues compared with their adjacent tissues (P<0.001). A breakdown analysis by age shows that significant association of CMTM3 promoter methylation with cancer risk is specific to the LSCC patients older than 55years (P<0.001) but not in the younger patients (P=0.305). Moreover, the association is only observed in the LSCC patients with smoking behavior (P=0.001). Breakdown analysis also shows that CMTM3 promoter methylation is associated with cancer risk among patients with stage I LSCC (P<0.001).
CONCLUSION: In conclusion, our study indicates that elevated CMTM3 methylation is a risk factor in male LSCC patients, especially in the patients with age over 55years and with smoking behavior.

OBJECTIVES: This study aimed to compare the diagnostic effectiveness of narrow band imaging and autofluorescence imaging for malignant laryngopharyngeal tumours.
METHODS: Between May 2010 and October 2010, 50 consecutive patients with suspected laryngopharyngeal tumour underwent endoscopic laryngopharynx examination. The morphological characteristics of laryngopharyngeal lesions were analysed using high performance endoscopic systems equipped with narrow band imaging and autofluorescence imaging modes. The diagnostic effectiveness of white light image, narrow band imaging and autofluorescence imaging endoscopy for benign and malignant laryngopharyngeal lesions was evaluated.
RESULTS: Under narrow band imaging endoscopy, the superficial microvessels of squamous cell carcinomas appeared as dark brown spots or twisted cords. Under autofluorescence imaging endoscopy, malignant lesions appeared as bright purple. The sensitivity of malignant lesion diagnosis was not significantly different between narrow band imaging and autofluorescence imaging modes, but was better than for white light image endoscopy (χ2 = 12.676, p = 0.002). The diagnostic specificity was significantly better in narrow band imaging mode than in both autofluorescence imaging and white light imaging mode (χ2 = 8.333, p = 0.016).
CONCLUSION: Narrow band imaging endoscopy is the best option for the diagnosis and differential diagnosis of laryngopharyngeal tumours.

MicroRNAs (miRNAs) play important roles in gene regulation during laryngocarcinoma. MiR-9 is a potential oncomiR, but its function in laryngocarcinoma is not known. The aim of this study is to investigate the roles of miR-9 in laryngocarcinoma. We found miR-9 expression was higher in laryngocarcinoma tissues compared with their normal controls, so did the laryngocarcinoma cells. Cellular function of miR-9 indicated that miR-9 restoration in laryngocarcinoma cells could promote cell proliferation and metastasis. Phosphatase and tensin homolog (PTEN) was predicted as a target gene of miR-9 and verified using luciferase reporter assay. PTEN expression was down-regulated in the laryngocarcinoma cells with miR-9 overexpression. We also found that miR-9 expression was negatively associated with PTEN expression in laryngocarcinoma tissues.

CONCLUSIONS: The results suggest that gender-specific differences in health-related quality-of-life (HRQoL) exist in patients with larynx carcinoma. In previous studies these differences might have been concealed by predominantly male subject groups. Future studies should consider a gender-specific analysis that suits the patient's idiosyncrasies associated with laryngeal cancer.
OBJECTIVES: There is little research concerning gender differences in quality-of-life (QoL) in patients with larynx carcinoma. Since laryngeal cancer is predominantly found in males, most studies examining HRQoL are based on a mainly male subject group. HRQoL needs to be assessed to determine the impact of disease and treatment and to evaluate possible treatment regimes. This study examined gender differences concerning HRQoL in 53 patients using EORTC QLQ-C30, and QLQ-H&N35 questionnaires.
METHODS: Patients treated with larynx carcinoma were given two questionnaires to assess HRQoL. The questionnaires were analyzed for each sex separately, as well as for the entire population.
RESULTS: Female patients report significantly worse HRQoL than males. Age could not be identified as a significant predictor for HRQoL when males and females were analyzed together, and does not significantly predict HRQoL in men. However, age was found to be a significant predictor for HRQoL when only females were analyzed.

Laryngeal squamous cell carcinoma (LSCC) is a very aggressive cancer, considered to be a subtype of the head and neck squamous cell carcinoma (HNSCC). Despite significant advances in the understanding and treatment of cancer, prognosis of patients with LSCC has not improved recently. In the present study, we sought to understand better the genetic mechanisms underlying LSCC development. Thirty-two tumor samples were collected from patients undergoing surgical resection of LSCC. The samples were submitted to whole-genome cDNA microarray analysis aiming to identify genetic targets in LSCC. We also employed bioinformatic approaches to expand our findings using the TCGA database and further performed functional assays, using human HNSCC cell lines, to evaluate viability, cell proliferation, and cell migration after silencing of selected genes. Eight members of the homeobox gene family (HOX) were identified to be overexpressed in LSCC samples when compared to normal larynx tissue. Quantitative RT-PCR analysis validated the overexpression of HOX gene family members in LSCC. Receiver operating characteristic (ROC) statistical method curve showed that the expression level of seven members of HOX gene family can distinguish tumor from nontumor tissue. Correlation analysis of clinical and gene expression data revealed that HOXC8 and HOXD11 genes were associated with the differentiation degree of tumors and regional lymph node metastases, respectively. Additionally, siRNA assays confirmed that HOXC8, HOXD10, and HOXD11 genes might be critical for cell colony proliferation and cell migration. According to our findings, several members of the HOX genes were overexpressed in LSCC samples and seem to be required in biological processes involved in tumor development. This suggests that HOX genes might play a critical role in the physiopathology of LSCC tumors.

BACKGROUND: Multi-step cancerogenesis guides laryngeal cancer onset and it includes a wide variety of pre-cancerous lesions macroscopically challenging to identify and distinguish from initial cancerous foci.
OBJECT: Different modalities of diagnostic techniques of laryngeal epithelial lesions exist and they do not offer a single system to make a differential diagnosis. Hence, this meta-analysis aimed to synthesize the validity of each single diagnostic tool to improve laryngeal patient management.
METHODS: A systematic review of literature was led searching for articles mentioning the following terms: larynx, laryngeal precancerous lesions, laryngeal cancer, white light (WL) endoscopy, stroboscopy, contact endoscopy (CE), autofluorescence (AF), ultrasound (US), narrow band imaging (NBI), computed axial tomography (CAT), magnetic resonance imaging (MRI), positron emission tomography (PET), CAT/PET. Then, a quantitative analysis was carried on for paper published after 2005 onward.
RESULTS: The search identified 7215 publications, of which 3616 published after 2005, with a final results of a total of 214 articles stratified and included by our selection criteria. 42 out of 214 articles were selected for quantitative synthesis. 25 out of 41 studies had a good quality score, 16 were fair.
CONCLUSIONS: A comprehensive overview of the most recent advances in laryngeal imaging technology combined with all of the information needed to interpret findings and manage patients with voice disorders can be found herein. Our flow-chart allows clinicians in risk-stratify patients and select proper examination modalities to provide appropriate care. Study limitations, together with possible clinical and research implications have been counted.

BACKGROUND: Validation of magnetic resonance imaging (MRI) and development of guidelines for the delineation of the gross tumor volume (GTV) is of utmost importance to benefit from the visibility of anatomical details on MR images and to achieve an accurate GTV delineation. In the ideal situation, the GTV delineation corresponds to the histopathologically determined 'true tumor volume'. Consequently, we developed guidelines for GTV delineation of laryngeal and hypopharyngeal tumors on MRI and determined the accuracy of the resulting delineation of the tumor outline on histopathology as gold standard.
MATERIAL AND METHODS: Twenty-seven patients with T3 or T4 laryngeal/hypopharyngeal cancer underwent a MRI scan before laryngectomy. Hematoxylin and eosin sections were obtained from surgical specimens and tumor was delineated by one pathologist. GTV was delineated on MR images by three independent observers in two sessions. The first session (del1) was performed according to clinical practice. In the second session (del2) guidelines were used. The reconstructed specimen was registered to the MR images for comparison of the delineated GTVs to the tumor on histopathology. Volumes and overlap parameters were analyzed. A target margin needed to assure tumor coverage was determined.
RESULTS: The median GTVs (del1: 19.4 cm(3), del2: 15.8 cm(3)) were larger than the tumor volume on pathology (10.5 cm(3)). Comparable target margins were needed for both delineation sessions to assure tumor coverage. By adding these margins to the GTVs, the target volumes for del1 (median: 81.3 cm(3)) were significantly larger than for del2 (median: 64.2 cm(3)) (p ≤ 0.0001) with similar tumor coverage.
CONCLUSIONS: In clinical radiotherapy practice, the delineated GTV on MRI is twice as large as the tumor volume. Validated delineation guidelines lead to a significant decrease in the overestimation of the tumor volume.

BACKGROUND: Selenium-binding protein 1 (SELENBP1) expression is reduced markedly in many types of cancers and low SELENBP1 expression levels are associated with poor patient prognosis.
METHODS: SELENBP1 gene expression in head and neck squamous cell carcinoma (HNSCC) was analyzed with GEO dataset and characteristics of SELENBP1 expression in paraffin embedded tissue were summarized. Expression of SELENBP1 in nasopharyngeal carcinoma (NPC), laryngeal cancer, oral cancer, tonsil cancer, hypopharyngeal cancer and normal tissues were detected using immunohistochemistry, at last, 99 NPC patients were followed up more than 5 years and were analyzed the prognostic significance of SELENBP1.
RESULTS: Analysis of GEO dataset concluded that SELENBP1 gene expression in HNSCC was lower than that in normal tissue (P < 0.01), but there was no significant difference of SELENBP1 gene expression in different T-stage and N-stage (P > 0.05). Analysis of pathological section concluded that SELENBP1 in the majority of HNSCC is low expression and in cancer nests is lower expression than surrounding normal tissue, even associated with the malignant degree of tumor. Further study indicated the low SELENBP1 expression group of patients with NPC accompanied by poor overall survival and has significantly different comparing with the high expression group.
CONCLUSION: SELENBP1 expression was down-regulated in HNSCC, but has no associated with T-stage and N-stage of tumor. Low expression of SELENBP1 in patients with NPC has poor over survival, so SELENBP1 could be a novel biomarker for predicting prognosis.

Here we describe the long-term outcomes of type I thyroplasty (TP-I) with silicone block implantation through histopathological assessments in a male patient who underwent pharyngolaryngectomy for secondary hypopharyngeal carcinoma 7 years after silicone implantation. A 66-year-old man presented with esophageal carcinoma and underwent subtotal esophagotomy. Subsequently, his left vocal fold exhibited fixation in a paramedian position, and he underwent TP-I with silicone block implantation 2 years after the primary esophageal surgery. His voice quality improved; however, he developed glottic carcinoma in the right vocal fold 6 months after TP-I and underwent laser cordectomy. Glottic carcinoma recurred 21 months later, and he underwent laser cordectomy again. Five years after the second laser surgery, he underwent pharyngolaryngectomy and neck dissection for hypopharyngeal carcinoma detected in the right pyriform sinus. We histopathologically examined a horizontal section of the resected larynx to assess silicone implant-related changes. Although migration of the silicone implant was not observed, a very mild foreign body reaction occurred around the implant. The patient is currently in remission. Our findings suggest that silicone implants are suitable for TP-I due to their remarkable affinity for human tissue and the low risk of a tissue reaction.

BACKGROUND: The small heat shock protein 27 kDA (Hsp27) acts as an ATP-independent chaperone in protein folding, but is also implicated in architecture of the cytoskeleton, cell migration, metabolism, cell survival, growth/differentiation, mRNA stabilization, and tumor progression.
OBJECTIVE: To study the intracellular localization of phosphorylated and non-phosphorylated forms of Hsp27 in squamous cell carcinoma of the larynx (SCCL) and to evaluate their relationship with regional lymphatic metastasis and overall five-year survival.
METHODS: Tumor biopsies of larynx tissue were collected from 50 patients who were between the ages of 30 to 80 years and had a confirmed diagnosis of squamous cell carcinoma of the larynx. Immunohistochemistry was used to determine the intracellular localization of the phosphorylated and non-phosphorylated forms of Hsp27.
RESULTS: The study revealed that the Hsp27 chaperone was expressed in both the cytoplasm and the nucleus of tumor cells in SCCL. The biopsies of patients with lymph node metastases showed significantly higher expression of the phosphorylated and unphosphorylated forms of Hsp27 in the nucleus compared to those of patients without lymph node metastases. At the same time, the cytoplasmic expression of Hsp27 in these patients did not differ statistically. Analysis of the overall five-year survival rates showed that negative Hsp27 expression in the nucleus of tumor cells is associated with the survival rate of patients with SCCL.
CONCLUSION: The nuclear expression of phosphorylated and unphosphorylated forms of Hsp27 is a molecular marker of unfavorable squamous cell carcinoma of the larynx associated with lymphogenous metastasis and decreased total five-year survival.