OTX1 exerts an oncogenic role and is negatively regulated by miR129-5p in laryngeal squamous cell carcinoma.

BACKGROUND: Orthodenticle homeobox 1 (OTX1) is a transcription factor that plays an important role in various human cancers. However, the function of OTX1 in laryngeal squamous cell carcinoma (LSCC) is largely unknown. We aimed to explore the roles of OTX1 in LSCC and its possible molecular mechanism. METHODS: The expression levels of OTX1 were assessed in LSCC cell lines and tissue samples. We further examined the effect of OTX1 on LSCC progression. The upstream regulator of OTX1 was identified using a computer algorithm and confirmed experimentally. RESULTS: OTX1 was highly expressed in 70.7% (70/99) of LSCC tissue samples. The OTX1 expression in LSCC was significantly correlated with lymph node metastasis. High OTX1 expression in patients with LSCC was correlated with poor prognosis. Knockdown of OTX1 inhibited proliferation, colony formation, migration and invasion in LSCC cells. Knockdown of OTX1 inhibited tumor growth in a xenograft mouse model. Mechanistically, OTX1 might act as a direct target of miR-129-5p. OTX1 enhanced tumorigenicity and tumor growth both in vitro and in vivo. CONCLUSIONS: Our findings support that OTX1 is an oncogene in LSCC tumorigenesis and progression. Furthermore, OTX1 is a direct target of miR-129-5p in LSCC cells. Taken together, OTX1 is a promising diagnostic and therapeutic marker for LSCC.

miR-144-3p, a tumor suppressive microRNA targeting ETS-1 in laryngeal squamous cell carcinoma.

Regional lymph node metastasis and distant metastasis are critical in the prognosis of laryngeal squamous cell carcinoma (LSCC). This study investigated the roles of miR-144-3p and E26 transformation specific-1 (ETS-1) in the invasion and migration of LSCC cells. The effects of miR-144-3p and ETS-1 on FaDu and Hep2 cell growth, migration and invasion were determined. Suppression of ETS-1 by miR-144-3p was confirmed using luciferase assays; the effects of ETS-1 silencing were determined using a xenograft tumor model. The expression of ETS-1 was analyzed in 71 paraffin-embedded tissue biopsies and eight fresh frozen biopsies obtained from LSCC patients. miR-144-3p inhibited the growth, invasion and migration of FaDu and Hep2 cells in part through suppression of epithelial-mesenchymal transition as determined by increased E-cadherin and α-catenin and reduced fibronectin and vimentin expression. Additionally, ETS-1 is a molecular target of miR-144-3p, and silencing ETS-1 expression inhibited FaDu and Hep2 cell invasion and migration as well as reduced Hep2 xenograft tumor volume. In LSCC, the expression of ETS-1 is upregulated with disease progression, and higher ETS-1 expression, which was negatively associated with miR-144-3p levels, adversely corresponded with prognoses. Thus, upregulated ETS-1 levels may promote LSCC metastasis, resulting in poor patient prognosis.

Preoperative neutrophil-to-lymphocyte ratio is an independent prognostic marker in patients with laryngeal squamous cell carcinoma.

BACKGROUND: Neutrophil-lymphocyte ratio (NLR) has been shown to be associated with prognosis in various solid tumors. This study aimed to evaluate the prognostic role of NLR in patients with laryngeal squamous cell carcinoma (LSCC). METHODS: A total of 141 LSCC patients were retrospectively reviewed. Patients' demographics were analyzed along with clinical and pathologic data. The optimal cutoff value of NLR was determined using receiver operating characteristic (ROC) curve analysis. The impact of the NLR and other potential prognostic factors on disease-free survival (DFS) and overall survival (OS) was assessed using the Kaplan-Meier method and multivariate Cox regression analysis. RESULTS: The optimal cutoff value of the NLR was 2.17. In the NLR ≤ 2.17 group, the 1-, 3-, and 5-year DFS rates were 88.2, 73.9 and 69.1 %, respectively, while in the NLR > 2.17 group, the DFS rates were 83.0, 54.6 and 49.2 %, respectively. Correspondingly, the 1-, 3-, and 5-year OS rates were 98.9, 85.1 and 77.4 % in the NLR ≤ 2.17 group and 97.9, 63.8 and 53.3 % in the NLR > 2.17 group, respectively. The multivariate Cox proportional hazard model analysis showed that NLR > 2.17 was a prognostic factor for both DFS [hazard ratio (HR) = 1.869; 95 % confidence interval (CI) 1.078-3.243; P = 0.026] and OS (HR =2.177; 95 % CI 1.208-3.924; P = 0.010). CONCLUSION: Our results showed that elevated preoperative NLR was an independent predictor of poor prognosis for patients with LSCC after surgical resection.

Effectiveness of chemotherapy and radiotherapy for laryngeal preservation in advanced laryngeal cancer: a meta-analysis and systematic review.

PURPOSE: This meta-analysis compared the efficacy of laryngectomy and three larynx-preservation treatments in patients with advanced laryngeal cancer. MATERIALS AND METHODS: Medline, Cochrane, EMBASE and Google Scholar databases were searched (until May 5, 2014) for studies that evaluated total laryngectomy followed by radiation and three larynx-preserving strategies in patients with advanced laryngeal cancer. The outcomes assessed were the 2- to 5-year overall survival (OS), disease-free survival (DFS), and laryngectomy-free survival (LFS) RESULTS: The search identified ten studies which were used for the meta-analysis (N = 2013 patients). The meta-analysis found that among different treatments, RT alone was associated with the highest rate of OS (71.6 %), DFS (57.8 %), and LFS (79.0 %). OS was similar among the following treatments: laryngectomy→RT vs. CT→RT [P = 0.402]; RT+CT vs. RT alone [P = 0.440]; and RT+CT vs CT→RT [P = 0.588]. DFS rate was higher in patients receiving laryngectomy→RT than CT→RT (P = 0.001) and was similar for RT+CT compared with CT→RT (P = 0.590). CONCLUSION: These findings suggest that of the larynx-preserving treatments, RT alone was associated with highest rates of OS, DFS, and LFS in patients with locally advanced cancer of the larynx and hypopharynx. Laryngectomy provided a better rate of DFS than CT and RT, but OS were similar across the different larynx-preserving treatments and laryngectomy.

Micro-ribonucleic acid expression profiling and bioinformatic target gene analyses in laryngeal carcinoma.

Abnormal expression of micro-ribonucleic acid (miRNA) might be clinically valuable as a biomarker or treatment target in the early diagnosis, treatment, and prognosis of tumors. However, little is known concerning abnormal miRNA expression of laryngeal carcinoma, one of the most commonly encountered head and neck tumors. Microarray analysis was used to obtain miRNA-expression profiles of ten pairs of freshly frozen laryngeal carcinoma tissue and surrounding normal tissue specimens. Characteristic miRNAs that were significantly related to laryngeal carcinoma were identified. Verification was performed using an additional 32 pairs of samples. The expression of two miRNAs (miR-21-3p and miR-106b-3p) was upregulated in both microarray and quantitative real-time polymerase chain-reaction analyses, whereas the expression of six miRNAs (let-7f-5p, miR-10a-5p, miR-125a-5p, miR-144-3p, miR-195-5p, and miR-203) was downregulated. The decreased expression of let-7f-5p and miR-195-5p is a novel finding in head and neck cancer. The target genes of these miRNAs were also predicted through multiple software programs. The differential expression of miRNAs might be related to the early onset and development of laryngeal carcinoma, and may be exploited as new biomarkers and therapeutic targets in the treatment of laryngeal carcinoma.

Retrospective analysis of prognostic factors in 205 patients with laryngeal squamous cell carcinoma who underwent surgical treatment.

OBJECTIVES: To investigate the most important factors affecting the prognosis of the patients with squamous cell carcinoma (SCC) of the larynx. METHODS: Based on the clinical and follow-up data, 205 patients with SCC of the larynx receiving total laryngectomy, partial laryngectomy, or CO2 laser surgery in GuangDong General Hospital were retrospectively analyzed. A survival analysis was performed by the Kaplan-Meier method and a multivariable analysis of prognostic factors was carried out using the Cox proportional hazard model. RESULTS: Subtypes of carcinoma included 69.8% glottic and 30.2% supraglottic. Most patients were in N0 stage (77.6%), and 22.4% patients were in N1∼N3 stage. Over half of the patients were in T1∼T2 stage (55.1%), 20.0% in T3, and 24.9% in T4. Mean follow-up duration was 49.2 months. The survival rates 1, 2, and 3 years after the surgery were 99.0%, 91.7%, and 81.5%, respectively. The survival rate for those patients with clinical stage IV was significantly lower than for those with clinical stage I and II (p<0.001 and p = 0.013, respectively). The disease-free progression rates 1, 2, and 3 years after the surgery were 83.9%, 74.6%, and 71.2%, respectively. Futhermore, those patients with a Charlson score of 1 to 2 and ≥3 had higher risk of mortality than those with a Charlson score of 0 (hazard ratios of 1.8 and 2.41 p = 0.042 and p = 0.008). Multivariable analysis revealed that clinical stage, surgical margin, and comorbidity were significantly associated with both mortality and disease-free progression. CONCLUSION: The surgical resection margin, clinical stage, and comorbidity were independent factors affecting the laryngeal cancer prognosis. The survival rates were lower for patients with advanced laryngeal cancer, positive surgical margins, or severe comorbidity, suggesting the importance of early diagnosis, early treatment, negative surgical margins, and conditions of comorbidity.

Supracricoid partial laryngectomy cricohyoidoepiglottopexy (SCPL-CHEP) versus vertical partial laryngectomy for the treatment of glottic carcinoma.

The study aimed to compare the survival rate and functional outcome of supracricoid partial laryngectomy cricohyoidoepiglottopexy (SCPL-CHEP) and vertical partial laryngectomy (VPL) in the treatment of glottic carcinoma. Thirty-nine patients underwent SCPL-CHEP and 45 patients underwent VPL between 2002 and 2007. Post-operative clinical staging were performed according to the UICC staging criteria (2002). The primary endpoints were survival time and recurrence rate. The secondary endpoints were the laryngeal functions including pronunciation, swallowing, and respiration. Overall, the estimated mean survival time was 75.6 months. There was no significant difference in survival times between the two groups (P = 0.496). The SCPL-CHEP group had a significantly lower post-operative local recurrence rate than VPL group (2.6 vs. 17.8 %, P = 0.033). Significantly longer nasogastric tube removal (22 vs. 14 days, P < 0.001) and tracheostomy decannulation (38.5 vs. 15 days, P < 0.001) times in SCPL-CHEP group was observed with the VPL group, respectively. Significantly higher aspiration rates were observed at 3, 6, and 12 months in the SCPL-CHEP group compared with the VPL group (P < 0.001, P < 0.001, P < 0.05, respectively). Although the survival rate was not significantly different between the two groups, the local regional control rate and organ preservation in the supracricoid partial laryngectomy cricohyoidoepiglottopexy group was better than those observed in the vertical partial laryngectomy group.

[Long-term outcome of CO2 laser microlaryngoscopic treatment for laryngeal cancer].

OBJECTIVE: To investigate the long-term outcome of CO2 laser microsurgery for laryngeal cancer. METHODS: Seventy patients with laryngeal cancer were treated with CO2 laser microsurgery. All patients were followed up for at least 36 months (36 - 108 months). RESULTS: During the 36-108 months follow-up, 64 patients were alive, and 6 patients died of recurrence. The total 5-year survival rate was 91.4%, 5-year local control rate was 81.4%, 5-year local recurrence rate was 18.6%, and the neck metastasis rate was 4.3%. All survivals had normal breathing and good phonation. CONCLUSIONS: The long-term outcomes of CO2 laser microsurgery for laryngeal cancer are good, with rapid recovery and few complications, well protected laryngeal function and quite good quality of life. Laser surgery should be the priority of treatment for early stage laryngeal cancer. However, laser surgery for advanced laryngeal cancers and supraglottic laryngeal cancers should be carefully chosen.

[A comparison between endoscopic-assisted submandibular gland resection via retroauricular hairline incision and conventional submandibular gland resection].

OBJECTIVE: To assess the feasibility, the risks and the advantages of endoscope-assisted submandibular gland resection using a retroauricular hairline incision (RAHI) by comparing it with the conventional submandibular gland resection. METHODS: Twenty eight patients with benign lesions of the submandibular gland were included in the prospective clinically controlled study. Thirteen patients had endoscope-assisted resection using the RAHI approach and 15 cases had conventional transcervical approach resection. The size, location and adjacency of all lesions were evaluated by CT or MRI before surgery. The pathologic diagnoses of all cases were identified as benign diseases using fine needle aspiration biopsy. The two groups were compared for incision length, operation time, bleeding, incision cosmetic result, and complications. RESULTS: All 28 operations were successfully performed. Incision length in the endoscopic group was significantly longer than that in the transcervical group (Z = -4.516, P < 0.01), and the surgical time was longer in the endoscopic group (Z = -3.263, P < 0.01). After three months the mean subjective satisfaction score for the incision scar in the endoscopic group was significantly higher than that in the transcervical group (Z = -4.472, P < 0.01). In the endoscopic group, 2 cases (15.4%) with temporary numbness of the earlobe and 1 case (7.7%) with a temporary marginal mandibular nerve paralysis were found postoperatively. However, they recovered within 1 month. All 28 patients were disease free with a follow-up of 10 to 24 months (median of 18 months). CONCLUSIONS: Endoscope-assisted submandibular gland resection via RAHI is feasible and safe for the treatment of benign submandibular gland lesions. In comparison with the transcervical approach, this method can provide better cosmetic results without significant complications.

[Diagnosis and treatment of congenital fourth branchial anomaly].

OBJECTIVE: To discuss the anatomic features, clinical presentations, diagnosis, differentiations and treatments of congenital fourth branchial anomaly(CFBA). METHODS: The clinical data of 8 patients with CFBA were retrospectively analyzed. RESULTS: Of the 8 patients aging from 27 to 300 months (median age: 114 months), 4 male and 4 female; 3 untreated previously and 5 recurrent. All lesions, including 1 cyst, 3 sinus (with internal opening) and 4 fistula, located in the left necks. Three patients presented acute suppurative thyroiditis, 4 deep neck abscesses, and 1 neck lump. Preoperative examinations included barium esophagogram, direct laryngoscopy, ultrasonography, CT, MRI, and so on. The principles of managements were adequate drainage, infection control during acute period and radical surgery during quiescent period. Classic surgical approach consisted of complete excision of branchial lesions, dissection of recurrent laryngeal nerve and partial thyroidectomy. Selective neck dissection was applied in recurrent cases to extirpate branchial lesions, scarrings and inflammatory granuloma. Postoperatively, 1 case was with local incision infection which healed by wound care; 1 case was with temporary vocal cord paralysis which completely recovered 1 month after operation. No recurrence was found in all of 8 cases with follow-up of 13 to 42 months (median: 21 months). CONCLUSIONS: CFBA relates closely anatomically with recurrent laryngeal nerve and thyroid grand. The barium esophagogram and direct laryngoscopy are the most useful diagnostic tools. CT and MRI are all beneficial to the diagnosis of CFBA. The treatment key to CFBA is the complete excision of lesion during a quiescent period after inflammatory control, together with the dissection of recurrent laryngeal nerve, partial thyroidectomy and partial resection of lamina of thyroid cartilage (if necessary), which all can decrease the risk of complications and recurrence. For recurrent cases, selective neck dissection is a safe and effective surgical procedure.

[Ala499Val (C > T) and Lys939Gln (A > C) polymorphisms of the XPC gene: their correlation with male infertility].

OBJECTIVE: The Ala499Val (C > T) and Lys939Gln (A > C) of the XPC gene are two potentially functional nonsynonymous polymorphisms, which affect the rate of DNA repair and might change XPC production and activity. This study aimed to explore the distribution of these two polymorphisms in the Chinese Han population and their relationship with male infertility. METHODS: We genotyped the two polymorphisms of the XPC gene by the PCR-restriction fragment length polymorphism (PCR-RFLP) method in 318 infertile patients and 228 fertile male controls, detected the frequency of the alleles, and analyzed both the individual and the joint contribution of the two polymorphisms to male infertility. RESULTS: For the Ala499Val (C > T) polymorphism, the frequencies of the CC, CT, and TT genotypes were significantly different in distribution between the patients and the controls (P = 0.020). Males with the TT genotype had a lower risk of male infertility than those with the CC genotype (adjusted OR = 0.49, 95% CI: 0.23-0.88), and even lower than those with both CC and CT genotypes (adjusted OR = 0.39, 95% CI: 0.22-0.71). The Lys939Gln (A > C) polymorphism was not related with male infertility. The combined genotype analysis showed that the individuals with 1-4 risk alleles had a significantly higher risk of male infertility (adjusted OR = 2.75, 95% CI = 1.50-5.04) than those with 0 risk allele. CONCLUSION: The Ala499Val (C > T) polymorphism of the XPC gene is correlated with male infertility and may be a potential genetic risk factor for male infertility in the Chinese Han population.

Mapping of cytotoxic T lymphocytes epitopes in E7 antigen of human papillomavirus type 11.

One of the critical steps in the progression to condyloma acuminatum (CA) is the establishment of a persistent human papillomavirus (HPV) infection, majority of HPV type 6 and 11. Cytotoxic T lymphocytes (CTL), which can be induced by the epitope-based peptides in vitro, are thought to be able to recognize and destroy virus-infected cells. In order to screen and identify HLA-A*0201 restricted HPV-11E7 CTL epitopes, five epitope peptides and tetramers were selected including HPV-11E7 7-15 (TLKDIVLDL), 15-23 (LQPPDPVGL), 47-55 (PLTQHYQIL), 81-89 (DLLLGTLNI) and 82-90 (LLLGTLNIV). Human monocyte-derived dendritic cells (DCs) from HLA-A*0201 healthy individuals were pulsed with these peptides to assess the expression of CD83, CD86, HLA-DR and the secretion of IL-12. The ability of peptide-loaded mature DCs to activate autologous T cells was evaluated by analyzing the frequency of specific tetramer(+) CD8(+) T cells using flow cytometry, and the level of IFN-gamma secretion by ELISA. The ability of the epitope-specific CTLs to kill the target cells was also analysed. It was found that the immature DCs could be fully activated by all the five HPV-11E7 peptides and peptide-loaded mature DCs were able to stimulate the epitope-specific T cells in vitro. There was an increased frequency of CD8(+) T cells specific for the E7 7-15 epitope when compared to other four predicted epitopes of HPV-11E7 (P < 0.05). The epitope-specific CTLs for E7 7-15 induced the strongest cytotoxicity to HPV-11E7 expressing cell line at an E:T ratio of 50:1 (P < 0.05). Taken together, these findings demonstrate that E7 7-15 (TLKDIVLDL) is an HLA-A*0201-restricted CTL epitope of HPV type 11. We propose that this epitope could be more helpful in the characterization of HPV control mechanism and be useful for the development of immunotherapeutic approaches for low-risk HPV infectious diseases such as CA.