An immune-related lncRNA pairs signature to identify the prognosis and predict the immune landscape of laryngeal squamous cell carcinoma.

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) is the most common squamous cell carcinoma. Though significant effort has been focused on molecular pathogenesis, development, and recurrence of LSCC, little is known about its relationship with the immune-related long non-coding RNA (lncRNA) pairs. METHODS: After obtaining the transcriptome profiling data sets and the corresponding clinical characteristics of LSCC patients and normal samples from The Cancer Genome Atlas (TCGA) database, a series of bioinformatic analysis was conducted to select the differently expressed immune-related lncRNAs and build a signature of immune-related lncRNA pairs. Then, the effectiveness of the signature was validated. RESULTS: A total of 111 LSCC patients and 12 normal samples' transcriptome profiling data sets were retrieved from TCGA. 301 differently expressed immune-related lncRNAs were identified and 35,225 lncRNA pairs were established. After univariate Cox analysis, LASSO regression and multivariate Cox analysis, 7 lncRNA pairs were eventually selected to construct a signature. The riskscore was computed using the following formula: Riskscore = 0.95 × (AL133330.1|AC132872.3) + (-1.23) × (LINC01094|LINC02154) + 0.65 × (LINC02575|AC122685.1) + (-1.15) × (MIR9-3HG|LINC01748) + 1.45 × (AC092687.3|SNHG12) + (-0.87) × (AC090204.1|AL158166.1) + 0.64 × (LINC01063|Z82243.1). Patients were classified into the high-risk group (> 1.366) and the low-risk group (< 1.366) according to the cutoff value (1.366), which is based on the 5-year riskscore ROC curve. The survival analysis showed that the low-risk group had a better prognosis (P < 0.001). The riskscore was better than other clinical characteristics in prognostic prediction and the area under the curves (AUCs) for the 1-, 3-, and 5-year survivals were 0.796, 0.946, and 0.895, respectively. Combining age, gender, grade, stage, and riskscore, a nomograph was developed to predict survival probability in LSCC patients. Then, the riskscore was confirmed to be related with the content of tumor-infiltration immune cells and the model could serve as a potential predictor for chemosensitivity. CONCLUSION: We successfully established a more stable signature of 7 immune-related lncRNA pairs, which has demonstrated a better prognostic ability for LSCC patients and may assist clinicians to precisely prescribe chemo drugs.

Characteristics and clinical significance of polyploid giant cancer cells in laryngeal carcinoma.

OBJECTIVES: We aimed to construct an induction system for polyploid giant cancer cells (PGCCs), as well as to investigate PGCC features and clinical significance. METHODS: A laryngeal neoplasm-PGCC induction system was constructed using paclitaxel liposomes (PTX). We used western blots to compare expression of epithelial-mesenchymal transition-related proteins, stem cell interrelated proteins, and cyclin-associated proteins. We then measured PGCC count in tissue samples of patients with laryngeal neoplasms and analyzed its relationship with prognosis. Statistical significance was determined using t-tests. RESULTS: PTX successfully induced PGCCs. Western blotting showed that CyclinB1, CDC25C, CDK1, E-cadherin, and EIF-4A expression decreased in PGCCs compared with normal cancer cells, whereas vimentin and CD133 expression increased. Number of PGCCs in laryngeal cancer tissues and overall survival time were inversely correlated (P < .05). CONCLUSIONS: PTX successfully induces PGCC formation in laryngeal carcinoma, which may be the cause of poor prognosis in patients with laryngeal cancer.Level of Evidence: 4.

Mining database for the expression and clinical significance of STAT family in head and neck squamous cell carcinomas.

BACKGROUND: Head and neck squamous cell carcinomas (HNSC) are among the most common malignant tumors with high incidence, relapse, and mortality rate. STAT proteins are implicated in various biological processes, including cell proliferation, metastasis, and immune regulation. METHOD: Various bioinformatics tools were used to explore the role of the STAT family in HNSC. RESULT: The mRNA levels of STAT1/2/4/5A/6 were significantly upregulated in HNSC tissues. The levels of STAT1/2/4/5A/6 could be used for the detection of HNSC. HNSC patients with a high level of STAT5A had a poor overall survival and relapse-free survival. A moderate to high correlation was obtained between the STAT family and HNSC. Genetic alteration revealed that STAT1/2/3/4/5A/5B/6 were altered in 6%, 5%, 7%, 8%, 6%, 6%, and 4% of the queried TCGA HNSC samples, respectively. Immune infiltrations analysis suggested a significant association between STAT5A expression and abundance of specific immune cells. Further, copy number alteration of STAT5A could certainly inhibit infiltration level. Moreover, a close correlation was obtained between STAT5A level and the expression of immune markers in HNSC. Several kinase targets and transcription factor targets of STAT5A in HNSC were also identified. Enrichment analysis suggested that STAT5A and co-expression genes were mainly responsible for adaptive immune response, T cell activation, cytokine-cytokine receptor interaction, chemokine signaling pathway, cell-adhesion molecules, and ribosome and RNA transport. CONCLUSION: Our results provided additional data for the expression and clinical significance of the STAT family in HNSC, and further study should be performed to verify these.

Therapeutic efficacy and safety of radiofrequency ablation for the treatment of trigeminal neuralgia: a systematic review and meta-analysis.

OBJECTIVE: The objective of this study was to summarize the effectiveness and safety of trigeminal neuralgia (TN) treatment via different radiofrequency approaches such as continuous radiofrequency (CRF), pulsed radiofrequency (PRF), and combined CRF and pulsed radiofrequency (CCPRF) treatments, thus providing high-quality clinical evidence for TN treatment. METHODS: A series of databases were searched for relevant articles published between January 1998 and April 2018. The modified Jadad scale was referred to evaluate the methodological quality of the included studies. Data were extracted independently, and the outcome and safety of different routes, temperatures, and guidance used in CRF, PRF, and CCPRF were compared. Meta-analysis and publication bias were calculated using Review Manager software. RESULTS: In total, 34 studies involving 3,558 participants were included. With regard to TN treatment, PRF had no difference in cured rate in comparison with CRF, while CRF was more effective than CCPRF (P<0.05). The comparison of complication rates showed that PRF and CCPRF were safer. For puncture guidance via CRF, three-dimensional-printed template was more accurate in success rate at first puncture than computed tomography guidance (P<0.05). For puncture route, foramen rotundum (FR) or pterygopalatine fossa (PPF) route had no significance in efficiency rate via CRF in comparison with foramen oval (FO) route, but PPF and FR routes were safer. For CRF treatment, low temperature (68°C-70°C) compared with high temperature (71°C-75°C) had no effect. Moreover, higher temperature (66°C-80°C) had a greater effect compared with lower temperature (55°C-65°C) on TN treatment (P<0.05), while the safety of which was decreasing. CONCLUSION: CCPRF could achieve a greater effect and safety on TN treatment. FR and FO routes in TN puncture treatment via CRF are safer. Medium temperature range is better for CRF therapy, and higher temperature is recommended in PRF, especially for the elders. Further international multicenter trials are needed to confirm the evidence.

Tra2ß silencing suppresses cell proliferation in laryngeal squamous cell carcinoma via inhibiting PI3K/AKT signaling.

OBJECTIVES/HYPOTHESIS: Transformer-2 protein homolog beta (Tra2ß) generally plays an important role in various human cancers, but its role and the underlying mechanisms in laryngeal squamous cell carcinoma (LSCC) remained unknown. So this study aimed to assess the clinical significance and regulatory mechanisms of Tra2ß in LSCC. STUDY DESIGN: Laboratory analysis. METHODS: Expression of Tra2ß was compared in human LSCC tissue samples and paired adjacent normal tissue samples. The in vitro effects of Tra2ß expression in Hep-2 cells on their proliferation, invasion, and migration were assessed by CCK-8 assays, Matrigel invasion, and transwell migration assays. In addition, the effects of downregulation of Tra2ß on the activation of PI3K/AKT signaling pathway were measured using Western blot analysis. The effect of Tra2ß on the growth of tumors was detected in the Hep-2-injected xenograft models in vivo. RESULTS: Reverse-transcription quantitative polymerase chain reaction analysis and immunochemistry analysis indicated that the increased expression of Tra2ß in LSCC was significantly associated with poor differentiation, lymph node metastasis, and advanced clinical stage. In vitro knockdown of Tra2ß caused a significant decrease in the proliferation, invasion, and migration of Hep-2 cells. Tra2ß silencing decreased the expression of Bcl-2 but increased Bax and Caspase-3 both in mRNA and protein levels. Furthermore, knockdown of Tra2ß eliminated the suppressive effects of activation of PI3K/AKT signaling. In vivo knockdown of Tra2ß significantly inhibited the tumor growth of Hep-2-injected xenograft mice. CONCLUSIONS: The results of the present study demonstrated that knockdown of Tra2ß inhibits the proliferation and invasion of LSCC cells, at least partly via inhibiting PI3K/AKT signaling. LEVEL OF EVIDENCE: NA Laryngoscope, 129:E318-E328, 2019.

Spondin-2 is a novel diagnostic biomarker for laryngeal squamous cell carcinoma.

Spondin-2, belongs to the SOX (SRY-related HMG box) gene family, plays a vital role in the development of malignancy, however, the role of Spondin-2 in laryngeal squamous cell carcinoma (LSCC) remains unknown. The aim of this study is to investigate the prognostic significance of and probable mechanism of Spondin-2 in LSCC. qRT-PCR, western blotting assays and IHC analysis demonstrated that Spondin-2 was significantly increased in LSCC tissues compared with adjacent non-tumorous tissues. In addition, high levels of Spondin-2 was associated with clinical stage, lymph node metastasis and pathology grade of LSCC patients (P ＜0.05). Kaplan-Meier analysis showed that patients with high expression of Spondin-2 had a lower overall survival rate (P＜0.05) than that with low expression of Spondin-2. Moreover, spondin-2 silencing inhibited the proliferation of LSCC cells through inhibiting the activation of PI3K/AKT signaling. In conclusion, spondin-2 might be a novel therapeutic target and prognostic biomarker for LSCC patients.

Elevated EGFL6 modulates cell metastasis and growth via AKT pathway in nasopharyngeal carcinoma.

Epidermal growth factor-like domain multiple 6 (EGFL6) is a secreted protein, regulates maintenance and metastasis of cancer cells. Nevertheless, how EGFL6 participates in the progression and tumorigenesis of nasopharyngeal carcinoma (NPC) remains unclear. In our study, EGFL6 was detected highly expressed in 20 NPC tissues compared with normal tissues by IHC assay. Then, the level of EGFL6 in NPC serum and NPC cells was explored through enzyme-linked immunosorbent assay and western blot, the results consistent with IHC. More interestingly, EGFL6 accelerated the migration and growth of NPC in vitro assays. Considering the mechanism of migration, NPC cells were cultured with AKT activator, revealing EGFL6 facilitated the progression of NPC via AKT. Moreover, the same effect of EGFL6 in promoting NPC growth was proved in nude mice. Furthermore, heat-shock zebrafish model was established with EGFL6 overexpression. Then, CNE2 cells were injected into the model and cells mass was observed, showing that EGFL6 enhanced the migration and metastasis of NPC. Currently, as the prognosis of NPC is severely affected by distant metastasis, it might be a new therapeutic target toward EGFL6. Taken together, our results suggested that EGFL6 acts as a potential positive regulator in the migration and proliferation of NPC.

A disintegrin and metalloprotease 10-containing exosomes derived from nasal polyps promote angiogenesis and vascular permeability.

Abnormal angiogenesis and vascular permeability is important for the formation of nasal polyps (NPs). Increasing evidence has indicated that exosomes serve a vital role in modulating angiogenesis and vascular permeability. A disintegrin and metalloprotease 10 (ADAM10), an important type of proteinase that is overexpressed in various diseases, can influence angiogenesis and vascular permeability and has been observed in healthy nasal exosomes. To the best of our knowledge, the expression levels and the function of ADAM10 in NLF­derived exosomes from NPs has not been demonstrated previously. In order to determine the influence of exosomes derived from nasal lavage fluid (NLF) on angiogenesis and vascular permeability, 25 nasal polyp patients and 15 healthy volunteers were enrolled in the present study. NLF was collected from all of the subjects. Exosomes were isolated from NLF, visualized under transmission electron microscope and identified using western blot analysis. The effect of exosomes on human umbilical vein endothelial cells (HUVECs) was measured by tube formation and permeability assays in vitro. The expression of exosomal ADAM10 was also analyzed by western blotting. NLF­derived exosomes from NPs influenced proliferation, tube formation and the permeability of HUVECs. ADAM10 was highly expressed in NLF­derived exosomes from NPs when compared with healthy volunteers. Thus, NLF­derived exosomes from NPs promoted angiogenesis and vascular permeability, which may be associated with abundant ADAM10 in NP exosomes.

FoxM1 overexpression promotes cell proliferation and migration and inhibits apoptosis in hypopharyngeal squamous cell carcinoma resulting in poor clinical prognosis.

Forkhead box M1 (FoxM1), a member of the Fox family of transcriptional factors, is involved in the development of various human malignancies. However, the expression level of FoxM1 and its functional role in hypopharyngeal squamous cell carcinoma (HSCC) remained unclear to date. The aim of the present study was to investigate the FoxM1 expression in 63 HSCC and 20 adjacent normal tissues, as well as to evaluate its association with the clinicopathological parameters and its diagnostic value in HSCC. To further explore the biological function of FoxM1 in vitro, siRNAs were used to knockdown the expression of FoxM1 in the HSCC cell line Fadu. The results revealed that FoxM1 protein was highly expressed in HSCC tissues and that its high expression was closely associated with HSCC tumor differentiation (P=0.004), tumor size (P=0.002), clinical stage (P=0.001), lymph node metastasis (P=0.002), treatment (P=0.045) and expression of the proliferation marker Ki-67 (P<0.001). Additionally, the elevated expression of FoxM1 in HSCC patients consistently predicted a poor survival time. Knockdown of FoxM1 expression blocked Fadu cell proliferation and promoted apoptosis, and also led to the downregulation of cyclin A1 expression. Furthermore, decreased expression of FoxM1 markedly impeded cell migration and reversed the epithelial-mesenchymal transition phenotype, as indicated by decreased expression of vimentin and increased expression of E-cadherin in Fadu cells. These results indicate that FoxM1 may act as an oncogene and serve as a therapeutic target against malignant progression in HSCC.

Clinical and biological significance of HAX-1 overexpression in nasopharyngeal carcinoma.

HS1-associated protein X-1 (HAX-1) is an important marker in many types of cancers and contributes to cancer progression and metastasis. We examined the expression of HAX-1 in nasopharyngeal carcinoma (NPC) and experimentally manipulated its expression. We observed that HAX-1 expression is elevated in NPC and is correlated with lymph node metastasis, M classification, clinical stage, and poor prognosis. In addition, overexpression of HAX-1 promoted NPC proliferation both in vitro and in vivo. Exosomes are potential carriers of pro-tumorigenic factors that participate in oncogenesis. We found that NPC-derived exosomes are enriched in HAX-1 and accelerate NPC tumor growth and angiogenesis in vitro and in vivo. Furthermore, we demonstrated that oncogenic HAX-1 facilitates the growth of NPC when it is transferred via exosomes to recipient human umbilical vein endothelial cells (HUVECs). Oncogenic HAX-1 also increases the proliferation, migration, and angiogenic activity of HUVECs. Our findings provide unique insight into the pathogenesis of NPC and underscore the need to explore novel therapeutic targets such as HAX-1 to improve NPC treatment.

[Research on the efficacy and mechanism of standardized house dust mite allergen vaccine to allergic rhinitis].

OBJECTIVE: The study was designed to analyze the efficacy of standardized house dust mite allergen vaccine to allergic rhinitis by subcutaneous injection and investigate the possible mechanism of specific immunotherapy (SIT). METHODS: From January 2011 to December 2011 a prospective study was performed in the Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Nantong University, involving 90 patients with perennial AR, of whom 60 patients received Der p - SIT + pharmacotherapy after their approval and 30 received only pharmacotherapy. All patients were allergic to house dust mites. Symptom and medication scores were recorded three times: before the treatment, at the middle of treatment and at the end of treatment. Over a period of 1 yr. prior to and at the end of treatment, CD4⁺ CD25⁺ Foxp3⁺ Treg cells and Th17 cells were measured by flow cytometry. SPSS 21.0 software was used to analyze the data. RESULTS: The symptom scores using VAS and medication scores in AR patients treated with SIT and medication were reduced, the differences were significant (14.25 ± 6.40, 1.00 ± 0.84 vs. 32.18 ± 7.78, 3.12 ± 1.54, t value was 19.65, 10.71, both P < 0.05). The symptom of VAS score in medication group was reduced after treatment (30.30 ± 5.97 vs. 20.30 ± 5.79, t = 10.09, P < 0.05), but the medication score had not significant difference (P > 0.05). The frequency of Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT, whereas the frequency of Treg cells were increased (χ² value was 2.81, 2.80, both P < 0.05), but not in medication group. CONCLUSIONS: Both SIT and pharmacotherapy can improve symptoms of allergic rhinitis, but SIT can also reduce medication use. The effect of immunotherapy is better than drug treatment alone. The frequency of blood Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT, whereas the frequency of Treg cells were increased.

Clinical significance of HAX-1 expression in laryngeal carcinoma.

OBJECTIVE: HS1-associated protein X-1 (HAX-1) is a multifunctional protein that has been highlighted as an important marker in many types of cancers. However, little is known about the role of HAX-1 in laryngeal carcinoma. The purpose of the present study is to explore HAX-1 expression status and its associations with clinicopathologic features and survival in a well-defined cohort of laryngeal carcinoma. METHODS: We examined the expression of HAX-1 at protein and mRNA levels in laryngeal carcinoma tissues and adjacent non-tumor tissues by immunohistochemistry, Western blotting and two-step quantitative real-time PCR analysis, respectively. RESULTS: We observed that HAX-1 was significantly elevated in laryngeal carcinoma. The relationship between the levels of HAX-1 expression and clinicopathologic characteristics was then analyzed. Overexpression of HAX-1 was significantly correlated with T classification, lymph node metastasis, clinical stage, and pathology. Survival curves were plotted using the Kaplan-Meier method and compared using the log-rank test. We find that patients with overexpression of HAX-1 had shorter overall survival rates. Finally, the significance of various survival variables was analyzed using multivariate Cox proportional hazards model. We found that overexpression of HAX-1 was an independent prognostic factor for patients with laryngeal carcinoma. CONCLUSION: Our findings hinted that overexpression of HAX-1 was a potentially unfavorable factor in the progression and prognosis of laryngeal carcinoma.

Expression and significance of angiopoietin-2 and cyclin D1 in laryngeal squamous cell carcinoma and the correlation with prognosis.

The aim of this study was to study the expression of angiopoietin-2 (Ang-2) and the cell cycle protein D1 (cyclin D1) in laryngeal squamous cell carcinoma (SCC), and its clinicopathological meaning. The expression of Ang-2 and cyclin D1 was analyzed in 116 cases of laryngeal SCC, five cases of atypical hyperplasia and five cases of vocal polyp tissues using the immunohistochemical streptavidin-perosidase (S-P) method. Ang-2 and cyclin D1 protein expression was not present in 5 cases of atypical hyperplasia or 5 cases of vocal cord polyps. However, the proportions of positive staining in well, moderately and poorly differentiated laryngeal SCC were 40, 66.7 and 100%, respectively, for Ang-2 and 50, 66.7, 100%, respectively, for cyclin D1, and were statistically significant (P<0.05). The expression of Ang-2 was positively correlated with cyclin D1 in the laryngeal SCC (r=0.5042; P<0.05). This showed that the tumor grading and cyclin D1 were independent factors affecting laryngeal SCC patient survival by the Cox regression model of risk factors proportion analysis. Ang-2 is synergistic with cyclin D1 in the development of laryngeal SCC. Ang-2 is associated with the metastasis of lymph nodes. Detection of both Ang-2 and cyclin D1 may possess clinical significance in evaluating the prognosis and guiding the clinical treatment of SCC.