Natural killer cells: a future star for immunotherapy of head and neck squamous cell carcinoma.

The interplay between immune components and the epithelium plays a crucial role in the development and progression of head and neck squamous cell carcinoma (HNSCC). Natural killer (NK) cells, one of the main tumor-killing immune cell populations, have received increasing attention in HNSCC immunotherapy. In this review, we explore the mechanism underlying the interplay between NK cells and HNSCC. A series of immune evasion strategies utilized by cancer cells restrict HNSCC infiltration of NK cells. Overcoming these limitations can fully exploit the antineoplastic potential of NK cells. We also investigated the tumor-killing efficacy of NK cell-based immunotherapies, immunotherapeutic strategies, and new results from clinical trials. Notably, cetuximab, the most essential component of NK cell-based immunotherapy, inhibits the epidermal growth factor receptor (EGFR) signaling pathway and activates the immune system in conjunction with NK cells, inducing innate effector functions and improving patient prognosis. In addition, we compiled information on other areas for the improvement of patient prognosis using anti-EGFR receptor-based monoclonal antibody drugs and the underlying mechanisms and prognoses of new immunotherapeutic strategies for the treatment of HNSCC.

Counterion Distribution in the Stern Layer on Charged Surfaces.

Counterion adsorption at the solid-liquid interface affects numerous applications. However, the counterion adsorption density in the Stern layer has remained poorly evaluated. Here we report the direct determination of surface charge density at the shear plane between the Stern layer and the diffuse layer. By the Grahame equation extension and streaming current measurements for different solid surfaces in different aqueous electrolytes, we are able to obtain the counterion adsorption density in the Stern layer, which is mainly related to the surface charge density but is less affected by the bulk ion concentration. The charge inversion concentration is further found to be sensitive to the ion type and ion valence rather than to the charged surface, which is attributed to the ionic competitive adsorption and ion-ion correlations. Our findings offer a framework for understanding ion distribution in many physical and chemical processes where the Stern layer is ubiquitous.

Performance Enhancement of Silicone Rubber Using Superhydrophobic Silica Aerogel with Robust Nanonetwork Structure and Outstanding Interfacial Effect.

The application of high-performance rubber nanocomposites has attracted wide attention, but its development is limited by the imbalance of interface and network effects caused by fillers. Herein, an ultrastrong polymer nanocomposite is successfully designed by introducing a superhydrophobic and mesoporous silica aerogel (HSA) as the filler to poly(methyl vinyl phenyl) siloxane (PVMQ), which increased the PVMQ elongation at break (∼690.1%) by ∼9.3 times and the strength at break (∼6.6 MPa) by ∼24.3 times. Furthermore, HSA/PVMQ with a high dynamic storage modulus (G'0) of ∼12.2 MPa and high Payne effect (ΔG') of ∼9.4 MPa is simultaneously achieved, which is equivalent to 2-3 times that of commercial fumed silica reinforced PVMQ. The superior performance is attributed to the filler-rubber interfacial interaction and the robust filler-rubber entanglement network which is observed by scanning electron microscopy. When the HSA-PVMQ entanglement network is subjected to external stress, both the HSA and bound-PVMQ chains are synergistically involved in resisting structural evolution, resulting in the maximized energy dissipation and deformation resistance through the desorption of the polymer chain and the slip/interpenetrating of the exchange hydrogen bond pairs. Hence, highly aggregated nanoporous silica aerogels may soon be widely used in the application of reinforced silicone rubber or other polymers shortly.

Photocorrosion-Based BiOCl Photothermal Materials for Synergistic Solar-Driven Desalination and Photoelectrochemistry Energy Storage and Release.

Solar-driven interfacial evaporation is one of the most promising desalination technologies. However, few studies have effectively combined energy storage with evaporation processes. Here, a novel multifunctional interfacial evaporator, calcium alginate hydrogel/bismuth oxychloride/carbon black (HBiC), is designed, which integrates the characteristics of interfacial evaporation and direct photoelectric conversion. Under illumination, the Bi nanoparticles which were produced by photoetching of BiOCl and its reaction heat are simultaneously used for the heating of water molecules. Meanwhile, part of the solar energy is converted into chemical energy through the photocorrosion reaction and stored in HBiC. At night, Bi NPs undergo autooxidation reaction and an electric current is generated during this process (like a metal-air battery), in which the maximum current density is more than 15 µA cm-2. This scientific design cleverly combines desalination with power generation and provides a new development direction for energy collection and storage.

Molecular characterization based on tumor microenvironment-related signatures for guiding immunotherapy and therapeutic resistance in lung adenocarcinoma.

Background: Although the role of tumor microenvironment in lung adenocarcinoma (LUAD) has been explored in a number of studies, the value of TME-related signatures in immunotherapy has not been comprehensively characterized. Materials and Methods: Consensus clustering was conducted to characterize TME-based molecular subtypes using transcription data of LUAD samples. The biological pathways and immune microenvironment were assessed by CIBERSORT, ESTIMATE, and gene set enrichment analysis. A TME-related risk model was established through the algorithms of least absolute shrinkage and selection operator (Lasso) and stepwise Akaike information criterion (stepAIC). Results: Four TME-based molecular subtypes including C1, C2, C3, and C4 were identified, and they showed distinct overall survival, genomic characteristics, DNA methylation pattern, immune microenvironment, and biological pathways. C1 had the worst prognosis and high tumor proliferation rate. C3 and C4 had higher enrichment of anti-tumor signatures compared to C1 and C2. C4 had evidently low enrichment of epithelial-mesenchymal transition (EMT) signature and tumor proliferation rate. C3 was predicted to be more sensitive to immunotherapy compared with other subtypes. C1 is more sensitive to chemotherapy drugs, including Docetaxel, Vinorelbine and Cisplatin, while C3 is more sensitive to Paclitaxel. A five-gene risk model was constructed, which showed a favorable performance in three independent datasets. Low-risk group showed a longer overall survival, more infiltrated immune cells, and higher response to immunotherapy than high-risk group. Conclusion: This study comprehensively characterized the molecular features of LUAD patients based on TME-related signatures, demonstrating the potential of TME-based signatures in exploring the mechanisms of LUAD development. The TME-related risk model was of clinical value to predict LUAD prognosis and guide immunotherapy.

Dual targets of lethal apoptosis and protective autophagy in liver cancer with periplocymarin elicit a limited therapeutic effect.

Cardiac glycosides (CGs) are candidate anticancer agents that function by increasing [Ca2+]i to induce apoptotic cell death in several types of cancer cells. However, new findings have shown that the anti­cancer effects of CGs involve complex cell­signal transduction mechanisms. Hence, exploring the potential mechanisms of action of CGs may provide insight into their anti­cancer effects and thus aid in the selection of the appropriate CG. Periplocymarin (PPM), which is a cardiac glycoside, is an active ingredient extracted from Cortex periplocae. The role of PPM was evaluated in HepG2 cells and xenografted nude mice. Cell proliferation, real­time ATP rate assays, western blotting, cell apoptosis assays, short interfering RNA transfection, the patch clamp technique, electron microscopy, JC­1 staining, immunofluorescence staining and autophagic flux assays were performed to evaluate the function and regulatory mechanisms of PPM in vitro. The in vivo activity of the PPM was assessed using a mouse xenograft model. The present study demonstrated that PPM synchronously activated lethal apoptosis and protective autophagy in liver cancer, and the initiation of autophagy counteracted the inherent pro­apoptotic capacity and impaired the anti­cancer effects. Specifically, PPM exerted a pro­-apoptotic effect in HepG2 cells and activated macroautophagy by initiation of the AMPK/ULK1 and mTOR signaling pathways. Activation of macroautophagy counteracted the pro­apoptotic effects of PPM, but when it was combined with an autophagy inhibitor, the anti­cancer effects of PPM in mice bearing HepG2 xenografts were observed. Collectively, these results indicated that a self­limiting effect impaired the pro­apoptotic effects of PPM in liver cancer, but when combined with an autophagy inhibitor, it may serve as a novel therapeutic option for the management of liver cancer.

Corrigendum: Exosomes: Potential biomarkers and functions in head and neck squamous cell carcinoma.

[This corrects the article DOI: 10.3389/fmolb.2022.881794.].

Exosomes: Potential Biomarkers and Functions in Head and Neck Squamous Cell Carcinoma.

Head and neck squamous cell carcinoma (HNSCC), originating from the mucosal epithelial cells of the oral cavity, pharynx, and larynx, is a lethal malignancy of the head and neck. Patients with advanced and recurrent HNSCC have poor outcomes due to limited therapeutic options. Exosomes have active roles in the pathophysiology of tumors and are suggested as a potential therapeutic target of HNSCC. Exosomes in HNSCC have been intensively studied for disease activity, tumor staging, immunosuppression, and therapeutic monitoring. In this review, the biological mechanisms and the recent clinical application of exosomes are highlighted to reveal the potential of exosomes as biomarkers and therapeutic targets for HNSCC.

Efficacy of superselective neck dissection (IIA and III) for supraglottic laryngeal cancer with clinically negative neck.

OBJECTIVES: The purpose of this study was to determine the safety and efficacy of superselective neck dissection(SSND) (levels IIA and III) for patients with supraglottic squamous cell carcinoma (SCC) of the larynx and clinically negative (N0) neck. STUDY DESIGN: This was a prospective analysis of consecutive patients. METHODS: A prospective analysis of 60 patients with SCC of the supraglottic larynx and N0 neck who underwent surgical treatment of the primary lesion with simultaneous SSND (levels IIA and III). The incidence of occult metastasis in the lymph nodes, regional recurrence, survival rate, and spinal accessory nerve function were evaluated. RESULTS: Ninety-eight superselective neck dissection (levels IIA and III) procedures were performed for 60 patients. The occult metastasis rate was 20% (12 of 60). Four patients (6.7%) developed regional recurrence, none of them was in level IIB. The 5-year overall, cancer-specific and disease-free survival rate was 82.8%, 87.6%, and 80.7%, respectively. Spinal accessory nerve function was maintained in all patients. CONCLUSION: Superselective neck dissection removing lymph nodes in levels IIA and III was safe and effective for supraglottic SCC of the larynx with clinically negative neck. The spinal accessory nerve function was maintained without compromising clinical outcome.

SLC15A4 Serves as a Novel Prognostic Biomarker and Target for Lung Adenocarcinoma.

BACKGROUND: SLC15A family members are known as electrogenic transporters that take up peptides into cells through the proton-motive force. Accumulating evidence indicates that aberrant expression of SLC15A family members may play crucial roles in tumorigenesis and tumor progression in various cancers, as they participate in tumor metabolism. However, the exact prognostic role of each member of the SLC15A family in human lung cancer has not yet been elucidated. MATERIALS AND METHODS: We investigated the SLC15A family members in lung cancer through accumulated data from TCGA and other available online databases by integrated bioinformatics analysis to reveal the prognostic value, potential clinical application and underlying molecular mechanisms of SLC15A family members in lung cancer. RESULTS: Although all family members exhibited an association with the clinical outcomes of patients with NSCLC, we found that none of them could be used for squamous cell carcinoma of the lung and that SLC15A2 and SLC15A4 could serve as biomarkers for lung adenocarcinoma. In addition, we further investigated SLC15A4-related genes and regulatory networks, revealing its core molecular pathways in lung adenocarcinoma. Moreover, the IHC staining pattern of SLC15A4 in lung adenocarcinoma may help clinicians predict clinical outcomes. CONCLUSION: SLC15A4 could be used as a survival prediction biomarker for lung adenocarcinoma due to its potential role in cell division regulation. However, more studies including large patient cohorts are required to validate the clinical utility of SLC15A4 in lung adenocarcinoma.

Long noncoding RNAs in head and neck squamous cell carcinoma: biological functions and mechanisms.

Head and neck squamous cell carcinoma (HNSCC) is the primary malignant tumor of the oral cavity, larynx, nasopharynx, esophagus and tongue. Although several novel therapeutic methods for HNSCC have been developed, the final therapeutic effect on the patient is still not satisfactory. Thus, it is imperative that scientists identify novel distinguishable markers with specific molecular characteristics that can be used in therapeutic and prognostic evaluation. Previous reports have shown that long noncoding RNAs (lncRNAs) are important regulators of gene expression in many cancers, including head and neck squamous cell carcinomas. Translational studies of lncRNAs in HNSCC are urgently required before their application as a treatment can be realized. We aimed to address the most relevant findings on lncRNAs as biomarkers or treatment targets in head and neck squamous cell carcinoma and to summarize their discovered pathways and mechanisms of action to reveal the possible future applications of these novel biomarkers in clinical translational research.

Long noncoding RNA LINC01194 enhances the malignancy of laryngeal squamous cell carcinoma by sponging miR-655 to increase SOX18 expression.

Long noncoding RNAs (lncRNAs) have undergone a comprehensive study for their involvements in tumor treatments. The purpose of our study was to explore the biological effects and regulatory mechanisms of lncRNA LINC01194 (LINC01194) in laryngeal squamous cell carcinoma (LSCC). The levels of LINC01194 in 105 LSCC patients were detected by RT-qPCR. The diagnostic and prognostic value of LINC01194 in LSCC patients were statistically analyzed. The potential functions of LINC01194 in proliferation, apoptosis, and metastasis of LSCC cells were evaluated. The interaction among LINC01194, miR-655 and SOX18 was explored by bioinformatics analysis, luciferase reporter assays and biotinylated RNA pull-down. We found that the expression levels of LINC01194 were highly expressed in LSCC, which was negatively correlated with the clinical outcome of LSCC patients. The area under the ROC curve for LINC01194 was up to 0.8388. Functional assays indicated that LINC01194 knockdown distinctly inhibited LSCC cells proliferation, induced apoptosis, and also attenuated LSCC cells migration and invasion in vitro. Furthermore, we elucidated that LINC01194 promoted SOX18 expression in LSCC cells via functioning as a molecular sponge for miR-655. Overall, based on our findings, LINC01194 served as a tumor promoter and potentially represents a novel prognostic indicator and therapeutic target in LSCC.

Novel variant strains of infectious bursal disease virus isolated in China.

Infectious bursal disease (IBD) is one of the most important immunosuppressive diseases that seriously threaten poultry farming and food safety worldwide. The variant strain of infectious bursal disease virus (IBDV) has been greatly neglected for more than 30 years. Recently, the subclinical infection of suspected IBD, causing considerable economic losses, occurred in the main chicken-farming regions of China. Through RT-PCR, sequencing, and phylogenic analyses, novel variant IBDVs were first identified in six provinces of eastern China. Immunological detection further confirmed the antigenic variation of the Chinese variant IBDVs. The Chinese IBDV variants were obviously different from the American IBDV variants, with less than a 97.7% (VP1) or 98.7% (VP2) amino acid sequence identity. Animal experiments further confirmed the serious threat of the variant IBDVs to chickens, demonstrating irreversible damage to the central immune organ, obvious immunosuppression, and growth retardation. This study not only identified the pandemic nature of the novel variant IBDVs for the first time but also discovered the distinct molecular epidemiological characteristics of these viruses, which will contribute more to the control of the disease.

LncRNA NORAD promotes thyroid carcinoma progression through targeting miR-202-5p.

Long noncoding RNAs (lncRNAs) have been shown to have several functional roles in tumor biology, and they are deregulated in many types of cancer. The role of a novel lncRNA, NORAD, in papillary thyroid carcinoma (PTC) is still unknown. In this study, we demonstrated that NORAD expression was upregulated in PTC cell lines and samples. Ectopic expression of NORAD promoted PTC cell growth, invasion and migration. Overexpression of NORAD promotes epithelial-to-mesenchymal transition (EMT) progression in the PTC cell. Furthermore, overexpression of NORAD suppressed miR-202-5p expression in PTC cells. The data suggested that miR-202-5p expression was downregulated in PTC cell lines and samples and was negatively correlated with NORAD expression in PTC tissues. Overexpression of miR-202-5p suppressed PTC cell growth, invasion and migration. In addition, we demonstrated that elevated expression of NORAD promoted PTC cell growth, invasion and migration by inhibiting miR-202-5p expression. These results suggested that the lncRNA NORAD acts as an oncogene in PTC progression, partly by regulating miR-202-5p expression.

TSR2 Induces laryngeal cancer cell apoptosis through inhibiting NF-κB signaling pathway.

OBJECTIVES/HYPOTHESIS: Human laryngeal squamous cell carcinoma (LSCC) is a malignancy that was discovered originally in the epithelial tissue of laryngeal mucosa. However, the underlying molecular mechanism is still not clear. In this study, we aimed to investigate the potential molecular mechanisms of TSR2 in the LSCC cell apoptosis. STUDY DESIGN: The expression of TSR2 was first analyzed in LSCC tissues. Then functional effects of TSR2 on Hep-2 and AMC-HN-8 cell lines were performed by overexpression pcDNA3.1-TSR2. METHODS: We investigated the expression level of TSR2 in LSCC tissues and cells by performing quantitative real-time polymerase chain reaction (qRT-PCR). The pcDNA3.1-TSR2 was constructed to explore the effect of overexpressing TSR2 in Hep-2 cells and AMC-HN-8 cells. We further investigated the effect of overexpressing TSR2 on cell apoptosis-related protein and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 nuclear translocation through Western blot and terminal dUTP nick end-labeling assays. RESULTS: We found that TSR2 was downregulated in LSSC tissues and cells compared with the controls, and the overexpression of TSR2 in Hep-2 and AMC-HN-8 cells could promote cell apoptosis and related apoptosis proteins. The Western blot/qRT-PCR data further indicated that overexpression of TSR2 in Hep-2 and AMC-HN-8 cells could lead to a block of NF-κB signaling pathway via decreasing nuclear NF-κB p65 and increasing cytoplasm NF-κB p65. Moreover, overexpression of TSR2 significantly inhibited the phosphorylation of IκBα and IKKα/ß. CONCLUSIONS: The results indicated that TSR2-induced apoptosis was mediated by inhibiting the NF-κB signaling pathway, which may provide an effective target in gene therapy for LSCC. LEVEL OF EVIDENCE: NA. Laryngoscope, 128:E130-E134, 2018.

Krüppel-like factor 5: a novel biomarker for lymph node metastasis and recurrence in supraglottic squamous cell laryngeal carcinoma.

Krüppel-like factor 5 (KLF5), a zinc finger-containing transcription factor, is involved in important biological processes including cell transformation, proliferation, and carcinogenesis. However, its clinical significance has remained largely unknown in laryngeal cancer. Here, specimens from 144 patients with laryngeal tumors were investigated by immunohistochemical staining for KLF5, integrin-linked kinase (ILK), and E-cadherin expressions. A clinicopathological study revealed that the KLF5 expression level in tumor cells was significantly correlated with lymph node metastasis (P < 0.05) and local recurrence (P < 0.05). In addition, KLF5, ILK, and E-cadherin (epithelial-mesenchymal transition (EMT) biomarker) expressions were correlated with each other. These findings suggest that KLF5 may be an epithelial-mesenchymal transition-associated biomarker in human laryngeal carcinomas and play important roles in the progression of laryngeal carcinomas. KLF5 immunoreactivity is therefore considered a potential lymph node metastasis and recurrence factor in human laryngeal cancers. In addition, the KLF5-mediated pathway is a potential target for elimination of laryngeal cancer in the future.

Spironolactone inhibits apoptosis in rat mesangial cells under hyperglycaemic conditions via the Wnt signalling pathway.

Mesangial cells (MCs) play a crucial role in maintaining structure and function of glomerular tufts, providing structural support for capillary loops and modulating glomerular filtration by their contractility. MCs apoptosis occurs in experimental diabetic nephropathy, and this correlates with worsening albuminuria. Accumulating evidence suggests that mineralocorticoid receptor (MR) blockade effectively reduces proteinuria in diabetic nephropathy; however, it is rarely known whether spironolactone (SPI), a nonspecific MR antagonist, inhibits apoptosis in MCs under hyperglycaemic conditions. The objectives of this study are to determine the relationship between SPI and apoptosis, and investigate the cell signalling pathway by which SPI inhibits apoptosis. Rat MCs were treated with 30 mM D-glucose and 10(-8), 10(-7) or 10(-6) M aldosterone (ALD) for 24 h. In some experiments, MCs were pretreated with 10(-7) M SPI or 10 mM LiCl for 1 h. Apoptosis was evaluated by cell nucleus staining and flow cytometric analyses, and caspase-3 activity was assayed. Gene and protein expression were quantified using quantitative real-time PCR and Western blotting, respectively. SPI directly inhibited high glucose and ALD-induced MCs apoptosis in a caspase-dependent manner. Importantly, SPI inhibited MCs apoptosis via the Wnt signalling pathway. SPI promoted activation of the Wnt signalling pathway in MCs, leading to upregulation of Wnt4 and Wnt5a mRNA expression, decreased GSK-3ß protein expression and increased ß-catenin protein expression. As a conclusion, this study suggests that SPI may inhibit apoptosis in MCs during hyperglycaemic conditions via the Wnt signalling pathway. Blockade of the ALD system may represent a novel therapeutic strategy to prevent MCs injury under hyperglycaemic conditions.

Krüppel-like factor 8 is a novel androgen receptor co-activator in human prostate cancer.

AIM: Krüppel-like factor 8 (KLF8) plays important roles in cell cycle and oncogenic transformation. On other hand, androgen receptor (AR) is crucial in development of both androgen-dependent and independent prostatic malignancies. The aim of this study is to investigate the role of KLF8 in prostate cancer (PCa) and the relationship between KLF8 and AR. METHODS: Eight human PCa cell lines, including androgen-dependent LNCap cells and androgen-independent 22Rv1 cells, as well as human PCa samples were studied. LNCap cells and 22Rv1 cells were transfected with plasmids encoding full-length wild-type KLF8 or KLF8 shRNA. The expression of KLF8 protein was detected using Western blotting or immunohistochemical staining. Cell proliferation in vitro was measured with MTT assay, and in vivo in a xenograft nude mouse model. Yeast two-hybrid screening, co-immunoprecipitation and pull down assays were used to examine the binding of KLF8 to AR. Luciferase reporter gene assay was used to measure the transcriptional activity of the genes targeted by AR. RESULTS: In 133 human PCa samples, KLF8 protein staining was observed in 92.65% (63/68) of high-grade PCa, 66.15% (43/65) of low-grade PCa, and 6.82% (3/44) of adjacent normal tissues. The expression of KLF8 was significantly associated with poorer overall survival. Overexpression of KLF8 enhanced the proliferation of both LNCap and 22Rv1 cells, while knockdown of endogenous KLF8 suppressed the proliferation. These manipulations exerted similar effects on the tumor volumes in the xenograft nude mouse model. Yeast two-hybrid screening revealed that KLF8 was a novel AR-interacting protein. With pull down assay and co-immunoprecipitation assay, we demonstrated that KLF8 bound directly to AR, and KLF8 enhanced AR target gene transcription. CONCLUSION: The results demonstrate that KLF8 is a novel AR transcriptional co-activator that is overexpressed in PCa and may play a role in progression of hormone-refractory PCa.

Incidence of level IIB lymph node metastasis in supraglottic laryngeal squamous cell carcinoma with clinically negative neck--a prospective study.

BACKGROUND: The purpose of this study was to determine the incidence of level IIB metastasis in patients with clinically negative (N0) necks with squamous cell carcinoma (SCC) of the supraglottic larynx and to assess whether level IIB lymph nodes may be left undissected in such patients. This was a prospective analysis of a case series. METHODS: A prospective analysis of 68 patients with SCC of the supraglottic larynx and N0 neck who underwent surgical treatment of the primary lesion with simultaneous selective neck dissection (SND) of levels II and III. During neck dissection, level IIB lymph nodes were separately removed and processed. Occult metastasis at levels IIA, IIB, and III and the regional recurrence within this area were evaluated. RESULTS: One hundred twenty-two SND (levels II and III) procedures were performed on 68 patients in this series. The occult metastasis rate was 30.9% (21 of 68). The mean number of lymph nodes harvested in level IIB was 4.1 (range, 1-14). In none of the 122 SND specimens (levels II and III) was metastasis found in level IIB either in the ipsilateral or contralateral neck sides. Five patients developed neck recurrence, none of which was in level IIB. CONCLUSION: When SND was carried out for patients with SCC of the supraglottic larynx with N0 neck, superselective neck dissection removing lymph nodes in levels IIA and III was adequate. Level IIB lymph node pads may be left undissected so that spinal accessory nerve dysfunction is decreased and operative time reduced.

[Metastatic rule and management of occult lymphatic metastasis in supraglottic cancer with clinically negative neck].

OBJECTIVE: To investigate the metastatic rule and management methods of occult lymphatic metastasis in patients with supraglottic cancer. METHOD: The results of 139 cN0 patients with supraglottic cancer treated by neck dissection (ND) were reviewed retrospectively. Each lymph node was studied pathologically in order to define the level of the lymph node metastasis. RESULT: Of the 139 patients with supraglottic cancer and cN0 neck, 113 cases received ipsilateral ND and 26 cases with bilateral ND. Thirty-six (25.9%) of 139 patients were found to have occult lymphatic metastasis on pathological examination, 6 patients without metastasis in dissected side at pathologic examination showed metastasis in the contralateral undissected neck later. Therefore the total occult metastasis rate was 30.2% (42 of 139). The unilateral and bilateral neck occult metastases were determined in 37 cases (26.6%) and 5 cases (3.6%) respectively. A total of 3594 lymph nodes were harvested in 165 specimens, with 83 positive nodes. The distributions of the 83 positive nodes were as follows: level I 1.2% (1 node), level II 78.3% (65 nodes), level III 19.3% (16 nodes), level IV 1.2% (1 node), level V (0 node). The 3-year regional recurrence rate was 5.0% (7/139). The 5-year survival rate was 76.3% (106/139). Patients with positive neck metastasis had higher regional recurrence rates (P < 0.05). CONCLUSION: Occult metastasis rate of supraglottic cancer is as high as 30.2%, the selective lateral neck dissection of level II, III is recommended.