OBJECTIVE: To explore the association of XRCC3 gene polymorphisms and haplotypes with laryngeal. METHOD: We selected 4 tag SNPs (rs12432907, rs861536, rs861537, rs861531, rs861531) for the present study. 310 laryngeal patients and 310 healthy control subject were genotyping. The distribution of genotypes and haplotypes in these two group was compared. RESULT: The distributions of rs12432907 was significantly different between these two groups. The CCAG haplotype frequency was higher in laryngeal group than that in control group. But TCAG and TTAG haplotype frequency was were lower in the laryngeal patient than that those in the control subject. CONCLUSION XRCC3 gene polymorphism was associated with the risk of laryngeal patients.
Case-Control Studies ; DNA-Binding Proteins ; genetics ; Genotype ; Haplotypes ; Humans ; Laryngeal Neoplasms ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVE: To observe expression of stathmin gene in laryngeal squamous cell carcinoma (LSCC) and relation between expression of stathmin gene and occurrence and development of LSCC. METHOD: The expression of the stathmin gene was determined in 35 LSCC of specimens and 18 normal laryngeal tissues (NLT) of specimens by in situ hybridization with Digoxigenin labeled probe of stathmin mRNA. RESULT: Expression of stathmin gene was observed in 35 cases of laryngeal squamous cell carcinoma tissue (positive rate, 69%) and positive signal was observed in both cytoplasm and nuclear. Among 18 cases of normal tissue, only 6 showed weak positive signal. There was significant difference in expression of stathmin gene between laryngeal squamous cell carcinoma tissue and normal tissue. CONCLUSION Expression of stathmin gene may play a key role in the pathogenesis and development of laryngeal squamous cell carcinoma. It may be a very important biotherapy target in the treatment of laryngeal squamous cell carcinoma.
Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Gene Expression ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; RNA, Messenger ; genetics ; Stathmin ; genetics ; metabolism

Carcinoma, Squamous Cell ; genetics ; Cell Division ; Chromosome Deletion ; Humans ; Laryngeal Neoplasms ; genetics ; Telomere ; genetics ; Tumor Cells, Cultured

OBJECTIVE: To study the impacts of RelA antisense oligonucleotides on proliferation in laryngeal carcinoma Hep-2 cell. METHOD: RelA antisense oligonucleotides was designed, which was transferred into laryngeal carcinoma Hep-2 cell. MTT was used to detect the growth-inhibiting ratio at different transferred timepoints. Hep-2 cell which was transferred 48 h was used to do colony assay, and expression of RelA was detected by Reverse Transcription PCR and Western blot. RESULT: MTT results showed that RelA antisense oligonucleotides could significantly suppress the proliferation of Hep-2 cell, and the suppression-ratio elevated with time. There were statistical difference compared with control groups. The number of cells colony was reduced in RelA antisense oligonucleotides group compared with control groups, which had statistic significance. RT-PCR and Western blot results demonstrated that RelA antisense oligonucleotides could significantly inhibit the expression of messenger RNA and protein in Hep-2 cell. CONCLUSION RelA antisense oligonucleotides can inhibit the expression of messenger RNA and protein, and induce the cell proliferation and increase the number of cells colony in Hep-2 cell.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Oligonucleotides, Antisense ; genetics ; pharmacology ; Transcription Factor RelA ; genetics

OBJECTIVE: To detect the expression of microRNAs (miRNAs) in laryngeal carcinoma and adjacent normal tissue using microarray, and to discuss the relationship between miRNAs and laryngeal carcinoma. METHOD: miRNA were extracted from 8 cases of laryngeal cancer tissue and its adjacent normal tissue. miRNA identification were performed by microarray of miRNA hybridization and cluster analysis was used with Significance Analysis of Microarrays (SAM, version 2.1) and Cluster 3.0 software. miRNA were confirmed by real time quantification RT-PCR with RNA-tailing and primer extension. RESULT: Totally 47 different miRNAs were found expressed in laryngeal cancer, with 23 of miRNA expression were up-regulated and 24 of miRNA expression were down-regulated. The expression of miR-1, miR-486-5p, miR-206, miR-487a,miR-375, miR-422a, miR-144, miR-384, miR-378, miR-133a were down-regulated by 5 multiple and while expression of miR-93, miR-31, miR-20b were up-regulated by 3 multiple. There are 5 miRNA clusters with coexpression in laryngeal cancer tissue and located on chromosome 8, 13, 14, 18 and X. Moreover, RT-PCR analysis demonstrated that there was no significantly difference of miRNA expression between microarray and RT-PCR. CONCLUSION The different expression of miRNA may play an important role in the pathogenesis and development of laryngeal cancer.
Adult ; Female ; Gene Expression Profiling ; Humans ; Laryngeal Mucosa ; pathology ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; MicroRNAs ; genetics ; Microarray Analysis ; Middle Aged ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo search for characteristic chromosome changes in primary laryngeal squamous cell carcinoma (LSCC) and Hep-2 cell line and to realize the relationship between the cytogenetic abnormality and the pathogenetic mechanism in LSCC.

METHODSThe fresh resulted samples of LSCC were analyzed with an improved primary cell culture for chromosome preparation and G-banding technique. Hep-2 cell line was analyzed by high resolution banding technique. Molecular cytogenetics analysis was made by chromosome 6 painting probe.

RESULTSFour primary LSCC succeeded in primary cell culture and obtained metaphases, one was tetraploid, the other three were triploid. The chromosome mode of Hep-2 cell line was from 68 to 75 and fifteen marker chromosomes were found. The most structural abnormalities of chromosome in primary LSCC and HEP-2 cell line were unbalance translocation, terminal deletion and isochromosome. The complicate aberration in chromosome 6 was common in LSCC and Hep-2.

CONCLUSION6q-, I(5p), 17p-, 5q- are considered as characteristic chomosome changs in LSCC. Fluorescence in situ hybridization (FISH) may enhance the ability of detecting complicated chromosome rearrangements and marker chromosomes, which could provide more value data to verify the chromosome characteristic aberration in LSCC.

OBJECTIVES: Laryngeal cancer (LC) is a globally prevalent and highly lethal tumor. Despite extensive efforts, the underlying mechanisms of LC remain inadequately understood. This study aims to conduct an innovative bioinformatic analysis to identify hub genes that could potentially serve as biomarkers or therapeutic targets in LC. METHODS: We acquired a dataset consisting of 117 LC patient samples, 16 746 LC gene RNA sequencing data points, and 9 clinical features from the Cancer Genome Atlas (TCGA) database in the United States. We employed weighted gene co-expression network analysis (WGCNA) to construct multiple co-expression gene modules. Subsequently, we assessed the correlations between these co-expression modules and clinical features to validate their associations. We also explored the interplay between modules to identify pivotal genes within disease pathways. Finally, we used the Kaplan-Meier plotter to validate the correlation between enriched genes and LC prognosis. RESULTS: WGCNA analysis led to the creation of a total of 16 co-expression gene modules related to LC. Four of these modules (designated as the yellow, magenta, black, and brown modules) exhibited significant correlations with 3 clinical features: The age of initial pathological diagnosis, cancer status, and pathological N stage. Specifically, the yellow and magenta gene modules displayed negative correlations with the age of pathological diagnosis (r=-0.23, P<0.05; r=-0.33, P<0.05), while the black and brown gene modules demonstrated negative associations with cancer status (r=-0.39, P<0.05; r=-0.50, P<0.05). The brown gene module displayed a positive correlation with pathological N stage. Gene Ontology (GO) enrichment analysis identified 77 items, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 30 related signaling pathways, including the calcium signaling pathway, cytokine-cytokine receptor interaction, neuro active ligand-receptor interaction, and regulation of lipolysis in adipocytes, etc. Consequently, central genes within these modules that were significantly linked to the overall survival rate of LC patients were identified. Central genes included CHRNB4, FOXL2, KCNG1, LOC440173, ADAMTS15, BMP2, FAP, and KIAA1644. CONCLUSIONS This study, utilizing WGCNA and subsequent validation, pinpointed 8 genes with potential as gene biomarkers for LC. These findings offer valuable references for the clinical diagnosis, prognosis, and treatment of LC.
Humans ; Laryngeal Neoplasms/genetics* ; Rosaniline Dyes ; Biomarkers ; Adipocytes ; Gene Regulatory Networks ; Gene Expression Profiling

The relationship between hypermethylation of CpG islands in the promoter regions of O6-methylguanine DNA methyltransferase (MGMT) genes and laryngeal squamous cell carcinoma was explored. Methylation-specific PCR and semi-quantitative RT-PCR were used to study the promoter methylation and mRNA expression of the MGMT gene in laryngeal carcinoma tissues, tissues adjacent to the tumor and normal laryngeal tissues. Hypermethylation of MGMT gene was detected in 16 samples of 46 (34.8%) laryngeal squamous cell carcinoma samples. However, the MGMT hypermethylation was not detected in all tissues adjacent to the tumors and normal tissues. No significant difference in MGMT gene hypermethylation was found in samples with different histological grades (chi2 = 3.130, P = 0.077) or in samples from patients with different TNM status (chi2 = 3.957, P = 0.138). No expression of MGMT mRNA was detected in all hypermethylated laryngeal carcinoma tissues. The expression of MGMT mRNA was detected in all unmethylated laryngeal carcinoma tissues, tissues adjacent to the tumors and normal tissues. It suggests that MGMT gene promoter hypermethylation is associated with MGMT gene transcription loss in laryngeal carcinoma tissues and possibly plays an important role in carcinogenesis of laryngeal tissues.
Carcinoma, Squamous Cell/*genetics ; CpG Islands/genetics ; DNA Methylation ; DNA Repair ; Laryngeal Neoplasms/*genetics ; O(6)-Methylguanine-DNA Methyltransferase/*genetics ; Polymerase Chain Reaction/methods ; Promoter Regions (Genetics)/*genetics

OBJECTIVE: To study the relationship between genetic polymorphisms of GSTM1 GSTT1 and the susceptibility of laryngeal and hypopharyngeal carcinomas (LHC). METHOD: The GSTM1 and GSTT1 genotypes were determined by multiplex PCR analysis in 76 LHC patients and 76 population controls. The association between the genotypes and LHC risk was measured by odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULT: The frequency of GSTM1 null genotype was 59.2% in the LHC patients and 42.1% in controls (OR=1.935, 95% CI=1.069-3.510), the difference was significant (P<0.01). The frequency of GSTT1 null genotype was 57.9% in the LHC patients and 51.3% in controls. The difference was not significant (P>0.05). In smokers, the risk of the LHC increased in subjects of GSTM1 null genotype (OR=5.545, 95% CI=2.158-13.528). CONCLUSION GSTM1 polymorphisms are associated with susceptibility to the LHC. It has the synergistic effects with smoking in the development of the LHC. GSTT1 genotypes might have no association with risk of the LHC in urban Linyi.
Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; Disease Susceptibility ; Female ; Gene Frequency ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Hypopharyngeal Neoplasms ; genetics ; Laryngeal Neoplasms ; genetics ; Male ; Polymorphism, Genetic

OBJECTIVE: To investigate the effect of sputum ubiquitin ligase (Cbl-b) gene known-down on the cytotoxicity of H9 T lymphocytes against human laryngeal squamous cancer Hep-2 cells and explore the underlying mechanism. METHODS: CD4 T lymphocytes isolated from 12 patients with laryngeal squamous carcinoma and 12 healthy individuals were examined for Cbl-b mRNA expressions using RT-PCR. H9 T lymphocytes cultured in 96-well plates were transfected with Cbl-b siRNA via liposomes followed by treatment with an anti-IL-2 monoclonal antibody, with H9 T lymphocytes transfected with a scrambled sequence as the negative control. The expressions of Cbl-b mRNA and protein in the cells were detected using real-time fluorescent quantitative PCR and Western blotting, respectively. The killing effect of the treated T lymphocytes against Hep-2 cells was assessed using the cell counting kit (CCK-8). The positive expression rates of CD69 and CD25 on the surface of H9 T lymphocytes were determined using flow cytometry, and the levels of interleukin-2 (IL-2) and interferon-gamma (INF-γ) in the culture supernatants of H9 T lymphocytes were detected with ELISA. RESULTS: The CD4 T lymphocytes from patients with laryngeal squamous carcinoma showed significantly increased Cbl-b mRNA level compared with those from healthy individuals ( < 0.05). Transfection of H9 T lymphocytes with Cbl-b siRNA significantly reduced the expression levels of Cbl-b mRNA and protein ( < 0.05), which were not significantly affected by subsequent treatment of the cells with the anti-IL-2 antibody (>0.05). At different target-effector ratios, the Cbl-b siRNA-transfected cells showed significantly higher Hep-2 cell killing rates and higher positivity rates of CD69 and CD25 expressions than the blank and negative control cells and the cells with both Cbl-b siRNA transfection and anti-IL-2 treatment ( < 0.05). Cbl-b silencing in H9 T lymphocytes resulted in significantly increased levels of IL-2 and INF-γ in the supernatant as compared with those in the blank and negative control groups ( < 0.05). CONCLUSIONS Cbl-b gene silencing effectively enhances the killing effect of H9 T lymphocytes against Hep-2 cells probably as the result of enhanced IL-2 secretion and T lymphocyte activation.
Carcinoma, Squamous Cell ; genetics ; therapy ; Gene Silencing ; Humans ; Laryngeal Neoplasms ; genetics ; therapy ; Lymphocyte Activation ; RNA, Small Interfering ; T-Lymphocytes