OBJECTIVEWith the objective of discovering novel putative chromosomal regions and special genes involved in the carcinogenesis, progression and metastasis of laryngeal squamous cell cancer (LSCC).

METHODSDNA copy profile of LSCC were obtained and analyzed by comparative genomic hybridization (CGH) and a computerized digital image analysis system. cDNA microarray of LSCC was performed and the profile was analyzed by Hierarchical clustering.

RESULTSCGH analysis showed average-12.9 gains and losses of chromosomes in LSCC. Relatively high frequencies of gains were found at 3q15-21 (14/18), 5p12-13 (11/18), 8q22-24 (6/18), 11q12-13 (8/18), 15q21-23 (7/18) and 18p11 (8/18), while those of losses at 1p13-21 (8/18), 3p21-23 (14/18), 5q21-22 (14/18), 9p12-pter (11/18) and 13q21-31 (8/18). Hierarchical clustering analysis showed that the differentially expressed genes were segregated into three groups. Three genes differentially expressed in process I (normal tissue to cancer) and process II (cancer to lymph node metastasis), and the Cy5/Cy3 ratios of twelve genes were either higher than 5.0 or lower than 0.2 in process I or process II. The fifteen special genes were first reported possibly to be the relationships with LSCC. In particular, 4 genes of them, which were cytochrome C oxidase Va, PPBP, EPHX2 and PON1, were first reported to correlate with tumorigenesis. SH3GL2, which was one of the 15 special genes, was located at one of the special chromosome regions, 9p12-pter.

CONCLUSIONThe important genes and special chromosomal aberrances might provide us a clue for further investigation of carcinogenesis, progression and metastasis in LSCC.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Chromosome Aberrations ; DNA, Neoplasm ; analysis ; Disease Progression ; Female ; Gene Expression ; Humans ; Karyotyping ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis

BACKGROUNDThe role of epigenetics in gene expression regulation and development significantly enhances our understanding of carcinogenesis. All the tumor related genes may be the target of epigenetical or genetic regulation. We selected some epigenetically regulated genes for cDNA array analysis and observed variability in the expression of the DICER1 gene in distinct stages of gastric cancer. The aim of this study was to assess the correlation between the expression of DICER1, an epigenetically regulated gene, and gastric cancer.

METHODSTo detect the expression of 506 tumor-associated genes, including DICER1, in the matched cancerous mucosa, pre-malignant lesion (adjacent mucosa), non-cancerous gastric mucosa and distant lymphocyte metastatic lesion in 3 cases of gastric cancers using cDNA array. DICER1 mRNA expression and DICER1 protein expression were further analyzed by Real-time PCR and Western blot in 32 cases of progressive gastric cancer. DICER1 protein expression was also detected in 33 early and 30 progressive gastric cancers by the immunohistochemistry (IHC) method.

RESULTSIn 3 cases of gastric cancer cDNA array showed dramatically decreased expression of DICER1 in pre-malignant lesion, cancerous mucosa and distant lymphocyte metastatic lesions compared with matched noncancerous gastric mucosa, pre-malignant lesion and cancerous mucosa. Real-time PCR results showed that the expression level of DICER1 mRNA in gastric cancer was significantly down-regulated compared to normal gastric tissue (P < 0.05). The IHC assay also showed that the expression of DICER1 was significantly decreased in progressive gastric cancer. Among the 63 cases of gastric cancers, 13/33 early (39.4%) and 19/30 (63.3%) progressive cancers showed negative expression of DICER1 (50.8%). The difference in expression of DICER1 between early and progressive gastric cancers was significant (P < 0.01). The result of Western blotting showed that DICER1 protein was down-regulated significantly in advanced gastric cancer (P < 0.05).

CONCLUSIONSDICER1 expression is decreased during the progression of gastric cancer, especially in progressive gastric cancers, which indicating DICER1 may play an important role in the development of cancer and the epigenetical regulation involved.

Blotting, Western ; DEAD-box RNA Helicases ; analysis ; genetics ; physiology ; Endoribonucleases ; analysis ; genetics ; physiology ; Epigenesis, Genetic ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Ribonuclease III ; Stomach Neoplasms ; chemistry ; etiology ; genetics

OBJECTIVETo explore mechanism of S100A8 in the oncogenesis and development of laryngeal cancer.

METHODSProteins interacting with S100A8 were isolated from laryngeal cancer cell lines Hep-2 by immunoprecipitation assay with anti-S100A8 antibody. The target bands were cut out and identified by maxtrix assisted laser desorption/ionization time of flight (MALDI-TOF). The peptide mass fingerprinting data of the proteins identified were analyzed based on the Mascot database. The NF-kappa B binding sites of the proteins were predicted by P-Match software. The binding ability of one of the proteins to S100A8 was confirmed by co-immunoprecipitation and immunocytochemistry methods.

RESULTSFour proteins interacting with S100A8 were obtained, which were hypothetical protein LOC80154, MHC class I HLA-B, similar to T-box 1 isoform C and sarcolemmal associated protein 1. The four genes were predicted to have NF-kappa B binding sites. MHC class I HLA-B, which is one of targets in NF-kappa B pathway, was first confirmed to have the binding ability to S100A8.

CONCLUSIONThe novel partners of S100A8 identified in the study might be involved in NF-kappa B pathway. The binding ability of MHC class I HLA-B to S100A8 implies that S100A8 might function as a new member with other proteins including HLA-B in NF-kappa B pathway. These findings provide a new clue to further study on the molecular mechanism of S100A8 in the genesis of laryngeal carcinomas.

Animals ; Binding Sites ; Calgranulin A ; genetics ; metabolism ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; HLA-B Antigens ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; NF-kappa B ; metabolism ; Signal Transduction

BACKGROUNDLaryngeal carcinoma is a common malignant tumor of the upper respiratory tract, and in 95% of cases the tumor is laryngeal squamous cell carcinoma (LSCC). The abnormity of SH3-domain GRB2-like 2 (SH3GL2) gene was found in LSCC. In order to clarify the relationship between SH3GL2 gene and LSCC, we evaluated the expression of the SH3GL2 gene in LSCC.

METHODReal-time PCR, immunohistochemistry and Western blotting were used to detect the mRNA and protein expression and find the various rules of SH3GL2 gene in LSCC.

RESULTSThe result of real-time PCR showed that the expression level of SH3GL2 mRNA in LSCC tissue was apparently down-regulated; immunohistochemical analysis showed that SH3GL2 protein was mainly located in cytoplasm, the rate of positive cells and SH3GL2 protein expression level were fluctuated with the pathological classification of LSCC; the result of Western blotting showed that SH3GL2 protein was down-regulated significantly in LSCC samples, especially in metastatic lymph nodes.

CONCLUSIONSThese results suggest that SH3GL2 is a LSCC related gene and its expression level is fluctuated with the pathological classification which indicate that SH3GL2 participates in the development and progression of LSCC. And it may be considered as a novel tumor marker to find both a new anti-oncogene and relative factors of invasion and metastasis of laryngeal carcinoma.

Adaptor Proteins, Signal Transducing ; analysis ; genetics ; Blotting, Western ; Carcinoma, Squamous Cell ; chemistry ; genetics ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; chemistry ; genetics ; Polymerase Chain Reaction ; src Homology Domains

OBJECTIVETo study the use of neural network in determining the risk factors of diseases.

METHODSWith back-propagation neural network (BP network) as fitting model based upon data gathered from an epidemiological survey on diabetes mellitus and under the network structure of 22-6-1, the mean impact value (MIV) for each input variables and sequencing the factors according to their absolute MIVs were calculated. The results from BP network with multiple logistic regression analysis and log-linear model for united actions between factors were compared with optimizing Levenberg-Marquardt algorithm.

RESULTSBy BP network analysis, the sequence of importance for the risk factors of diabetes mellitus became: faster pulse, diabetes mellitus family history, living longer in the investigated area, with medical record of nephropathy, having higher ratio for waist-to-hip, being male, with medical records of diseases as hyperlipoproteinmia, coronary heart disease, hypertension, high diastolic pressure, higher income, do no drink alcohol, age, higher systolic pressure, less educated, body mass index, with medical records of other diseases, physical exercise related to jobs smoking, occupation, with medical record for cerebrovascular disease, with medical record for liver disease etc. However, only 7 factors were statistically significant in multiple logistic regression analysis. The sequence of their importance appeared as: pulse, diabetes mellitus family history, the medical record of nephropathy, waist-to-hip ratio, the medical record of hypertension, work-place related exercise and age. The sequences of importance were almost the same between the two while the difference could partly be explained by the interaction among risk factors through log-linear model.

CONCLUSIONNeural network could be used to analyze the risk factors of diseases and could assimilate more complicated relationships (main effects and interactions) between inputs and outputs, better than using the traditional methods.

Adult ; China ; epidemiology ; Diabetes Mellitus ; epidemiology ; etiology ; Family Health ; Female ; Humans ; Hyperlipidemias ; complications ; Logistic Models ; Male ; Neural Networks (Computer) ; Obesity ; complications ; Pulse ; Risk Factors

Objective To observe the levels of serum interleukin-1β(1L-1β)and interleukin-1 receptor antagonist(IL-1Ra),the IL-1Ra/IL-1β ratio and the relationship between the levels of thern and the restenosis in patients with intra-and extra-cranial arteriostenosis after stent-assisted angioplasty.Methods Thirty-one patients with cerebral artery stenosis,admitted to our hospital from April 2003 to March 2006,were treated with stent-assistant angioplasty and followed up for 6-12 months.The relationship was analyzed between the restenosis of cerebral artery and both the levels of serum IL-1β,IL-1Ra and the IL-1 Ra/IL-1β ratio before and 1 h,1,3 and 5 d after stent-assisted angioplasty.Results The condition of 31 stents in 31 blood vessels of cerebral arteries was observed and followed up 6 to 12 months after the operation.Restenosis was noted in 6(19.3%)with 3 restenosis superior to 50% and 3 restenosis from 10%-30%.No obvious difference of the levels of serum IL-1β and IL-1Ra,the IL-1Ra/IL-1β ratio between restenosis group and non-restenosis group before the operation was found (P>0.05);IL-11β and IL-1Ra levels were positively correlated before the operation.The levels of serum IL-1β and IL-1Ra 1h,1,3 and 5 d after the operation were superior to those before the operation,respectively(P<0.05).The levels of serum IL-1β and IL-1Ra in the restenosis group showed no significant difference to those in the non-restenosis group 6 to 12 months alter the operation(P>0.05);however,the IL-1Ra/IL-1β ratio in the restenosis group was significantly lower than that in the non-restenosis group (P<0.05);positive correlation between the levels of IL-1β and IL-1Ra was observed.Conclusion The levels of serum IL-1β and IL-1Ra were associated with the inflammatory process after the operation and the immune maladjustment of IL-1β and IL-1Ra might be correlative to the restenosis,indicating that the IL-1Ra/IL-1β ratio might be an available index for monitoring the restenosis.

Objective To discuss mechanism and control measures of stent fracture and restenosis after percutaneous transluminal angioplasty and stenting (PTAS) for symptomatic ostial vertebral/subclavian artery stenosis. Methods A retrospective analysis was performed on 3 patients with stent fracture after receiving PTAS for symptomatic ostial vertebral/subclavian artery stenosis.Simple radiographic, ultrasonographic and clinical follow-up examinations were estimated. Related articles on coronary stent fracture were gone over, consulting in the types, cumulative incidence and occurrence time of adverse events, risk factors and preventive measures. Results Stent fractures of 3 patients with symptomatic ostial vertebral /subclavian artery stenosis were associated with in-stent restenosis and occlusion. Two of the 3 patients treated with the balloon angioplasty and after balloon dilatation, and the patients exhibited relief of symptoms. One patient was only managed for vascular disease risk factors, and no developing recurrent symptoms were noted during the follow-up period.Conclusions Stent fracture might appear in patients performed PTAS for symptomatic ostial vertebral /subclavian artery stenosis, and regular check is needed. Individual treatment was emphasized in case of serious symptoms appeared.

OBJECTIVETo investigate the role of STK15 gene amplification and overexpression to genesis and development of laryngeal squamous cell carcinoma (LSCC).

METHODSSTK15 gene amplification in 40 cases carcinoma tissues and normal tissues as control was detected by differential PCR approach. STK15 mRNA and protein levels were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry method.

RESULTSIn 40 LSCC cases, STK15 gene amplification was found in 14 tumor tissues(35%), mRNA overexpression in 27 tumor tissues(67.5%), and protein upregulated in 29 tumor tissues(72.5%). Statistics analysis showed that STK15 gene amplification and mRNA overexpression were obviously associated to differentiation degree of LSCC, and protein overexpression was closely associated with both differentiation degree and pathological grades of LSCC.

CONCLUSIONThis research results suggest that STK15 gene amplification contributes to its mRNA and protein overexpression through affecting the exact replication of centrosome and separation of chromosomes. STK15 gene thus plays a role in LSCC oncogenesis and malignant progression.

Aurora Kinase A ; Aurora Kinases ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

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.

Cell Line, Tumor ; Chromosome Aberrations ; Humans ; In Situ Hybridization, Fluorescence ; Laryngeal Neoplasms ; genetics ; pathology ; Tumor Cells, Cultured

OBJECTIVETo explore the relationship between p53 gene mutations and STK15 abnormal expression in the development of human laryngeal squamous-cell carcinoma (LSCC).

METHODSLSCC tissues and matched normal tissues were taken during operation from 55 patients without previous chemotherapy or radiotherapy. Following polymerase chain reaction amplification direct sequencing single strand conformational polymorphism (PCR-SSCP) combined with silver staining were used to detect mutations of p53 gene in exons 7 and 8 (p53E7 and p53E8) using genomic DNA from 110 specimens including 55 LSCC tissues and 55 matched normal tissues. STK15 expression were evaluated by RT-PCR with beta-actin as internal control.

RESULTSThe mutation rate of p53E7 was 30.9% (compared to normal tissues, chi(2) = 8.66, P < 0.01). There was no mutation in p53E8. In 38 of the 55 cases (69.1%), the STK15 mRNA expression level was higher than that of the paired normal tissue. The STK15 to beta-actin ratio of average density value was 1.22 +/- 0.49 in the cancer tissue, and 0.99 +/- 0.54 in the normal tissues (t = 4.539, P < 0.01). In 14 of the 17 cases (82.4%) with p53E7 mutations, the STK15 expression was higher than that of normal tissue. In the 38 cases with STK15 over-expression, p53E7 mutation was found in 14 cases (36.8%). The rate of concurrence of p53E gene mutations and STK15 over-expression (25.5%) was higher than that of only p53E gene mutations (chi(2) = 26.025, P < 0.01).

CONCLUSIONThere is significant association between p53 gene mutation and STK15 over-expression in laryngeal squamous-cell carcinoma.

Actins ; metabolism ; Aurora Kinase A ; Aurora Kinases ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Exons ; Frameshift Mutation ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; genetics ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; Mutation, Missense ; Protein-Serine-Threonine Kinases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics