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PD.127 Expression of RAD21 cleaved products in oral epithelium
Podium Sessions
104
study aimed to test the hypothesis that indMduals with the 2G/2G genotype of MMP-1 promoter polymorphism may be at an increased risk for head and neck squamous cell carcinoma (HNSCC); and the disease aggressiveness should be greatly elevated m HNSCC patients with 2G genotype. Materials and Methods: The 250 patients newly diagnosed with HNSCC and 250 cancer-free controls that consented to participate in a hospital-based, case-control study were interviewed with a structured questionnaire and provided blood. DNA was extracted for genotypmg using a PCR-RFLP assay. The levels of MMP-1 expression m HNSCC specimens were evaluated by the qnantitative RT-PCR and the correlation between different genotype and MMP-1 expression was determined. Odds ratio were calculated using mulfivariate logistic regression. In addition, the underlying mechanism responsible for MMP-1 regulation m HNSCC was exasmned by the coculture system. Results: The controls were frequency-niatched to the cases by age (+ 5 years), sex and smokmg/drHkk_mg status. SubJects carrying the homozygous 2G/2G MMP-1 genotype had a fold hiooher risk of head and neck cancer compared with subjects with other genotypes (odds ratio = 1.82, 95% confidence interval: 1.30- 2.45), controlling for major confounders. A correlation between promoter polyniorphasm and MMP-1 expression was found, and that this SNP was correlated with the clinical stage of HNSCC. Finally the tumor-fibroblast coculture assay revealed that fibroblasts with 2G/2G genotype enhanced the MMP-1 expression to the highest level. Conclusion: We demonstrated for the first t ~ i e the distribution and tire clinical significance of MMP-1 genotypes m HNSCC patients. These fin4uigs suggested that SNP of MMP-1 promoter might influence the ability m HNSCC invasion via transcriptional activity of this gene.
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Expression of RAD21 cleaved products in oral epithelium
G. Yanianioto, Y. Nagost~, A. Fukuda, T. Aida, T. Irie, T. Tachlkawa. Departmoa of Oral Pathology and Dmgnoszs,
57towa Unmerstty Sd~ool of Domst~y. Japan Introduetion: RAD21 as involved an the repair of doublestrand breaks an DNA and as essential for ~mtotic growth. Recently, it has been shown that the degradation product formed by the C-terminal cleavage of RAD21 by caspases 3 and 7 becomes a factor which reduces apoptosis. In addition, although at is known that there are n~aay cleavage products, about those functions, ~t is not clear. In this study, tire relevance of RAD21 cleaved products expression to the defferentiation and carcinogenesis of oral epithelium was clarified using Laser Mlcrodlssectlon and Western Blot. Materials and Methods: We used the SAS cell, oral squamous cell carcinoma cell line. The cells were treated with retmolc acid, ALLN and Z-VAD-FMK at the desired final concentration and duration. And the protein was extracted from each culture cells. In addition, the protein was extracted by Laser Microdlssection from the frozen sections of normal eplthehum, leukoplakia, epithelial dysplasla, squamous cell carcinoma of oral region. Each samples were analyzed by Western Blot using Anti-Rid 21 rabbit polyclonal antibody. Results: We detected Rad 21 cleaved products from the protein extracted by Laser Microdlssection from the frozen sections. One of products was detected in the cell treated by retinoic acid mid leukoplakla, and some products vamshed m normal epithehum. Conclusion: Laser Mlerodissection and Western Blot are a very useful method for analyzing cleaved products in a tissue
section. Our findings suggest that the RAD21 cleaved products play an important role in the defferentiafion and carcinogenesis of oral epithelium.
PD.128 Gene
expression profiles of oral squamous cell carcinoma depending on lymph node status
T. Fillies 1, H. Bfirger 2, B. Brandt s, D. Kemming s, R. Werkmeister 4, U. Joos 1, J. Klemheinz 1. Department
of Cranio-Maxillofacial Sm'ger): Unmers~ty qf MiinsteT; Germany Introduction: Cancer represents the phenotypic end point of multiple genetic lesions that endow cells with a full range of biological properties required for tumorgenesis. Molecular changes of tumor cells play an important role m tumor development and progression. The objective of this study was to identify the change of expression profiles of oral sqnamous cell carcinoma (OSCC) for lymph node positive compared to lymph node negative status. Materials and Methods: We included ei~lat patient with T2 OSCC of which were four lymph node posilave and four lymph node negative. We analyzed the differential gene expression profiles of OSCC comparing lymph node positive with lymph node negative group applying statistical procedures such as t-test, F-test and fold change for group comparison. Gene expression profiles were generated using Applied Blosystem Genome Survey Microarray, comprising 31,000 genes. Results: With this method we identified 15 genes that were expressed differently with statistically significance and that showed multiple fold change between squamous cell carcinoma lymph node negative and lymph node positive OSCC Conclusion: Dlfferent,al expression pattern of lymph node positive and lymph no de negative oral squasnous cell carcmonm may have an unpact on the potential ability for lymph node metastasis m OSCC. Further analysis may provide a better understan4mg of the biology of oral squamous cell carcinoma metastasis.
•]
DCC gene inactivation in head and neck squamous carcinoma
A.L. Carvatho, W.-W. Jlang, S. Begum, M. Zhao, M. Brait, C. Jerommo, R. Henrlque, S. Chang, W. Koch, W.H. Westra, J.A. Cal~ano. Johns Hopl,~ns Unmerstt); Department of
Otola~yngology-Head cmd Neck Ntrgery. USA Introduction: DCC is a candidate tumor suppressor gene located at chromosome 18@1 that has been lmphcated m colorectal tumorigenesis, but its role m head and neck squamous cell carcinoma (HNSC) is not well described. We evaluated DCC promoter tlypermethylation, loss of heterozygosity (LOH), and expression m head and neck squan~ous carcinoma and their correlation. Materials and Methods: Tumor and lymphocyte DNA were obtained from 56 patients. For hypermethylation analysis, tumor DNA underwent blsulfite treatment and was analyzed m a quantitative fashion using real-rune quantitative MSP (QMSP). For LOH analysis, allehc imbalance was investigated comparing tumor and lymphocyte DNA regarding loci inside chromosome 18, including loci in the DCC region. DCC expression was also evaluated by immunohistochemisty (IHC). Results: Regarding the Q-MSP analysis, 42 cases (75.0%) showed DCC promoter region hypermethylatlon. LOH was present m 13 cases (23.2%), 6 of thenl being m DCC region. IHC showed focal DCC expression m 13 cases (23.2%), diffuse expression in 14 cases (25.0%), and no expression in 29 cases
104
study aimed to test the hypothesis that indMduals with the 2G/2G genotype of MMP-1 promoter polymorphism may be at an increased risk for head and neck squamous cell carcinoma (HNSCC); and the disease aggressiveness should be greatly elevated m HNSCC patients with 2G genotype. Materials and Methods: The 250 patients newly diagnosed with HNSCC and 250 cancer-free controls that consented to participate in a hospital-based, case-control study were interviewed with a structured questionnaire and provided blood. DNA was extracted for genotypmg using a PCR-RFLP assay. The levels of MMP-1 expression m HNSCC specimens were evaluated by the qnantitative RT-PCR and the correlation between different genotype and MMP-1 expression was determined. Odds ratio were calculated using mulfivariate logistic regression. In addition, the underlying mechanism responsible for MMP-1 regulation m HNSCC was exasmned by the coculture system. Results: The controls were frequency-niatched to the cases by age (+ 5 years), sex and smokmg/drHkk_mg status. SubJects carrying the homozygous 2G/2G MMP-1 genotype had a fold hiooher risk of head and neck cancer compared with subjects with other genotypes (odds ratio = 1.82, 95% confidence interval: 1.30- 2.45), controlling for major confounders. A correlation between promoter polyniorphasm and MMP-1 expression was found, and that this SNP was correlated with the clinical stage of HNSCC. Finally the tumor-fibroblast coculture assay revealed that fibroblasts with 2G/2G genotype enhanced the MMP-1 expression to the highest level. Conclusion: We demonstrated for the first t ~ i e the distribution and tire clinical significance of MMP-1 genotypes m HNSCC patients. These fin4uigs suggested that SNP of MMP-1 promoter might influence the ability m HNSCC invasion via transcriptional activity of this gene.
•
Expression of RAD21 cleaved products in oral epithelium
G. Yanianioto, Y. Nagost~, A. Fukuda, T. Aida, T. Irie, T. Tachlkawa. Departmoa of Oral Pathology and Dmgnoszs,
57towa Unmerstty Sd~ool of Domst~y. Japan Introduetion: RAD21 as involved an the repair of doublestrand breaks an DNA and as essential for ~mtotic growth. Recently, it has been shown that the degradation product formed by the C-terminal cleavage of RAD21 by caspases 3 and 7 becomes a factor which reduces apoptosis. In addition, although at is known that there are n~aay cleavage products, about those functions, ~t is not clear. In this study, tire relevance of RAD21 cleaved products expression to the defferentiation and carcinogenesis of oral epithelium was clarified using Laser Mlcrodlssectlon and Western Blot. Materials and Methods: We used the SAS cell, oral squamous cell carcinoma cell line. The cells were treated with retmolc acid, ALLN and Z-VAD-FMK at the desired final concentration and duration. And the protein was extracted from each culture cells. In addition, the protein was extracted by Laser Microdlssection from the frozen sections of normal eplthehum, leukoplakia, epithelial dysplasla, squamous cell carcinoma of oral region. Each samples were analyzed by Western Blot using Anti-Rid 21 rabbit polyclonal antibody. Results: We detected Rad 21 cleaved products from the protein extracted by Laser Microdlssection from the frozen sections. One of products was detected in the cell treated by retinoic acid mid leukoplakla, and some products vamshed m normal epithehum. Conclusion: Laser Mlerodissection and Western Blot are a very useful method for analyzing cleaved products in a tissue
section. Our findings suggest that the RAD21 cleaved products play an important role in the defferentiafion and carcinogenesis of oral epithelium.
PD.128 Gene
expression profiles of oral squamous cell carcinoma depending on lymph node status
T. Fillies 1, H. Bfirger 2, B. Brandt s, D. Kemming s, R. Werkmeister 4, U. Joos 1, J. Klemheinz 1. Department
of Cranio-Maxillofacial Sm'ger): Unmers~ty qf MiinsteT; Germany Introduction: Cancer represents the phenotypic end point of multiple genetic lesions that endow cells with a full range of biological properties required for tumorgenesis. Molecular changes of tumor cells play an important role m tumor development and progression. The objective of this study was to identify the change of expression profiles of oral sqnamous cell carcinoma (OSCC) for lymph node positive compared to lymph node negative status. Materials and Methods: We included ei~lat patient with T2 OSCC of which were four lymph node posilave and four lymph node negative. We analyzed the differential gene expression profiles of OSCC comparing lymph node positive with lymph node negative group applying statistical procedures such as t-test, F-test and fold change for group comparison. Gene expression profiles were generated using Applied Blosystem Genome Survey Microarray, comprising 31,000 genes. Results: With this method we identified 15 genes that were expressed differently with statistically significance and that showed multiple fold change between squamous cell carcinoma lymph node negative and lymph node positive OSCC Conclusion: Dlfferent,al expression pattern of lymph node positive and lymph no de negative oral squasnous cell carcmonm may have an unpact on the potential ability for lymph node metastasis m OSCC. Further analysis may provide a better understan4mg of the biology of oral squamous cell carcinoma metastasis.
•]
DCC gene inactivation in head and neck squamous carcinoma
A.L. Carvatho, W.-W. Jlang, S. Begum, M. Zhao, M. Brait, C. Jerommo, R. Henrlque, S. Chang, W. Koch, W.H. Westra, J.A. Cal~ano. Johns Hopl,~ns Unmerstt); Department of
Otola~yngology-Head cmd Neck Ntrgery. USA Introduction: DCC is a candidate tumor suppressor gene located at chromosome 18@1 that has been lmphcated m colorectal tumorigenesis, but its role m head and neck squamous cell carcinoma (HNSC) is not well described. We evaluated DCC promoter tlypermethylation, loss of heterozygosity (LOH), and expression m head and neck squan~ous carcinoma and their correlation. Materials and Methods: Tumor and lymphocyte DNA were obtained from 56 patients. For hypermethylation analysis, tumor DNA underwent blsulfite treatment and was analyzed m a quantitative fashion using real-rune quantitative MSP (QMSP). For LOH analysis, allehc imbalance was investigated comparing tumor and lymphocyte DNA regarding loci inside chromosome 18, including loci in the DCC region. DCC expression was also evaluated by immunohistochemisty (IHC). Results: Regarding the Q-MSP analysis, 42 cases (75.0%) showed DCC promoter region hypermethylatlon. LOH was present m 13 cases (23.2%), 6 of thenl being m DCC region. IHC showed focal DCC expression m 13 cases (23.2%), diffuse expression in 14 cases (25.0%), and no expression in 29 cases