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1.
Int J Oncol ; 47(4): 1249-56, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26323893

ABSTRACT

Increasing evidence indicates that cancer stem cells have essential roles in tumor initiation, progression, metastasis and resistance to chemo-radiation. Recent research has pointed out biological importance of microRNAs in cancer stem cell dysregulation. Total number of mature microRNAs in human genome increased to more than 2,500 with the recent up-date of the database. However, currently no information is available regarding microRNA expression profiles of cancer stem cells in head and neck squamous cell carcinoma (HNSCC). Increased ALDH1 activity has been demonstrated as a reliable marker for isolation of cancer stem cells. Therefore, we evaluated the microRNA expression profile of ALDH1-high subpopulations in the HNSCC cell lines UTSCC-9 and UTSCC-90. Initially, we examined cancer stem cell properties of ALDH1-high subpopulations in both cell lines. We analyzed expression of stemness markers, sphere formation capacity and xenograft transplantation into NOD/SCID mice. Our findings validated that ALDH1-high subpopulations showed significantly increased tumor-initiating ability. Furthermore, we investigated the microRNA expression profile of HNSCC stem cells using microRNA array and confirmed the results by quantitative real-time PCR. We found that expressions of miR­424, let-7a, miR­6836, miR­6873 and miR­7152 were downregulated, whereas miR­147b was upregulated with statistical significance in the ALDH1-high subpopulation. In conclusion, we identified a subset of microRNAs that were differentially expressed in ALDH1-high subpopulation, providing new microRNA targets to study dysregulation of HNSCC-initiating cells and develop therapeutic strategies aimed at eradicating the tumorigenic stem cells in HNSCC.


Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/pathology , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/pathology , MicroRNAs/analysis , MicroRNAs/genetics , Neoplastic Stem Cells/pathology , Animals , Cell Line, Tumor , Cell Separation , Female , Gene Expression Regulation, Neoplastic , Heterografts , Humans , Mice , Mice, Inbred NOD , Mice, SCID , Oligonucleotide Array Sequence Analysis , Real-Time Polymerase Chain Reaction , Squamous Cell Carcinoma of Head and Neck , Transcriptome
2.
Int J Mol Med ; 33(4): 879-86, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24424572

ABSTRACT

Epithelial-mesenchymal-transition (EMT) is a critical step in tumor invasion and metastasis, while its fate is mainly defined by the balanced expression between the miR-200 family and ZEB transcription factors. In this study, we observed a reciprocal correlation between miR-200c/mir-141 and ZEB1, as well as between ZEB2 and E-cadherin expression in a panel of 13 head and neck squamous cell carcinoma (HNSCC) cell lines. We also confirmed that the enforced expression of miR-200c and miR-141 significantly reduced the migration capacity of HNSCC cells. Accordingly, the enforced expression of miR-200c and mir-141 resulted in a significant upregulation in E-cadherin expression, contrary to the significant downregulation in ZEB1 expression in 3 cell lines (UTSCC-24A, UTSCC-24B and UTSCC-6A cells). Another pair of cell lines, UTSCC-60A and UTSCC­ 60B failed to show a significant change in the expression of E-cadherin or ZEB1/ZEB2 during the enforced expression of miR-200c/miR-141. To address the issue, we focused on the hypermethylation status of the ZEB1/2 promoters, which have both been shown to include wide CpG islands. We observed a marked upregulation in both ZEB1 and ZEB2 mRNA expression following treatment with a demethylating agent in both pairs of UTSCC cell lines. In conclusion, our findings confirm the existence of a reciprocal correlation between the mir-200 family and the ZEB family, and demonstrate the role of the miR-200 family in EMT, as well as in the migration and invasion ability of HNSCC cells. Furthermore, our data suggest that the promoter hypermethylation of ZEB1 and ZEB2 may play an essential role and may overshadow the effects of the miR-200 family in the regulation of EMT during carcinogenesis.


Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/pathology , Cell Movement/genetics , Epithelial-Mesenchymal Transition/genetics , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/pathology , MicroRNAs/metabolism , Cadherins/metabolism , Cell Line, Tumor , Cell Proliferation , DNA Methylation/genetics , Gene Expression Regulation, Neoplastic , Homeodomain Proteins/metabolism , Humans , MicroRNAs/genetics , Neoplasm Invasiveness , RNA, Messenger/genetics , RNA, Messenger/metabolism , Repressor Proteins/metabolism , Squamous Cell Carcinoma of Head and Neck , Transcription Factors/metabolism , Zinc Finger E-box Binding Homeobox 2 , Zinc Finger E-box-Binding Homeobox 1
3.
Oncol Lett ; 3(2): 273-280, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22740894

ABSTRACT

Head and neck squamous cell carcinoma (HNSCC) is one of the most frequently occurring types of cancer worldwide. We focused on the fact that the aberrant function of Wnt/ß-catenin signaling is a frequent event in malignancies. Dickkopf (Dkk)-3 is a major negative regulator of Wnt/ß-catenin signaling, which is a known tumor suppressor and is down-regulated in various types of cancer. However, the expression profile of the Dkk-3 protein in HNSCC has not yet been reported. The present study was conducted to investigate Dkk-3 protein expression in 90 cases of HNSCC tissue samples and HNSCC-derived cell lines. In contrast to findings available on other types of cancer, the Western blot analysis revealed that HNSCC cell lines expressed the Dkk-3 protein. In immunohistochemistry, 76 cases (84.4%) out of 90 tissue samples were Dkk-3-positive, whereas only 14 cases (15.6%) were negative. Notably, survival analysis showed that the Dkk-3 (-) group exhibited significantly longer disease-free survival (p=0.038), metastasis-free survival (p=0.013) and longer overall survival (p=0.155). The results showed that the Dkk-3 protein was dominantly expressed and may be involved in carcinogenesis and metastasis in HNSCC. Moreover, the findings suggest that the function of Dkk-3 differs depending on the tissue of origin, and that it may exert an oncogenic function in HNSCC.

5.
Oncol Res ; 18(10): 509-18, 2010.
Article in English | MEDLINE | ID: mdl-20681410

ABSTRACT

Ameloblastoma is the most frequently encountered odontogenic tumor, characterized by a locally invasive behavior, frequent recurrences, and, although rare, metastatic capacity. Loss or inactivation of tumor suppressor genes (TSGs) allows cells to acquire neoplastic growth. The ING family proteins are tumor suppressors that physically and functionally interact with p53 to perform important roles in apoptosis, DNA repair, cell cycle regulation, and senescence. TP53 genetic alterations were reported to infrequently occur in ameloblastoma. Considering that other TSGs related to TP53 could be altered in this tumor, we focused our study on the ING family genes. We analyzed the loss of heterozygosity (LOH) status of the ING family (ING1-ING5) chromosomal loci in a group of ameloblastomas by microsatellite analysis, and correlated the ING LOH status with clinicopathological characteristics. By using specific microsatellite markers, high frequency of LOH was found at the loci of each ING gene family member (33.3-72.2%). A significant relationship was shown between LOH of D2S 140 (ING5 locus) and solid tumor type (p = 0.02). LOH of ING3MS (ING3 locus) was also high in solid type tumors, showing a near significant association. In addition, a notable tendency toward higher LOH for half of the markers was observed in recurrent cases. LOH of ING family genes appears as a common genetic alteration in solid ameloblastoma. The current study provides interesting novel information regarding the potential prognostic significance of the allelic loss of the ING gene family loci in ameloblastoma tumorigenesis.


Subject(s)
Ameloblastoma/genetics , Cell Cycle Proteins/genetics , Homeodomain Proteins/genetics , Intracellular Signaling Peptides and Proteins/genetics , Jaw Neoplasms/genetics , Loss of Heterozygosity , Nuclear Proteins/genetics , Trans-Activators/genetics , Tumor Suppressor Proteins/genetics , Adolescent , Adult , Aged , Ameloblastoma/pathology , Child , Female , Genes, Tumor Suppressor , Humans , Inhibitor of Growth Protein 1 , Male , Middle Aged
6.
Int J Cancer ; 127(9): 2088-94, 2010 Nov 01.
Article in English | MEDLINE | ID: mdl-20131318

ABSTRACT

Our previous study showed high frequency of allelic loss at chromosome 2q37 region in oral cancer. This location contains several candidate tumor suppressor genes such as PPP1R7, ILKAP, DTYMK and ING5. We previously showed 3 members of inhibitor of growth (ING) family, ING1, ING3 and ING4 as tumor suppressor gene in head and neck cancer. As ING5 shows high homology with other members of ING genes including highly conserved carboxy-terminal plant homeodomain and nuclear localization signal, we first picked up ING5 and examined it as a possible tumor suppressor in oral cancer. For this aim, mutation and mRNA expression status of ING5 in paired normal and oral squamous cell carcinoma samples were examined by reverse transcription polymerase chain reaction (RT-PCR) and sequencing. Three missense mutations located within leucine zipper like (LZL) finger and novel conserved region (NCR) domains in ING5 protein were detected, probably abrogating its normal function. We also found 5 different alternative splicing variants of ING5. Then, we examined mRNA level of ING5 by quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) analysis, which demonstrated decreased expression of ING5 mRNA in 61% of the primary tumors as compared to the matched normal samples. In conclusion, tumor-specific mutation and downregulation of ING5 mRNA suggested it as a tumor suppressor gene in oral squamous cell carcinoma.


Subject(s)
Carcinoma, Squamous Cell/genetics , Mouth Neoplasms/genetics , Transcription Factors/genetics , Tumor Suppressor Proteins/genetics , Down-Regulation , Humans , Mutation , RNA, Messenger/metabolism
7.
Cancer Invest ; 28(5): 495-500, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20014947

ABSTRACT

We analyzed mutation and expression status of human epidermal growth factor receptor 2 (Her2) in head and neck squamous cell carcinoma (HNSCC) using single strand conformation polymorphism (SSCP) mutation analysis and immunohistochemistry (IHC). Mutations were absent in all 85 cases. Out of 57 cases available for IHC, Her2 protein expression was negative (0) in 40 tumors (70%). Seventeen tumors (29.8%) expressed Her2, among these 13 tumors (22.8%) showed a weak (+1) expression and 4 (7%) showed a moderate expression (+2), none showed a strong (+3) expression. There was not a significant association between expression and any of the patients' clinical variables or prognosis. Our results suggest that Her2 may not be useful as a molecular target in HNSCC.


Subject(s)
Carcinoma, Squamous Cell/genetics , ErbB Receptors/genetics , Head and Neck Neoplasms/genetics , Carcinoma, Squamous Cell/mortality , DNA Mutational Analysis , Female , Head and Neck Neoplasms/mortality , Humans , Male , Middle Aged , Mutation
8.
Cancer Invest ; 28(1): 103-10, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19995224

ABSTRACT

Head and neck squamous cell carcinoma (HNSCC) is a frequently occurring cancer worldwide. Dickkopf (Dkk)-1 gene is suggested to function as tumor suppressor gene (TSG) in several kinds of malignancies. In this study, we performed loss of heterozygosity (LOH) analysis of Dkk-1 and examined the correlation between LOH status and clinicopathological parameters for the first time. A pretty high LOH ratio (50%) was detected. Interestingly, in the cases with Dkk-1 retention group showed less distant metastasis and a tendency of longer disease free survival. These results indicate that Dkk-1 can play a role in HNSCC carcinogenesis and it may also be related to distant metastasis.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 10 , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/genetics , Intercellular Signaling Peptides and Proteins/genetics , Loss of Heterozygosity , Aged , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/secondary , Carcinoma, Squamous Cell/therapy , Disease-Free Survival , Female , Gene Frequency , Head and Neck Neoplasms/mortality , Head and Neck Neoplasms/secondary , Head and Neck Neoplasms/therapy , Humans , Kaplan-Meier Estimate , Male , Microsatellite Repeats , Middle Aged , Time Factors , Treatment Outcome
9.
Auris Nasus Larynx ; 37(2): 137-44, 2010 Apr.
Article in English | MEDLINE | ID: mdl-19505782

ABSTRACT

OBJECTIVES: The aim of this study is to examine the internalization of nontypeable Haemophilus influenzae (NTHi) into human epithelial cells. METHODS: Bactericidal assay was applied to examine the effects of antibiotics against cell-adherent NTHi using HEp-2 cells. A trans-well chamber assay was applied to examine the internalization and penetration of NTHi using Detroit562 cells. RESULTS: The adherence of NTHi to HEp-2 cells was noted after 2h of incubation. Azithromycin had a strong bactericidal effect against both cell-associated and non-adherent NTHi, while ceftriaxone did not show bactericidal effects on NTHi adhered to the HEp-2 cells. Three (60.0%) out of five NTHi isolates from the nasopharynx of children with intractable acute otitis media (AOM) internalized into and subsequently penetrated through the epithelial cells at various degrees. Azithromycin had a strong bactericidal effect against the cell-internalized NTHi, while ceftriaxone was bactericidal only against extracellular NTHi. CONCLUSION: The potential of NTHi as the intracellular pathogen may contribute to the persistent existence of this pathogen that result in the prolonged and intractable clinical course of AOM. Azithromycin may be a therapeutically significant antibiotic for patients with prolonged respiratory tract infections due to NTHi.


Subject(s)
Anti-Bacterial Agents/pharmacology , Azithromycin/pharmacology , Bacterial Adhesion/drug effects , Bacterial Physiological Phenomena/drug effects , Ceftriaxone/pharmacology , Epithelial Cells/microbiology , Gentamicins/pharmacology , Haemophilus Infections/microbiology , Haemophilus influenzae/classification , Haemophilus influenzae/isolation & purification , Otitis Media with Effusion/microbiology , Acute Disease , Cell Line , Child , Dose-Response Relationship, Drug , Epithelial Cells/drug effects , Haemophilus Infections/drug therapy , Haemophilus influenzae/drug effects , Humans , In Vitro Techniques , Microbial Sensitivity Tests , Microbial Viability/drug effects , Otitis Media with Effusion/drug therapy
10.
Cancer Invest ; 27(6): 661-8, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19488910

ABSTRACT

We have analyzed allelic loss of the short arm of chromosome 9 in 39 head and neck cancers using 13 polymorphic markers and found two deletions of hot spots at 9p21 and 9p24. Loss of heterozygosity was detected at least at one locus in 28 of 39 cases (71.8%). P16, a well-known tumor suppressor gene, is considered to be a target for deletion of the 9p21 region. However, novel frequent chromosomal deletion and a candidate tumor suppressor gene, BRM at the 9p24 region, were detected. Moreover, comparison of clinicopathological variables demonstrated that loss of heterozygosity at the BRM locus was associated with a worse prognosis.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 9 , Gene Expression Regulation, Neoplastic , Genes, Tumor Suppressor , Head and Neck Neoplasms/genetics , Loss of Heterozygosity , Transcription Factors/genetics , Aged , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/therapy , Disease-Free Survival , Female , Genotype , Head and Neck Neoplasms/mortality , Head and Neck Neoplasms/pathology , Head and Neck Neoplasms/therapy , Humans , Kaplan-Meier Estimate , Male , Microsatellite Repeats , Middle Aged , Phenotype , Treatment Outcome
11.
Cancer Sci ; 100(5): 873-80, 2009 May.
Article in English | MEDLINE | ID: mdl-19445022

ABSTRACT

Previous gene expression profiles revealed the T-lymphocyte maturation-associated protein (MAL) gene as being frequently downregulated in head and neck cancer. To define the relationship between the MAL gene and the metastatic process, we evaluated the expression status of the gene in matched primary and metastatic tumors of head and neck cancer by semiquantitative reverse transcription-polymerase chain reaction. Furthermore, we aimed to identify potential genetic and epigenetic mechanisms associated with downregulation of MAL, including loss of heterozygosity (LOH), mutation, and hypermethylation. Thirty-five cell lines of University of Turko squamous cell carcinoma (UT-SCC) series derived from head and neck cancer, including nine pairs from matched primary and metastatic tumors, and 30 pairs of matched primary and metastatic tumor samples were analyzed. Twenty out of 35 (57%) cell lines showed downregulation of MAL expression, whereas no expression was found in 10 cell lines (29%). Considering matched primary and metastatic tumor-derived cell-line pairs, four pairs showed decreased expression only in metastasis-derived cells compared with their primary counterparts. Expression analysis of 21 tissue samples demonstrated decreased or no expression of MAL mRNA in 43% of metastatic tumors compared with matched primary tumors. Relating to mechanisms of downregulation, LOH was observed in 30% of primary tumors and 38% of their metastatic counterparts by a MAL-specific microsatellite marker. Furthermore, we found restoration of MAL mRNA after treatment with demethylating agent (5-aza-2'-deoxycytidine) in 9 (45%) out of 20 cell lines. No mutation was found in UT-SCC cell lines. In conclusion, our findings indicate selective downregulation of MAL expression in metastatic cells, suggesting the MAL gene as a new metastasis-suppressor candidate for head and neck cancer. LOH and hypermethylation appeared to be important mechanisms for inactivation of MAL function.


Subject(s)
Carcinoma, Squamous Cell/metabolism , Head and Neck Neoplasms/metabolism , Membrane Transport Proteins/metabolism , Myelin Proteins/metabolism , Proteolipids/metabolism , Adult , Aged , Aged, 80 and over , Azacitidine/analogs & derivatives , Azacitidine/pharmacology , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Carcinoma, Squamous Cell/genetics , Cell Line, Tumor , Decitabine , Female , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/genetics , Head and Neck Neoplasms/genetics , Heterozygote , Humans , Male , Membrane Transport Proteins/genetics , Middle Aged , Mutation/genetics , Myelin Proteins/genetics , Myelin and Lymphocyte-Associated Proteolipid Proteins , Neoplasm Metastasis/genetics , Neoplasm Metastasis/pathology , Proteolipids/genetics , RNA, Messenger/genetics
12.
Head Neck ; 31(6): 793-801, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19260128

ABSTRACT

BACKGROUND: Although contribution of matrix metalloproteinases in cancer progression and dissemination is now well known, roles of recently discovered metalloproteinases, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), in cancer development and progression remain mostly unknown. METHODS: Here we examined the mRNA expression pattern of 6 members of ADAMTS aggrecanases (1, 4, 5, 8, 9, and 15) in primary head and neck cancer with and without metastasis by real-time reverse transcriptase-polymerase chain reaction. RESULTS: Expression levels of ADAMTS mRNAs were lower in the majority of the primary tumors as compared with the controls. On the other hand, the expression levels of all of the ADAMTS mRNAs except ADAMTS4 were higher in the metastatic foci than in their corresponding primary tumors, which suggest that characteristics of the cancer cell population are different in the primary tumor and metastatic focus. CONCLUSION: Our findings suggest a metastasis model proposing accumulation of a subtype of cancer cells with high metastatic capacity within heterogenous primary tumor cell population.


Subject(s)
ADAM Proteins/metabolism , Biomarkers, Tumor/metabolism , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/secondary , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/pathology , ADAM Proteins/genetics , ADAMTS5 Protein , Aged , Carcinoma, Squamous Cell/mortality , Case-Control Studies , Female , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/mortality , Humans , Male , Middle Aged , Neoplasm Staging , Prognosis , RNA, Messenger/analysis , Reference Values , Reverse Transcriptase Polymerase Chain Reaction , Risk Assessment , Sensitivity and Specificity , Statistics, Nonparametric , Survival Analysis
13.
Arch Otolaryngol Head Neck Surg ; 135(3): 254-60, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19289703

ABSTRACT

OBJECTIVE: To examine the role of TESTIN as a candidate tumor suppressor gene in head and neck carcinogenesis. DESIGN: Mutation and messenger RNA (mRNA) expression analyses. SETTING: Academic research. PATIENTS: Paired normal and tumor samples were obtained from 38 patients with primary head and neck squamous cell carcinoma. MAIN OUTCOME MEASURES: Analysis and comparison of TESTIN gene mRNA expression and its relationship to clinicopathologic variables. RESULTS: Mutation analysis showed a nucleotide and amino acid change in 6 of the 38 tumor samples (16.0%). Semiquantitative mRNA expression analysis of TESTIN revealed a decreased expression in approximately 50% of the tumors compared with their matched normal controls. Interestingly, comparison of clinicopathologic variables to mRNA expression status of TESTIN revealed a significant difference in terms of cancer history (P = .03). Moreover, a higher smoking ratio and a family cancer history were also associated with downregulation of TESTIN, although the difference was not statistically significant (P = .43 and P = .16, respectively). Kaplan-Meier survival analysis demonstrated a worse survival rate among the patients with low TESTIN expression compared with the patients with normal-high TESTIN expression. CONCLUSIONS: Our findings suggest that inactivation of TESTIN is involved in head and neck carcinogenesis through its downregulation. Further studies in various human cancer tissues using a large sample size and in vitro functional studies as well as clinical comparison research studies would give us a better evaluation of TESTIN's role and its possible future application in molecular diagnosis and treatment of different cancer types, including head and neck squamous cell carcinoma.


Subject(s)
Carcinoma, Squamous Cell/genetics , Down-Regulation , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/genetics , Homeodomain Proteins/genetics , RNA, Neoplasm/genetics , Tumor Suppressor Proteins/genetics , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/pathology , Cytoskeletal Proteins , DNA Mutational Analysis , Disease Progression , Female , Head and Neck Neoplasms/mortality , Head and Neck Neoplasms/pathology , Homeodomain Proteins/metabolism , Humans , Japan/epidemiology , LIM Domain Proteins , Male , Middle Aged , Mutation , Prognosis , RNA, Neoplasm/metabolism , RNA-Binding Proteins , Retrospective Studies , Reverse Transcriptase Polymerase Chain Reaction , Survival Rate , Tumor Suppressor Proteins/metabolism
14.
Auris Nasus Larynx ; 36(5): 578-85, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19268507

ABSTRACT

OBJECTIVE: We aimed to define role of tonsillar lymphocytes (TL) and immune cross-reactivity between bacterial-HSP65 and human-HSP60 in Pustulosis palmaris et plantaris (PPP), an intractable chronic disease characterized with pustules and cornification of palms and soles. METHODS: Two sets of crossover trials were designed by employing SCID mice model. In the first trial, mice were transplanted with tonsillar lymphocytes and skin-grafts from PPP patients (TL group). In the second trial, mice were transplanted with tonsillar lymphocytes from PPP patients and injected with recombinant human HSP60. Control groups were designed for each step. Comparisons were performed for immunologic analyses including infiltration of CD4+ lymphocytes in skin-grafts by immunostaining, and levels of anti-HSP65-IgG and cytokines in mice sera by enzyme-linked immunosorbent assay (ELISA). RESULTS: In TL group, infiltration of CD4+ lymphocytes in skin-grafts were significantly higher than mice transplanted with blood lymphocytes (p<0.05), while anti-HSP65-IgG levels in sera showed non-significant tendency to increase in the TL group. CD4+ cells and anti-HSP65-IgG levels were also well-correlated with each other in TL group (p<0.01). Besides, anti-HSP65-IgG levels were significantly correlated with cytokine levels (IL-6, IFN-gamma) in mice sera (p<0.01). We found strong expression of HSP60 in PPP lesions. Finally, HSP60-stimulation in mice transplanted with TL from PPP patients induced significantly higher anti-HSP65-IgG levels in serum compared to control groups including mice without HSP60-stimulation or peripheral blood lymphocytes-transplanted mice or transplanted with TL from control patients (p<0.05). CONCLUSION: Our results indicate the pathogenic role of TL and immune cross-reaction between human-HSP60 and bacterial-HSP65 in PPP.


Subject(s)
Bacterial Proteins/immunology , Chaperonin 60/immunology , Cross Reactions , Heat-Shock Proteins/immunology , Lymphocytes/immunology , Palatine Tonsil/immunology , Psoriasis/immunology , Adult , Animals , Antibodies/blood , Biopsy , CD4 Antigens/metabolism , CD4 Lymphocyte Count , Chaperonin 60/metabolism , Chimera , Cross-Over Studies , Cytokines/blood , Epithelium/metabolism , Epitopes , Female , Humans , Immunoglobulin G/blood , Male , Mice , Mice, SCID/blood , Middle Aged , Palatine Tonsil/pathology , Psoriasis/metabolism , Psoriasis/pathology , Skin/metabolism , Skin/pathology , Skin Transplantation , Tonsillectomy , Transplantation, Heterologous
15.
Eur J Pediatr ; 168(11): 1365-72, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19221788

ABSTRACT

Human Bocavirus (HBoV) as a newly discovered parvovirus has been commonly detected in respiratory tract infections. However, its role in acute otitis media (AOM) has not been well studied. We examined HBoV in Japanese children with AOM and evaluated the virus prevalence together with clinical manifestations and bacterial findings. Overall, 222 nasopharyngeal swabs and 176 middle ear fluids (MEF) samples were collected from 222 children with AOM (median age, 19 months) between May 2006 and April 2007. HBoV detection was performed by PCR and bacterial isolation by standard culture methods. HBoV was found in the nasopharyngeal aspirates of 14 children (6.3%) and in the MEF of six children (2.7%). When HBoV detection results were evaluated with clinical characteristics of children, resolution time of AOM was significantly longer (p=0.04), and rate of fever symptom was also higher in HBoV-positive group (p=0.04). Furthermore, we found positive correlation between detection of HBoV and Streptococcus pneumoniae in the MEF (p=0.004). Nevertheless, nasopharyngeal proportion of S. pneumoniae was similar between virus positive and negative groups. Furthermore, S. pneumoniae was detected as a single pathogen in all MEF of HBoV-positive cases but one, while it presents mixed with other pathogenic bacteria in nasopharynx. In conclusion, HBoV may worsen the clinical symptoms and prolong the clinical outcome of AOM in pediatric population. Finally, HBoV may prime the secondary bacterial infection in the middle ear in favor of S. pneumoniae.


Subject(s)
Human bocavirus/isolation & purification , Otitis Media/diagnosis , Otitis Media/virology , Parvoviridae Infections/diagnosis , Acute Disease , Adolescent , Anti-Bacterial Agents/therapeutic use , Bacterial Infections/diagnosis , Child , Child, Preschool , Ear, Middle/microbiology , Ear, Middle/virology , Female , Humans , Infant , Japan/epidemiology , Male , Middle Ear Ventilation , Nasopharynx/microbiology , Nasopharynx/virology , Otitis Media/epidemiology , Otitis Media/microbiology , Otitis Media/therapy , Parvoviridae Infections/epidemiology , Parvoviridae Infections/therapy , Parvoviridae Infections/virology , Polymerase Chain Reaction , Prevalence , Retrospective Studies , Severity of Illness Index , Streptococcus pneumoniae/isolation & purification , Treatment Outcome
16.
J Oral Pathol Med ; 38(1): 94-8, 2009 Jan.
Article in English | MEDLINE | ID: mdl-18363675

ABSTRACT

BACKGROUND: Squamous cell carcinoma is the most common cancer type of the oral cavity and approximately 50% of the patients succumb to the disease. Unfortunately, few are known about the molecular mechanisms involving in the formation of oral squamous cell carcinoma (OSCC). Recently, it has been reported that 1p36 chromosomal region is deleted in various cancer types and is suspected to harbor various tumor suppressor genes (TSGs). However, limited studies exist on genetics alteration on 1p36 in OSCC and the responsible TSG remained unidentified. METHODS: To investigate area susceptible to harbor TSG(s) involved in OSCC on 1p36 region, paired normal and tumor tissues of 27 patients with diagnosis of OSCC have been analyzed for loss of heterozygosity (LOH) using nine microsatellite markers based on recent gene mapping. RESULTS: LOH was found at least in one locus in 85% of the cases (23 of 27). Interestingly, microsatellite instability was also found in 7% (two of 27) of the cases analyzed. The higher LOH frequencies were found with the markers D1S243 (25%), D1S468 (22%), D1S450 (25%), D1S228 (38%), D1S199 (28%), and D1S1676 (23%). CONCLUSIONS: Three preferentially deleted regions have been identified in OSCC: region 1 (D1S468-D1S243), region 2 (D1S450-D1S228), and region 3 (D1S199-D1S1676). Multiple candidate TSGs, such as RIZ1, p73, UBE4B, Rap1GAP, EPHB2, and RUNX3, are located in these three areas. The data obtained in this study can be used for further functional analysis of these genes involved in OSCC carcinogenesis.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 1/genetics , Gene Deletion , Mouth Neoplasms/genetics , Apoptosis/genetics , Chromosome Mapping , Core Binding Factor Alpha 3 Subunit/genetics , DNA-Binding Proteins/genetics , GTPase-Activating Proteins/genetics , Gene Frequency/genetics , Genes, Tumor Suppressor , Histone-Lysine N-Methyltransferase , Humans , Loss of Heterozygosity/genetics , Microsatellite Instability , Microsatellite Repeats/genetics , Nuclear Proteins/genetics , Receptor, EphB2/genetics , Transcription Factors/genetics , Tumor Protein p73 , Tumor Suppressor Proteins/genetics , Ubiquitin-Protein Ligase Complexes/genetics , Ubiquitin-Protein Ligases
17.
J Cancer Res Clin Oncol ; 135(5): 703-13, 2009 May.
Article in English | MEDLINE | ID: mdl-18998165

ABSTRACT

BACKGROUND: Loss of heterozygosity (LOH) in the ING family members has been shown in head and neck squamous cell carcinoma (HNSCC) except for ING2. Like all the other members of ING family, ING2, which is located at chromosome 4q35.1, is a promising tumor suppressor gene (TSG). In this study, we performed LOH analysis of ING2 in HNSCC and compared it with clinicopathological variables. MATERIALS AND METHODS: We performed LOH analysis in DNAs from 80 paired of normal and HNSCC tissues, using a specifically designed microsatellite marker on chromosome 4q35.1, which detects allelic loss of ING2. TP53 mutation analysis and its relationship with ING2 chromosomal deletion were also performed in available 68 of the samples. The correlation between LOH status and clinicopathological characteristics was evaluated by using statistical methods. The overall survival (OS) and disease free survival (DFS) were also determined. RESULTS: LOH was detected in 54.6% (30/55) of the informative samples. Statistical significance was obtained between LOH and tumor (T) stage (P = 0.02), application of radiotherapy and chemotherapy. Positive node status (N) appeared to be the only independent prognostic factor for both OS (P = 0.031) and DFS (P = 0.044). CONCLUSIONS: Our study showed allelic loss of 4q35.1 in HNSCC. The high percentage of LOH suggests ING2 as a candidate TSG in HNSCC. High LOH frequency was statistically associated with advanced T stage, suggesting that ING2 LOH might occur in late stages during HNSCC progression.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 4 , Gene Deletion , Head and Neck Neoplasms/genetics , Homeodomain Proteins/genetics , Loss of Heterozygosity , Receptors, Cytoplasmic and Nuclear/genetics , Tumor Suppressor Proteins/genetics , Aged , Carcinoma, Squamous Cell/mortality , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/surgery , Chromosome Mapping , Female , Genes, Tumor Suppressor , Genes, p53 , Head and Neck Neoplasms/pathology , Head and Neck Neoplasms/surgery , Humans , Male , Microsatellite Repeats , Middle Aged , Mutation , Neoplasm Staging/mortality , Survival Analysis , Survivors
18.
Cancer Sci ; 99(8): 1589-94, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18754871

ABSTRACT

The epidermal growth factor receptor (EGFR)-RAS-RAF-mitogen-activated protein kinase signaling cascade is an important pathway in cancer development and recent reports show that EGFR and its downstream signaling molecules are mutated in a number of cancers. We have analyzed 91 Japanese head and neck squamous cell carcinomas (HNSCC) and 12 HNSCC cell lines for mutations in EGFR, ErbB2, and K-ras. Exons encoding the hot-spot regions in the tyrosine kinase domain of both EGFR (exons 18, 19, and 21) and ErbB2 (exons 18-23), as well as exons 1 and 2 of K-ras were amplified by polymerase chain reaction and sequenced directly. EGFR expression was also analyzed in 65 HNSCC patients using immunohistochemistry. Only one silent mutation, C836T, was found in exon 21 of EGFR in the UT-SCC-16A cell line and its corresponding metastasic cell line UT-SCC-16B. No other mutation was found in EGFR, ErbB2, or K-ras. All tumors showed EGFR expression. In 21 (32%) tumors, EGFR was expressed weakly (+1). In 27 (42%) tumors it was expressed (+2) moderately, and in 17 (26%) tumors high expression (+3) was detected. Overexpression (+2, +3) was found in 44 tumors (68%). A worse tumor differentiation and a positive nodal stage were significantly associated with EGFR overexpression (P = 0.02, P = 0.032, respectively). Similar to patients from western ethnicity, mutations are absent or rare in Japanese HNSCC. Protein overexpression rather than mutation might be responsible for activation of the EGFR pathway in HNSCC.


Subject(s)
ErbB Receptors/biosynthesis , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/metabolism , Mutation , Neoplasms, Squamous Cell/metabolism , Aged , Cell Line, Tumor , Female , Gene Expression , Genes, erbB-2/genetics , Genes, ras/genetics , Head and Neck Neoplasms/genetics , Humans , Immunohistochemistry , Japan , Male , Middle Aged , Neoplasms, Squamous Cell/genetics , Polymerase Chain Reaction
19.
J Cancer Res Clin Oncol ; 134(12): 1267-76, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18521630

ABSTRACT

PURPOSE AND METHODS: Loss of heterozygosity (LOH) in a chromosomal location indicates the presence of an inactivated tumor suppressor gene (TSG). Inactivation of TSG has a functional role in the tumorigenesis of head and neck squamous cell carcinoma (HNSCC). Based on the recent evidences of a putative TSG on chromosome 14, we examined LOH on chromosome 14q using eight polymorphic microsatellite markers in 50 cases of HNSCCs. RESULTS: Three regions were detected to have a high LOH rate which included 14q21.2-22.3 (42.5%), 14q31 (55%), and 14q32.1 (37%). The correlation between LOH and clinicopathological findings was investigated through statistical analyses. A strong correlation was observed between the highest LOH marker and the overall and disease-free survival. CONCLUSIONS: The results suggest that the distal part of chromosome 14 may host a TSG that may lead to the development and/or progression of HNSCCs. Several genes such as CHES1, BMP4, SAV, and PNN have arisen as candidate tumor suppressors in the region.


Subject(s)
Carcinoma, Squamous Cell/genetics , Chromosomes, Human, Pair 14/genetics , Head and Neck Neoplasms/genetics , Loss of Heterozygosity , Aged , Carcinoma, Squamous Cell/pathology , Case-Control Studies , Chromosome Deletion , Chromosome Mapping , Female , Genes, Tumor Suppressor , Head and Neck Neoplasms/pathology , Humans , Male , Microsatellite Repeats , Middle Aged , Prognosis , Survival Rate
20.
Oncol Rep ; 19(5): 1079-84, 2008 May.
Article in English | MEDLINE | ID: mdl-18425361

ABSTRACT

EphA2 is a 130-kDa transmembrane protein primarily found in adult human epithelial cells and is a member of one of the largest receptor tyrosine kinases. It is located on 1p36.1, a genetic hot spot in cancer. EphA2 overexpression has been observed in aggressive solid tumors and its potential role in tumorigenesis, which includes cell growth, survival, migration and angiogenesis have been reported. However, the role of EphA2 remains unknown in head and neck cancer. In this study, we investigated the genetic profile of EphA2 in primary head and neck squamous cell carcinoma (HNSCC) by determining mRNA level, status of loss of heterozygosity and protein expression. mRNA expression was also correlated with clinicopathological data. Infrequent loss of heterozygosity (20%) was observed, though a 10-fold increase of mRNA expression in tumors compared to normal tissues was noted. A significant number of samples with normal to high mRNA expression was observed among patients with regional metastasis, with T3-T4 tumor size and with moderate to poor differentiation. However, statistical studies did not show any correlation between mRNA expression and any of the clinicopathological parameters. Tumor cells expressed EphA2 protein, but only weakly. These results suggest that EphA2 might be involved in the early development of HNSCC although not directly responsible for its progression.


Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/metabolism , Gene Expression Regulation, Neoplastic , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/metabolism , Loss of Heterozygosity , RNA, Messenger/metabolism , Receptor, EphA2/biosynthesis , Receptor, EphA2/genetics , Aged , Female , Gene Expression Profiling , Humans , Ligands , Male , Middle Aged , Models, Genetic , Neoplasm Metastasis
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