RESUMEN
The aim of this study was to compare the expressions of basal lamina (BL) collagen IV α chains and matrix metalloproteinases (MMP)-2 and MMP-9 in oral dysplasia (OED) and invasive carcinoma. Ten cases each of OEDs, carcinomas-in situ and oral squamous cell carcinomas (OSCCs) were examined by immunohistochemistry. Another 5 cases, each of normal and hyperplastic oral mucosa, served as controls. Results showed that α1(IV)/α2(IV) and α5(IV)/α6(IV) chains were intact in BLs of control and OEDs. In BLs of carcinoma-in situ, α1(IV)/α2(IV) chains preceded α5(IV)/α6(IV) chains in showing incipient signs of disruption. OSCCs exhibited varying degrees of collagen α(IV) chain degradation. MMP-2 and MMP-9 were absent in controls and OED, but weakly detectable in carcinoma-in situ. In OSCC, these proteolytic enzymes were expressed in areas corresponding to collagen α(IV) chain loss. Enzymatic activity was enhanced in higher grade OSCC, and along the tumor advancing front. Overall the present findings suggest that loss of BL collagen α(IV) chains coincided with gain of expression for MMP-2 and MMP-9, and that these protein alterations are crucial events during progression from OED to OSCC.
Asunto(s)
Carcinoma/enzimología , Colágeno Tipo IV/química , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Neoplasias de la Boca/enzimología , Adulto , Anciano , Anciano de 80 o más Años , Membrana Basal/enzimología , Carcinoma in Situ , Estudios de Casos y Controles , Colágeno Tipo IV/metabolismo , Progresión de la Enfermedad , Femenino , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica/métodos , Masculino , Persona de Mediana Edad , Mucosa Bucal/patología , Invasividad NeoplásicaRESUMEN
Lymph node metastasis is considered a factor in determining the prognosis of squamous cell carcinoma (SCC). Both oral and cervical SCC tumor cells prefer lymph vessels as the route of metastasis. D2-40 is a specific marker of lymphatic endothelial cells. This study clarifies the distribution and characteristics of lymphatic vessels in oral and cervical SCCs. Immunohistochemistry was performed in 20 oral and 20 cervical SCCs (10 non-metastatic and 10 metastatic to lymph nodes) using D2-40, CD31, CD34, CD105 and double staining with D2-40 and keratin. Lymphatic vessel density (LVD) was also determined morphologically. Results showed that lymphatic vessels in both types of SCCs were distributed mainly at the superficial region beneath the epithelium. The LVD in each tumor was significantly higher compared to the corresponding normal mucosa. Moreover, the LVD in lymph node metastasis in each tumor was significantly higher compared to their non-metastatic counterparts. Cancer cell invasion was observed in the lymphatic vessels suggesting the existence of lymph node involvement during metastasis. The new lymphatic vessels that proliferated around the cancer nests in both SCCs have endothelial cell characteristics inferred to be associated with early lymphatic development and initial dissemination of cancer cells.
RESUMEN
Biological apatites are characterized by the presence of minor constituents such as magnesium (Mg), chloride (Cl), or fluoride (F) ions. These ions affect cell proliferation and osteoblastic differentiation during bone tissue formation. F-substituted apatites are being explored as potential bonegraft materials. The aim of the present study is to investigate the mechanism of bone formation induced by fluoride-substituted apatite (FAp) by analyzing the effect of FAp on the process of in vivo bone formation. FAps containing different F concentrations (l-FAp: 0.48 wt%, m-FAp: 0.91 wt%, h-FAp: 2.23 wt%) and calcium-deficient apatite (CDA), as positive control, were implanted in rat tibia and bone formation was evaluated by histological examination, immuhistochemistry, in situ hybridization and tartrate-resistant acid phosphatase examinations. The results showed that l-FAp, m-FAp, h-FAp, and CDA biomaterials allowed migration of macrophages, attachment, proliferation, and phenotypic expression of bone cells leading to new bone formation in direct apposition to the particles. However, the l-FAp preparation allowed faster bone conduction compared to the other experimental materials. These results suggest that FAp with low F concentration may be an efficient bonegraft material for dental and medical application.
Asunto(s)
Apatitas/administración & dosificación , Apatitas/química , Sustitutos de Huesos/química , Calcio/química , Fluoruros/administración & dosificación , Fluoruros/química , Osteogénesis/efectos de los fármacos , Animales , Huesos/efectos de los fármacos , Calcio/administración & dosificación , Diferenciación Celular/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Ratas , Tibia/efectos de los fármacos , Tibia/metabolismoRESUMEN
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.
Asunto(s)
Ameloblastoma/genética , Proteínas de Ciclo Celular/genética , Proteínas de Homeodominio/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Neoplasias Maxilomandibulares/genética , Pérdida de Heterocigocidad , Proteínas Nucleares/genética , Transactivadores/genética , Proteínas Supresoras de Tumor/genética , Adolescente , Adulto , Anciano , Ameloblastoma/patología , Niño , Femenino , Genes Supresores de Tumor , Humanos , Proteína Inhibidora del Crecimiento 1 , Masculino , Persona de Mediana EdadRESUMEN
Titanium and hydroxyapatite (HA) are widely used as biomaterials for dental and medical applications. HA-coated titanium implants have excellent biocompatibility and mechanical properties. However, the adherence of HA film formed on titanium substrate is weak because of the lack of chemical interaction between HA and titanium. A solution to this problem is to form an intermediate film on titanium substrate, which provide excellent adherence to both titanium substrate and HA. We developed a novel biomaterial called calcium titanate-amorphous carbon (CaTiO(3)-aC) coating prepared by modified thermal decomposition method. The purpose of this study was to evaluate the effect of CaTiO(3)-aC and HA coating (positive control), and Ti (negative control) on osteoblastic (MT3T3-E1) cell responses. An increased cellular proliferation was observed in CaTiO(3)-aC coating compared to HA coating. The maximum expressions of ALP activity, Col I and ALP mRNA were higher and achieved in shorter period of time in CaTiO(3)-aC coating compared to others. These results demonstrated that CaTiO(3)-aC promoted better cell attachment, cellular proliferation, and osteoblastic differentiation compared with HA. In conclusion, we suggested that CaTiO(3)-aC could be considered as an important candidate as a coating material.
Asunto(s)
Materiales Biocompatibles Revestidos/química , Osteoblastos/citología , Titanio/química , Células 3T3 , Animales , Materiales Biocompatibles/química , Sustitutos de Huesos , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Calor , Ratones , Microscopía Electrónica de Rastreo/métodos , Microscopía Electrónica de Transmisión/métodos , PolvosRESUMEN
Progenitor cells play an important biological role in tooth and bone formation, and previous analyses during bone and dentine induction have indicated that they may be a good alternative for tissue engineering. Thus, to clarify the influence of the microenvironment on protein and gene expression, MDPC-23 cells (mouse dental papilla cell line) and KUSA/A1 cells (bone marrow stromal cell line) were used, both in vitro cell culture and in intra-abdominal diffusion chambers implanted in 4-week-old male immunodefficient mice (SCID mice). Our results indicate that KUSA/A1 cells differentiated into osteoblast-like cells and induced bone tissue inside the chamber, whereas, MDPC-23 showed odontoblast-like characteristics but with a low ability to induce dentin formation. This study shows that MDPC-23 cells are especial cells, which possess morphological and functional characteristics of odontoblast-like cells expressing dentin sialophosphoprotein in vivo. In contrast, dentin sialophosphoprotein gene and protein expression was not detected in both cell lines in vitro. The intra-abdominal diffusion chamber appears as an interesting experimental model for studying phenotypic expression of dental pulp cells in vivo.
Asunto(s)
Regeneración Ósea/fisiología , Diferenciación Celular/fisiología , Odontoblastos/citología , Odontogénesis/fisiología , Osteoblastos/citología , Animales , Regeneración Ósea/genética , Huesos/citología , Huesos/metabolismo , Diferenciación Celular/genética , Línea Celular , Colágeno Tipo I/biosíntesis , Colágeno Tipo I/genética , Papila Dental/citología , Papila Dental/metabolismo , Cámaras de Difusión de Cultivos/métodos , Expresión Génica , Masculino , Ratones , Ratones SCID , Odontoblastos/metabolismo , Odontogénesis/genética , Osteoblastos/metabolismo , Osteocalcina/biosíntesis , Osteocalcina/genética , Osteonectina/biosíntesis , Osteonectina/genética , Osteopontina/biosíntesis , Osteopontina/genética , Biosíntesis de Proteínas , Sialoglicoproteínas/biosíntesis , Sialoglicoproteínas/genéticaRESUMEN
PURPOSE: Mucoepidermoid carcinoma (MEC) is the most frequently detected primary malignancy of the salivary gland and is characterized by a marked variation in prognosis. In the present study, we investigated the prognostic significance of p27Kip1, Ki-67, and CRTC1 (also called MECT1, TORC1, and WAMTP1)-MAML2 fusion in MEC. MATERIALS AND METHODS: MEC cases (n = 101) were examined for p27Kip1 and Ki-67 expression using immunohistochemistry and for CRTC1-MAML2 fusion transcript using reverse transcriptase-polymerase chain reaction. RESULTS: p27Kip1, Ki-67, and the CRTC1-MAML2 fusion transcript were expressed in 71, 31, and 34 of the 101 cases, respectively. p27Kip1 and CRTC1-MAML2 fusion were associated with favorable clinicopathologic tumor features and Ki-67 with aggressive clinicopathologic features. Multivariate survival analyses were performed that included the following 10 clinicopathologic factors: age, gender, tumor site, tumor size, nodal metastasis, clinical stage, histologic grade, p27 expression, Ki-67 expression, and CRTC1-MAML2 fusion. For disease-free survival, only p27Kip1 expression was significant as an independent prognostic factor. For overall survival, p27Kip1 expression, CRTC1-MAML2 fusion, and tumor size were significant. In each analysis, p27Kip1 and CRTC1-MAML2 fusion were independent of the clinical stage. Ki-67 expression was not selected in either multivariate analysis. CONCLUSIONS: p27Kip1 and CRTC1-MAML2 fusion were associated with favorable clinicopathologic tumor features, and both were useful in predicting the overall survival of patients with MEC. For disease-free survival, p27Kip1 might be the most useful prognostic factor. In contrast, Ki-67 might not be a very powerful prognostic indicator for either survival point.
Asunto(s)
Biomarcadores de Tumor/biosíntesis , Carcinoma Mucoepidermoide/metabolismo , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/biosíntesis , Antígeno Ki-67/biosíntesis , Proteínas de Fusión Oncogénica/biosíntesis , Neoplasias de las Glándulas Salivales/metabolismo , Factores de Edad , Biomarcadores de Tumor/genética , Carcinoma Mucoepidermoide/mortalidad , Carcinoma Mucoepidermoide/patología , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Antígeno Ki-67/genética , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Estadificación de Neoplasias , Proteínas de Fusión Oncogénica/genética , Pronóstico , ARN Neoplásico/análisis , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias de las Glándulas Salivales/mortalidad , Neoplasias de las Glándulas Salivales/patología , Factores Sexuales , Tasa de SupervivenciaRESUMEN
BACKGROUND: Although it is well known that olfactory receptor neurons differentiated from stem cells have an unusual lifelong ability to continue neurogenesis, effective treatment of patients with sensorineural olfactory loss is still lacking. The purpose of this study was to determine whether intranasal prolactin (PRL) is an effective inducer of proliferation in the olfactory epithelium (OE). METHODS: The OE of mice treated with PRL for 6 consecutive days and of pregnant mice on gestation day (GD) 7 was investigated by immunohistochemical study using antibodies to bromodeoxyuridine. RESULTS: The number of bromodeoxyuridine -labeled cells in the OE increased significantly in PRL-treated female mice and GD 7 mice (but not PRL-treated male mice) compared with controls. CONCLUSION: Pregnancy may induce proliferation in the OE, and administration of intranasal PRL may have almost the same effect as pregnancy on the OE of female mice.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Mucosa Olfatoria/patología , Prolactina/metabolismo , Administración Intranasal , Animales , Bromodesoxiuridina , Células Cultivadas , Femenino , Pérdida Auditiva Sensorineural/metabolismo , Pérdida Auditiva Sensorineural/patología , Pérdida Auditiva Sensorineural/prevención & control , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos BALB C , Mucosa Olfatoria/metabolismo , Embarazo , Prolactina/farmacología , Factores SexualesRESUMEN
Progenitor cells play an important biological role in tooth and bone formation, and previous analyses during bone and dentine induction have indicated that they may be a good alternative for tissue engineering. Thus, to clarify the influence of the microenvironment on protein and gene expression, MDPC-23 cells (mouse dental papilla cell line) and KUSA/A1 cells (bone marrow stromal cell line) were used, both in vitro cell culture and in intra-abdominal diffusion chambers implanted in 4-week-old male immunodefficient mice (SCID mice). Our results indicate that KUSA/A1 cells differentiated into osteoblast-like cells and induced bone tissue inside the chamber, whereas, MDPC-23 showed odontoblast-like characteristics but with a low ability to induce dentin formation. This study shows that MDPC-23 cells are especial cel ls, which possess morphological and functional characteristics of odontoblast-like cells expressing dentin sialophosphoprotein in vivo. In contrast, dentin sialophosphoprotein gene and protein expression was not detected in both cell lines in vitro. The intra-abdominal diffusion chamber appears as an interesting experimental model for studying phenotypic expression of dental pulp cells in vivo.
Asunto(s)
Masculino , Animales , Ratones , Colágeno Tipo I/biosíntesis , Colágeno Tipo I/genética , Odontoblastos/citología , Odontoblastos/metabolismo , Regeneración Ósea/fisiología , Regeneración Ósea/genética , Sialoglicoproteínas/biosíntesis , Sialoglicoproteínas/genética , Huesos/citología , Huesos/metabolismo , Ratones SCID , Osteocalcina/biosíntesis , Osteocalcina/genética , Osteonectina/biosíntesis , Osteonectina/genética , Osteopontina/biosíntesis , Osteopontina/genéticaRESUMEN
Progenitor cells play an important biological role in tooth and bone formation, and previous analyses during bone and dentine induction have indicated that they may be a good alternative for tissue engineering. Thus, to clarify the influence of the microenvironment on protein and gene expression, MDPC-23 cells (mouse dental papilla cell line) and KUSA/A1 cells (bone marrow stromal cell line) were used, both in vitro cell culture and in intra-abdominal diffusion chambers implanted in 4-week-old male immunodefficient mice (SCID mice). Our results indicate that KUSA/A1 cells differentiated into osteoblast-like cells and induced bone tissue inside the chamber, whereas, MDPC-23 showed odontoblast-like characteristics but with a low ability to induce dentin formation. This study shows that MDPC-23 cells are especial cel ls, which possess morphological and functional characteristics of odontoblast-like cells expressing dentin sialophosphoprotein in vivo. In contrast, dentin sialophosphoprotein gene and protein expression was not detected in both cell lines in vitro. The intra-abdominal diffusion chamber appears as an interesting experimental model for studying phenotypic expression of dental pulp cells in vivo.(AU)
Asunto(s)
Masculino , Animales , Ratones , Regeneración Ósea/genética , Regeneración Ósea/fisiología , Colágeno Tipo I/biosíntesis , Colágeno Tipo I/genética , Odontoblastos/citología , Odontoblastos/metabolismo , Sialoglicoproteínas/biosíntesis , Sialoglicoproteínas/genética , Huesos/citología , Huesos/metabolismo , Osteocalcina/biosíntesis , Osteocalcina/genética , Osteonectina/biosíntesis , Osteonectina/genética , Osteopontina/biosíntesis , Osteopontina/genética , Ratones SCIDRESUMEN
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.
Asunto(s)
Carcinoma de Células Escamosas/genética , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/genética , Proteínas de Homeodominio/genética , ARN Neoplásico/genética , Proteínas Supresoras de Tumor/genética , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Células Escamosas/mortalidad , Carcinoma de Células Escamosas/patología , Proteínas del Citoesqueleto , Análisis Mutacional de ADN , Progresión de la Enfermedad , Femenino , Neoplasias de Cabeza y Cuello/mortalidad , Neoplasias de Cabeza y Cuello/patología , Proteínas de Homeodominio/metabolismo , Humanos , Japón/epidemiología , Proteínas con Dominio LIM , Masculino , Persona de Mediana Edad , Mutación , Pronóstico , ARN Neoplásico/metabolismo , Proteínas de Unión al ARN , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tasa de Supervivencia , Proteínas Supresoras de Tumor/metabolismoRESUMEN
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.
Asunto(s)
Carcinoma de Células Escamosas/genética , Cromosomas Humanos Par 1/genética , Eliminación de Gen , Neoplasias de la Boca/genética , Apoptosis/genética , Mapeo Cromosómico , Subunidad alfa 3 del Factor de Unión al Sitio Principal/genética , Proteínas de Unión al ADN/genética , Proteínas Activadoras de GTPasa/genética , Frecuencia de los Genes/genética , Genes Supresores de Tumor , N-Metiltransferasa de Histona-Lisina , Humanos , Pérdida de Heterocigocidad/genética , Inestabilidad de Microsatélites , Repeticiones de Microsatélite/genética , Proteínas Nucleares/genética , Receptor EphB2/genética , Factores de Transcripción/genética , Proteína Tumoral p73 , Proteínas Supresoras de Tumor/genética , Complejos de Ubiquitina-Proteína Ligasa/genética , Ubiquitina-Proteína LigasasRESUMEN
To investigate whether bone marrow-derived cells (BMC) would migrate and engraft into the sensory epithelium of the inner ear, BMC of green fluorescence protein (GFP) mice were transplanted into lethally irradiated recipient mice. Then the recipient mice were treated with streptomycin and immunohistochemical staining was performed to evaluate the migration and engraftment of donor BMC into the sensory epithelium of the inner ear. Immunohistochemical staining for GFP was found initially in the vascular epithelium and oral mucosa but not in the sensory epithelium of the inner ear. In the case of mouse, BMC may not migrate and be engrafted into the sensory epithelium of the inner ear.
Asunto(s)
Trasplante de Médula Ósea/métodos , Estreptomicina/farmacología , Animales , Trasplante de Médula Ósea/efectos adversos , Movimiento Celular/fisiología , Modelos Animales de Enfermedad , Oído Interno/metabolismo , Oído Interno/patología , Oído Interno/cirugía , Femenino , Rechazo de Injerto , Supervivencia de Injerto , Proteínas Fluorescentes Verdes/biosíntesis , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas del Tejido Nervioso/metabolismo , Distribución Aleatoria , Factores de Riesgo , Sensibilidad y Especificidad , Recolección de Tejidos y Órganos/métodosRESUMEN
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.
Asunto(s)
Carcinoma de Células Escamosas/genética , Cromosomas Humanos Par 4 , Eliminación de Gen , Neoplasias de Cabeza y Cuello/genética , Proteínas de Homeodominio/genética , Pérdida de Heterocigocidad , Receptores Citoplasmáticos y Nucleares/genética , Proteínas Supresoras de Tumor/genética , Anciano , Carcinoma de Células Escamosas/mortalidad , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/cirugía , Mapeo Cromosómico , Femenino , Genes Supresores de Tumor , Genes p53 , Neoplasias de Cabeza y Cuello/patología , Neoplasias de Cabeza y Cuello/cirugía , Humanos , Masculino , Repeticiones de Microsatélite , Persona de Mediana Edad , Mutación , Estadificación de Neoplasias/mortalidad , Análisis de Supervivencia , SobrevivientesRESUMEN
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.
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Receptores ErbB/biosíntesis , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/metabolismo , Mutación , Neoplasias de Células Escamosas/metabolismo , Anciano , Línea Celular Tumoral , Femenino , Expresión Génica , Genes erbB-2/genética , Genes ras/genética , Neoplasias de Cabeza y Cuello/genética , Humanos , Inmunohistoquímica , Japón , Masculino , Persona de Mediana Edad , Neoplasias de Células Escamosas/genética , Reacción en Cadena de la PolimerasaRESUMEN
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.
Asunto(s)
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/genética , Neoplasias de Cabeza y Cuello/metabolismo , Pérdida de Heterocigocidad , ARN Mensajero/metabolismo , Receptor EphA2/biosíntesis , Receptor EphA2/genética , Anciano , Femenino , Perfilación de la Expresión Génica , Humanos , Ligandos , Masculino , Persona de Mediana Edad , Modelos Genéticos , Metástasis de la NeoplasiaRESUMEN
Oral mucosal melanoma is an aggressive neoplasm with poor prognosis. Heparanase is an endo-beta-d-glucuronidase, which cleaves heparan sulphate chains. The vascular endothelial growth factor (VEGF) is the most potent angiogenic mitogen and interaction with its receptor (VEGFR) has been associated with angiogenesis. We investigated the expression of these molecules in the progression of oral mucosal melanoma. Immunohistochemistry was carried out in 15 oral melanotic macules and 19 oral melanomas using heparanase, VEGF, VEGFR-2, CD34 and Ki-67. Microvessel density was determined and subjected to statistical analysis. Heparanase and VEGFR-2 were not expressed in the oral melanotic macule. Atypical melanocytes and melanoma cells expressed heparanase, VEGF and VEGFR-2. An intense expression was noted in the early invasive phase, which marks the crucial transition from in situ to the invasive phase. In the invasive component, heparanase was intense but selective in the invasive fronts and at the periphery of nests unlike the extensive expression of VEGF and VEGFR-2. However, hot spots were only observed at the periphery of the nests. In conclusion, melanoma cells expressed heparanase, VEGF and VEGFR-2. The coexpression of these molecules in atypical melanocytes and melanoma cells suggests their function in cell migration and invasion. Moreover, the intense expression in the crucial transition from in situ to the invasive phase suggests their role in the progression of the tumor. The role of VEGF and VEGFR-2 in angiogenesis was evident only at the periphery of the nests in the invasive components.
Asunto(s)
Glucuronidasa/metabolismo , Melanoma/patología , Mucosa Bucal , Neoplasias de la Boca/patología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Progresión de la Enfermedad , Humanos , Melanoma/irrigación sanguínea , Melanoma/metabolismo , Neoplasias de la Boca/irrigación sanguínea , Neoplasias de la Boca/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Although many clinical and pathological prognostic factors such as tumor stage and lymph-node involvement have been described, to date no reliable or clinically applicable marker or tumor aggressiveness has been identified for head and neck cancer. In an attempt to identify such a molecular prognostic marker, we analyzed the mRNA expression status of ING3 by quantitative reverse transcription-polymerase chain reaction. We also examined p53 mutation status and investigated its relationship with ING3, as well its clinicopathological characteristics. About half of the 71 tumor samples demonstrated downregulation of ING3 compared to their matched normal counterparts. Although most clinicopathological variables were not significantly related to ING3 downregulation or p53 mutation status, a significant relationship was detected in terms of overall survival between the cases with low and normal to high ING3 expression. At 5 years follow up, approximately 60% of the patients with normal to high ING3 expression survived, whereas this was 35% in the patients with low ING3 expression. Multivariate analysis also showed downregulation of ING3 as an independent prognostic factor for poor overall survival. These results reveal that ING3 would function as a potential tumor suppressor molecule and that low levels of ING3 may indicate an aggressive nature of head and neck cancer.
Asunto(s)
Biomarcadores de Tumor/genética , Regulación hacia Abajo , Neoplasias de Cabeza y Cuello/diagnóstico , Proteínas de Homeodominio/genética , Transactivadores/genética , Biomarcadores de Tumor/metabolismo , Estudios de Seguimiento , Genes Supresores de Tumor , Genes p53 , Neoplasias de Cabeza y Cuello/genética , Neoplasias de Cabeza y Cuello/patología , Proteínas de Homeodominio/metabolismo , Humanos , Mutación , Pronóstico , ARN Mensajero/metabolismo , Análisis de Supervivencia , Transactivadores/metabolismo , Proteínas Supresoras de TumorRESUMEN
C-kit is a trans-membrane receptor tyrosine kinase (RTK) encoded by the proto-oncogene KIT located at 4q11-12. Gain-of-function mutations arising to c-kit activation independent of its ligand were observed in various tumors related to germ cells, mast cells, and interstitial cells of Cajal. C-kit also participates in melanocyte development; hence, its involvement in oral mucosal melanoma (OMM) tumorigenesis was investigated. Immunohistochemistry and mutation analysis were performed using 18 cases of human primary OMM. Results revealed 16 cases positive to c-kit protein. Atypical melanocytes expressed c-kit. All in situ components expressed c-kit, but only four cases exhibited intense expression in the invasive component. Missense mutations were observed in four cases, and two of those correlated with increased protein expression. C-kit expression in atypical melanocytes suggests the role of c-kit in the early stage of OMM tumorigenesis. C-kit protein expression correlated with activating mutations indicating the pertinent role of the proto-oncogene KIT in the tumorigenesis of OMM.
Asunto(s)
Melanoma/metabolismo , Mucosa Bucal/metabolismo , Neoplasias de la Boca/metabolismo , Mutación Missense , Proteínas Proto-Oncogénicas c-kit/metabolismo , Secuencia de Bases , Análisis Mutacional de ADN , ADN de Neoplasias/análisis , Humanos , Inmunohistoquímica , Melanocitos/metabolismo , Melanocitos/patología , Melanoma/genética , Melanoma/patología , Datos de Secuencia Molecular , Mucosa Bucal/patología , Neoplasias de la Boca/genética , Neoplasias de la Boca/patología , Polimorfismo Conformacional Retorcido-Simple , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-kit/genética , Análisis de Secuencia de ADNRESUMEN
Head and neck squamous cell carcinoma (HNSCC) is a frequently occurring cancer, and despite improvement of its treatment methods, including chemotherapy, radiotherapy, and surgery, the improvement of survival remains poor. Recent advances in molecular biology of human cancer indicated various molecular abnormalities in HNSCC, including activation of oncogenes and inactivation of tumor suppressor genes (TSGs). Dickkopf (Dkk)-3 gene is known as a negative regulator of Wnt signaling and is suggested to function as TSG in several kinds of malignancies. We hypothesized that Dkk-3 might play an important role in HNSCC, too. Thus, in the current study, we analyzed allelic alteration of Dkk-3 locus (chromosome 11p15.2) by means of loss of heterozygosity (LOH) analysis. The study population consisted of 50 patients with HNSCC (mean age of 65 years old). Furthermore, we also examined the correlation between LOH findings of Dkk-3 locus with clinicopathological parameters to investigate its use as a biomarker in HNSCC. A remarkable LOH ratio (57%) was detected in the cases studied, implying that Dkk-3 is likely to be involved in HNSCC carcinogenesis. However, interestingly and in contrast to the expectations, we found that the group with LOH of Dkk-3 locus had less lymph node metastasis, and showed a favorable overall survival compared to the patients with retention of Dkk-3 area in survival analysis. These results indicate that Dkk-3 can play a role in HNSCC carcinogenesis with unknown mechanism. Moreover, allelic loss at Dkk-3 locus may also be used as a novel prognostic biomarker in HNSCC.