RESUMEN
BACKGROUND: Unicystic ameloblastoma, an odontogenic neoplasm, presents clinical and radiographic similarities with dentigerous and radicular cysts, non-neoplastic lesions. It is not always possible to reach a final diagnosis with the incisional biopsy, leading to inappropriate treatment. The BRAFV600E activating mutation has been reported in a high proportion of ameloblastomas. The purpose of the study was to assess the utility of the detection of the BRAFV600E mutation in the differential diagnosis of unicystic ameloblastoma with dentigerous and radicular cysts. METHODS: Twenty-six archival samples were included, comprising eight unicystic ameloblastomas (UAs), nine dentigerous and nine radicular cysts. The mutation was assessed in all samples by anti-BRAFV600E (clone VE1) immunohistochemistry (IHC) and by TaqMan mutation detection qPCR assay. Sanger sequencing was further carried out when samples showed conflicting results in the IHC and qPCR. RESULTS: Although all UAs (8/8) showed positive uniform BRAFV600E staining along the epithelial lining length, the mutation was not confirmed by qPCR and Sanger sequencing in three samples. Positive staining for the BRAFV600E protein was observed in one dentigerous cyst, but it was not confirmed by the molecular methods. Furthermore, 2/9 dentigerous cysts and 2/9 radicular cysts showed non-specific immunostaining of the epithelium or plasma cells. None of the dentigerous or radicular cysts cases presented the BRAFV600E mutation in the qPCR assay. CONCLUSIONS: The BRAFV600E antibody (clone VE1) IHC may show non-specific staining, but molecular assays may be useful for the diagnosis of unicystic ameloblastoma, in conjunction with clinical, radiological and histopathological features.
Asunto(s)
Ameloblastoma/diagnóstico , Ameloblastoma/genética , Neoplasias Maxilomandibulares/diagnóstico , Mutación , Proteínas Proto-Oncogénicas B-raf/genética , Quiste Radicular/diagnóstico , Adolescente , Adulto , Ameloblastoma/enzimología , Ameloblastoma/patología , Secuencia de Bases , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Humanos , Inmunohistoquímica , Neoplasias Maxilomandibulares/enzimología , Neoplasias Maxilomandibulares/genética , Neoplasias Maxilomandibulares/patología , Masculino , Persona de Mediana Edad , Tumores Odontogénicos/diagnóstico , Tumores Odontogénicos/enzimología , Tumores Odontogénicos/genética , Tumores Odontogénicos/patología , Quiste Radicular/enzimología , Quiste Radicular/genética , Quiste Radicular/patología , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto JovenRESUMEN
OBJECTIVE: To compare the proliferation and the expression of matrix metalloproteinases (MMPs; MMP-2 and MMP-9) in solid and unicystic ameloblastomas with ameloblastic carcinomas. STUDY DESIGN: Five cases of ameloblastic carcinoma (AC), 18 cases of solid ameloblastoma (SA), and seven of unicystic ameloblastoma (UA) were selected. The immunohistochemical expression of MMPs was assessed by the percentage of positive tumor cells and stained stroma. The mean argyrophilic nucleolar organizer region (AgNOR) and the percentage of cells with more than one AgNOR per nucleus were evaluated. RESULTS: Statistically significant higher mean AgNOR was observed in AC than in SA and UA. MMP-2 was expressed similarly in tumor and stroma among groups. MMP-9 was higher in the stroma of SA than that of UA (P = .0484). CONCLUSIONS: The cell proliferation was related to the greater aggressiveness of AC. High expression of MMP-2 and MMP-9 in all lesions highlighted the importance of these enzymes in the biology of ameloblastic tumors.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Mandibulares/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Ameloblastoma/patología , Biomarcadores de Tumor/metabolismo , Brasil , Proliferación Celular , Niño , Estudios Transversales , Femenino , Humanos , Masculino , Neoplasias Mandibulares/patología , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
Ameloblastoma is the most common benign odontogenic tumor in Japan. It is believed that it expands in the jaw bone through peritumoral activation of osteoclasts by receptor activator of nuclear factor kappa-B ligand (RANKL) released from the ameloblastoma, as in bone metastases of cancer cells. However, the clinical features of ameloblastoma, including its growth rate and patterns of invasion, are quite different from those of bone metastasis of cancer cells, suggesting that different underlying mechanisms are involved. Therefore, in the present study, we examined the possible mechanisms underlying the invasive expansion of ameloblastoma in the jaw bone. Expression levels of RANKL assessed by western blotting were markedly lower in ameloblastoma (AM-1) cells than in highly metastatic oral squamous cell carcinoma (HSC-3) cells. Experiments coculturing mouse macrophages (RAW264.7) with AM-1 demonstrated low osteoclastogenic activity, as assessed by tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cell formation, probably because of low release of RANKL, whereas cocultures of RAW264.7 with HSC-3 cells exhibited very high osteoclastogenic activity. Thus, RANKL release from AM-1 appeared to be too low to generate osteoclasts. However, AM-1 cultured directly on calcium phosphate-coated plates formed resorption pits, and this was inhibited by application of bafilomycin A1. Furthermore, vacuolar-type H+-ATPase (V-ATPase) and H+/Cl- exchange transporter 7 (CLC-7) were detected on the surface of AM-1 cells by plasma membrane biotinylation and immunofluorescence analysis. Immunohistochemical analysis of clinical samples of ameloblastoma also showed plasma membrane-localized V-ATPase and CLC-7 in the epithelium of plexiform, follicular and basal cell types. The demineralization activity of AM-1 was only 1.7% of osteoclasts demineralization activity, and the growth rate was 20% of human normal skin keratinocytes and HSC-3 cells. These results suggest that the slow expansion of several typical types of ameloblastomas in jaw bone is attributable to its slow growth and low demineralization ability.
Asunto(s)
Ameloblastoma/enzimología , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Neoplasias Maxilomandibulares/enzimología , Maxilares/enzimología , ATPasas de Translocación de Protón Vacuolares/metabolismo , Fosfatasa Ácida/metabolismo , Ameloblastoma/patología , Animales , Línea Celular Tumoral , Membrana Celular/enzimología , Humanos , Isoenzimas/metabolismo , Maxilares/patología , Neoplasias Maxilomandibulares/patología , Queratinocitos/citología , Ratones , Neoplasias de la Boca/enzimología , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Invasividad Neoplásica , Metástasis de la Neoplasia , Osteoclastos/metabolismo , Ligando RANK/metabolismo , Células RAW 264.7 , Fosfatasa Ácida TartratorresistenteRESUMEN
Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) have been implicated in the local invasiveness of ameloblastoma. This study aims to assess the role of MMP-9 and TIMP-2 in regulating tumor progression in ameloblastomas, taking tooth germs as control. Formalin-fixed, paraffin-embedded tissue sections of 4 tooth germs and 32 ameloblastomas were immunohistochemically examined using antibodies against MMP-9 and TIMP-2. Strong MMP-9 positivity was seen in the epithelial component in both controls and solid multicystic ameloblastoma. Statistically significant difference was observed in the mean stromal MMP-9 immunoscores between follicular, acanthomatous, and granular ameloblastoma when compared with the tooth germ (P=0.004). TIMP-2 expression in the epithelial and mesenchymal components of solid multicystic ameloblastoma and tooth germ was weak as compared with MMP-9 expression. Highest mean epithelial TIMP-2 immunoscore was observed in follicular ameloblastoma and the difference was statistically significant between follicular and granular ameloblastoma (P=0.05). The comparison of mean stromal TIMP-2 immunoscores showed statistically significant difference between follicular subtype and tooth germ (P=0.048), with tooth germ showing least expression among the groups studied. Strong stromal expression of MMP-9 in ameloblastoma compared with tooth germ mesenchyme indicated the possibility of tumor induction with release of growth factors and cytokines, resulting in invasiveness of ameloblastoma. Epithelial TIMP-2 expression was associated with the least and most aggressive behavior of follicular and granular cell ameloblastoma, respectively. Stromal TIMP-2 expression reflected its role in regulating tumor progression in ameloblastoma and in regulating developmental processes in tooth germs by their inhibitory effect on MMP-9.
Asunto(s)
Ameloblastoma/patología , Metaloproteinasa 9 de la Matriz/metabolismo , Odontogénesis , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Ameloblastoma/enzimología , Ameloblastoma/metabolismo , Humanos , Inmunohistoquímica , MorfogénesisRESUMEN
BACKGROUND: Ameloblastoma (AM) is a benign odontogenic neoplasm characterized by local invasiveness and recurrence. We compared the immunohistochemical expression of matrix metalloproteinases in different clinical types of AM as well as in normal odontogenic tissue. METHODS: Thirteen cases of solid AMs, five cases of unicystic AM and eight pericoronal follicles (PF) were selected and subjected to immunohistochemical investigation for matrix metalloproteinase-1, matrix metalloproteinase-2 and matrix metalloproteinase-9 expressions. RESULTS: The expressions of MMP-1 and MMP-2 were very high in the cytoplasm of cells throughout the entire epithelium and in fibroblasts from the adjacent connective tissue. MMP-9 expression was observed in the same location although with weaker staining. The Kruskal-Wallis test showed statistically significant differences in the epithelial expressions of MMP-1 and MMP-2; there was lower expression among solid AMs when compared with unicystic AM and PF. Compared to both types of AM, higher stromal expression of MMP-9 was found in PF. CONCLUSION: MMP-1, MMP-2 and MMP-9 seem to be associated with AM tumour behaviour as well as physiological tissue remodelling within PF.
Asunto(s)
Ameloblastoma/enzimología , Saco Dental/enzimología , Neoplasias Maxilomandibulares/enzimología , Metaloproteinasas de la Matriz/biosíntesis , Tumores Odontogénicos/enzimología , Ameloblastoma/metabolismo , Ameloblastoma/patología , Tejido Conectivo/enzimología , Tejido Conectivo/patología , Saco Dental/metabolismo , Saco Dental/patología , Epitelio/enzimología , Epitelio/metabolismo , Epitelio/patología , Fibroblastos/enzimología , Fibroblastos/metabolismo , Fibroblastos/patología , Humanos , Inmunohistoquímica , Neoplasias Maxilomandibulares/metabolismo , Neoplasias Maxilomandibulares/patología , Metaloproteinasa 1 de la Matriz/biosíntesis , Metaloproteinasa 2 de la Matriz/biosíntesis , Metaloproteinasa 9 de la Matriz/biosíntesis , Recurrencia Local de Neoplasia/enzimología , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Tumores Odontogénicos/metabolismo , Tumores Odontogénicos/patologíaRESUMEN
BACKGROUND: Odontogenic tumours are a heterogeneous group of lesions formed from tissues that give rise to the tooth. DNA methylation, a covalent addition of a methyl group to the 5-carbon position of a cytosine nucleotide, is considered an important regulator of gene expression. The addition of the methyl radical is catalysed by DNA methyltransferases (DNMTs). Although some epigenetic studies have been conducted in odontogenic tumours, a study with the three types of DNMTs in several different members of this group is missing. This study analyses the expression of DNMTs in odontogenic tumours. METHODS: Formalin-fixed and paraffin-embedded tissue samples of 20 ameloblastomas, 10 calcifying cystic odontogenic tumours, 10 calcifying epithelial tumours, 10 adenomatoid odontogenic tumours, 10 keratocystic odontogenic tumours, five ameloblastic fibromas, two ameloblastic fibro-odontomas, four central odontogenic fibromas, seven peripheral odontogenic fibromas and 10 odontogenic myxomas were included. Immunohistochemical expression of DNMT1, 3A and 3B was assessed using a semi-quantitative analysis, and also a correlation with p21, p27 and E-cadherin immunoexpression was made. RESULTS: DNMT1, 3A and 3B were expressed in the nucleus and/or cytoplasm of all odontogenic tumours. DNMT1 expression was directly correlated with p27 expression in ameloblastomas. CONCLUSION: The high expression of DNMTs in odontogenic tumour cells suggests methylation as an important mechanism for this group of tumours.
Asunto(s)
ADN (Citosina-5-)-Metiltransferasas/análisis , Tumores Odontogénicos/enzimología , Adolescente , Adulto , Anciano , Ameloblastoma/química , Ameloblastoma/enzimología , Cadherinas/análisis , Núcleo Celular/química , Núcleo Celular/enzimología , Niño , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/análisis , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/análisis , Citoplasma/química , Citoplasma/enzimología , ADN (Citosina-5-)-Metiltransferasa 1 , ADN Metiltransferasa 3A , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Tumores Odontogénicos/química , Adulto Joven , ADN Metiltransferasa 3BRESUMEN
Ameloblastoma is a benign but locally infiltrative odontogenic neoplasm. Although ameloblastomas rarely metastasise, recurrences together with radical surgery often result in facial deformity and significant morbidity. Development of non-invasive therapies has been precluded by a lack of understanding of the molecular background of ameloblastoma pathogenesis. When addressing the role of ERBB receptors as potential new targets for ameloblastoma, we discovered significant EGFR over-expression in clinical samples using real-time RT-PCR, but observed variable sensitivity of novel primary ameloblastoma cells to EGFR-targeted drugs in vitro. In the quest for mutations downstream of EGFR that could explain this apparent discrepancy, Sanger sequencing revealed an oncogenic BRAF V600E mutation in the cell line resistant to EGFR inhibition. Further analysis of the clinical samples by Sanger sequencing and BRAF V600E-specific immunohistochemistry demonstrated a high frequency of BRAF V600E mutations (15 of 24 samples, 63%). These data provide novel insight into the poorly understood molecular pathogenesis of ameloblastoma and offer a rationale to test drugs targeting EGFR or mutant BRAF as novel therapies for ameloblastoma.
Asunto(s)
Ameloblastoma/genética , Biomarcadores de Tumor/genética , Neoplasias Maxilomandibulares/genética , Mutación , Proteínas Proto-Oncogénicas B-raf/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Ameloblastoma/tratamiento farmacológico , Ameloblastoma/enzimología , Ameloblastoma/patología , Antineoplásicos/farmacología , Biomarcadores de Tumor/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Resistencia a Antineoplásicos , Receptores ErbB/antagonistas & inhibidores , Receptores ErbB/genética , Receptores ErbB/metabolismo , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Neoplasias Maxilomandibulares/tratamiento farmacológico , Neoplasias Maxilomandibulares/enzimología , Neoplasias Maxilomandibulares/patología , Masculino , Persona de Mediana Edad , Terapia Molecular Dirigida , Selección de Paciente , Fenotipo , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas B-raf/metabolismo , Receptor ErbB-4 , Transducción de Señal/efectos de los fármacos , Adulto JovenRESUMEN
Rho GTPases are proteins that regulate cell cycle, shape, polarization, invasion, migration, and apoptosis, which are important characteristics of normal and neoplastic cells. Rho GTPases expression has been reported in normal tooth germ and several pathologies; however, it has not been evaluated in ameloblastomas. The aim of this study was to analyze the expression and distribution of RhoA, RhoB, Rac1, and Cdc42 Rho GTPases in solid and unicystic ameloblastomas. Three-micrometer sections from paraffin-embedded specimens were evaluated by using an avidin-biotin immunohistochemical method with antibodies against the proteins mentioned above. RhoA and RhoB staining was observed in a high number of cells (P < 0.05) and greater intensity in non-polarized ones. Rac1 was not observed, and Cdc42 did not show any statistical differences between the number of non-polarized and basal positive cells (P > 0.05). Upon comparing the studied ameloblastomas, a higher number of positive cells in the unicystic variant was observed than that in the solid one (P < 0,05). The results obtained suggest that these GTPases could play a role in the ameloblastoma neoplastic epithelial cell phenotype determination (polarized or non-polarized), as well as in variant (solid or unicystic) and subtype (follicular or plexiform) determination. Furthermore, they could participate in solid ameloblastoma invasion mechanisms.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Maxilomandibulares/enzimología , Proteínas de Unión al GTP rho/metabolismo , Ameloblastoma/patología , Humanos , Inmunohistoquímica , Neoplasias Maxilomandibulares/patología , Distribución Tisular , Proteína de Unión al GTP cdc42/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Proteína de Unión al GTP rhoA/metabolismo , Proteína de Unión al GTP rhoB/metabolismoRESUMEN
OBJECTIVE: The aim of this study was to evaluate the immunohistochemical expression of collagen IV, matrix metalloproteinase (MMP) 9 and tissue inhibitor of MMP (TIMP) 2 in dentigerous cysts (DCs), radicular cysts (RCs), keratocystic odontogenic tumors (KOTs), and ameloblastomas. STUDY DESIGN: Twenty cases of DCs, 20 RCs, 20 KOTs, and 20 ameloblastomas were selected and analyzed by immunohistochemistry. RESULTS: Most DCs and RCs showed continuous and >50% staining for collagen IV in the basement membrane of the epithelium, whereas predominantly discontinuous thin and ≤ 50% staining was observed in KOTs and ameloblastomas, with a significant difference in staining percentage (P < .001). MMP-9 was diffusely distributed and localized in both epithelial and mesenchymal cells of all of the lesions analyzed. The staining percentage was higher in the epithelium (P = .058) and mesenchyme (P = .005) of KOTs and ameloblastomas. Moreover, the distribution pattern, location, and percentage of expression of TIMP-2 were similar in the lesions studied, except for ameloblastoma, with a significant difference in staining percentage (P < .001). CONCLUSION: These results demonstrate that the interaction between collagen IV, MMP-9, and TIMP-2 is an important factor for the establishment of differences in the biologic behavior of the odontogenic cysts and tumors studied.
Asunto(s)
Ameloblastoma/patología , Colágeno Tipo IV/análisis , Metaloproteinasa 9 de la Matriz/análisis , Quistes Odontogénicos/patología , Inhibidores de Proteasas/análisis , Inhibidor Tisular de Metaloproteinasa-2/análisis , Ameloblastoma/enzimología , Membrana Basal/enzimología , Membrana Basal/patología , Colorantes , Tejido Conectivo/enzimología , Tejido Conectivo/patología , Quiste Dentígero/enzimología , Quiste Dentígero/patología , Células Endoteliales/enzimología , Células Endoteliales/patología , Células Epiteliales/enzimología , Células Epiteliales/patología , Epitelio/enzimología , Epitelio/patología , Fibroblastos/enzimología , Fibroblastos/patología , Humanos , Inmunohistoquímica , Mesodermo/enzimología , Mesodermo/patología , Quistes Odontogénicos/enzimología , Quiste Radicular/enzimología , Quiste Radicular/patologíaRESUMEN
Heparanase is an endoglycosidase that cleaves heparan sulfate (HS), thus participating in degradation and remodeling of the extracellular matrix (ECM). Heparanase up-regulation is correlated with lymph node and distant metastasis, microvessel density and reduced postoperative survival of cancer patients. In the present study, we carried out an immunohistochemical investigation of heparanase to extend and confirm present knowledge regarding its expression in ameloblastomas (AMs), which are characterized by locally aggressive behavior. Paraffin-embedded tissue specimens of 53 AMs were stained using an antibody against heparanase. Immunohistochemical reactivity for heparanase was detected in 94.3% of the AMs examined. Heparanase was expressed strongly in peripheral columnar cells and slightly in central stellate reticulum-like cells. Small tumor nests and budding epithelial branches showed a stronger staining pattern. Stromal cells were negative for heparanase, or showed diffuse expression. However, an enhanced positive immunoreaction was present specifically near osseous tissue and adjacent to the invasive front of tumor nests. Areas of cystic degeneration showed intense heparanase immunoreactivity. The enzyme may facilitate the function of HS-binding growth factors that elicit an angiogenic response and favor osteoclastogenesis in AM.
Asunto(s)
Ameloblastoma/enzimología , Glucuronidasa/biosíntesis , Neoplasias Maxilomandibulares/enzimología , Adolescente , Adulto , Anciano , Ameloblastoma/patología , Niño , Matriz Extracelular/metabolismo , Femenino , Humanos , Técnicas para Inmunoenzimas , Neoplasias Maxilomandibulares/patología , Masculino , Persona de Mediana Edad , Invasividad Neoplásica/patología , Neovascularización Patológica , Osteoclastos/enzimología , Adhesión en Parafina , Regulación hacia Arriba , Adulto JovenRESUMEN
BACKGROUND: Ameloblastoma is the most common clinically-significant epithelial odontogenic tumor, and is considered a benign but locally-aggressive tumor of the craniofacial region. Osteonectin/secreted protein acidic and rich in cysteine (SPARC) is induced in response to a number of biological processes such as tumor growth and metastasis, whereas matrix metalloproteinases (MMPs) degrade the extracellular matrix and participate in various biological processes including tumor invasion and metastasis. We hypothesize that SPARC acts with MMPs for the local invasiveness of ameloblastoma. The aim of this study was to examine the association of SPARC with MMP-1, MMP-2, and MMP-9 in ameloblastoma. METHOD: Immunohistochemical expression of SPARC, MMP-1, MMP-2, and MMP-9 as well as co-expression of SPARC and MMP-9 were examined in a cohort of 23 cases of ameloblastoma. RESULTS: SPARC, MMP-1, -2, and -9 were detected in the cytoplasm of the ameloblastic-like columnar cells and stellate-reticulum-like cells as well as in the stromal tissues of fibroblasts and endothelial cells of our cohort of ameloblastoma patients. Furthermore, co-expression of SPARC and MMP-9 were found in 23 cases of ameloblastoma. This may be the first study to demonstrate that the expression level of SPARC was statistically correlated with MMP-9 but not with MMP-1 or -2 in ameloblastoma. CONCLUSION: Our results suggest a putative association between SPARC and MMPs (especially MMP-9) in ameloblastoma to regulate tumor invasion.
Asunto(s)
Ameloblastoma/patología , Metaloproteinasas de la Matriz Secretadas/análisis , Osteonectina/análisis , Adolescente , Adulto , Ameloblastoma/enzimología , Estudios de Cohortes , Células Endoteliales/patología , Femenino , Fibroblastos/patología , Estudios de Seguimiento , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Masculino , Metaloproteinasa 1 de la Matriz/análisis , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/análisis , Persona de Mediana Edad , Invasividad Neoplásica , Recurrencia Local de Neoplasia/patología , Adulto JovenRESUMEN
Ameloblastoma (AM) is recognized as a benign tumour but locally invasive with a high risk of recurrence. In vitro model systems for studying AM are limited due to the fact that AM cells grow poorly and begin to senesce early. Japanese researchers have reported the construction of an AM cell line, AM-1, by exposing cells to human papillomavirus 16 (HPV16) but retaining the potential of transformation. In this study, we used a retroviral infection method to over-express the human telomerase reverse transcriptase (hTERT) gene to acquire immortality of hTERT(+)-AM cells. Furthermore, it was revealed both by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot that the pathway of immortalization was loss of p16, not p53 or p21. Also, there was no evidence indicating that the hTERT(+)-AM cells underwent malignant transformation by the nude mouse tumorigenicity assay. Taken together, this hTERT-immortalized cell line may be a potentially valuable and reliable cell model for further study of the invasive properties of AM in vitro.
Asunto(s)
Ameloblastoma/patología , Neoplasias Maxilomandibulares/patología , Proteínas de Neoplasias/metabolismo , Telomerasa/metabolismo , Ameloblastoma/enzimología , Ameloblastoma/genética , Western Blotting , Técnicas de Cultivo de Célula , Línea Celular Transformada/virología , Línea Celular Tumoral , Proliferación Celular , Transformación Celular Viral , Activación Enzimática , Epigénesis Genética , Papillomavirus Humano 16 , Humanos , Neoplasias Maxilomandibulares/enzimología , Neoplasias Maxilomandibulares/genética , Masculino , Persona de Mediana Edad , Proteínas de Neoplasias/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Telomerasa/genética , Transfección/métodosRESUMEN
OBJECTIVE: The aim was to evaluate the expression of matrix metalloproteinases (MMPs) 7 and 26 in ameloblastomas and adenomatoid odontogenic tumors (AOTs). STUDY DESIGN: Twenty intraosseous solid ameloblastomas and 10 intraosseous AOTs were evaluated regarding immunohistochemical expression of MMP-7 and -26 in the epithelium and stroma. RESULTS: There was no statistically significant difference in MMP-7 and -26 expression between the epithelium of ameloblastomas (P = .50) and AOTs (P = .90). Stromal staining for MMP-7 was evident in all cases. For MMP-26, stromal staining was observed in 65% of ameloblastomas and 50% of AOTs, and this difference was not statistically significant (P = .69). CONCLUSION: The marked expression of these matrilysins suggests their role in the process of tissue remodeling and growth in the studied tumors, but it does not relate to the their distinct patterns of aggressiveness.
Asunto(s)
Ameloblastoma/enzimología , Metaloproteinasa 7 de la Matriz/análisis , Metaloproteinasas de la Matriz Secretadas/análisis , Tumores Odontogénicos/enzimología , Tejido Conectivo/enzimología , Células Endoteliales/enzimología , Células Epiteliales/enzimología , Epitelio/enzimología , Fibroblastos/enzimología , Humanos , InmunohistoquímicaRESUMEN
BACKGROUND: Ameloblastoma is an odontogenic benign tumor characterized by local invasiveness and most of its local recurrences clinically result from local invasion. This study used matrix metalloproteinase-2 (MMP-2) inhibitor I (MMP-2I) to investigate the role played by MMP-2 activity in the local invasiveness of ameloblastoma. METHODS: The cells and xenografts of ameloblastoma were treated with MMP-2I and treatment group were compared with the control group. In vitro, the invasive activity of tumor cells was assayed in transwell cell culture chamber. Gelatinolytic activity of gelatinases and MMP-2/tissue inhibitor of matrix metalloproteinase (TIMP-2) protein expression was detected using gelatin zymography and flow cytometry. The cell viability and adhesion were evaluated using methyl thiazol tetrazolium. In vivo, bilateral subrenal capsule xenograft transplantation of ameloblastoma was performed in 10 nude mice and the invasion of ameloblastoma into the renal parenchyma was observed. RESULTS: Active-MMP-2 of conditioned media was significantly lower in treatment group than in the control group. Accordingly, potential of in vitro cell invasion, adhesion and in vivo tumor invasion were also significantly lower in the treatment group than in the control group. CONCLUSIONS: Inhibitor of MMP-2 activity suppressed the local invasive capability of ameloblastoma by decreasing MMP-2 activity. MMP-2 activity is in relation with invasive capacity of ameloblastoma.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Maxilomandibulares/enzimología , Inhibidores de la Metaloproteinasa de la Matriz , Inhibidor Tisular de Metaloproteinasa-2/farmacología , Ameloblastoma/tratamiento farmacológico , Animales , Humanos , Neoplasias Maxilomandibulares/tratamiento farmacológico , Metaloproteinasa 2 de la Matriz/fisiología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Invasividad Neoplásica , Trasplante de Neoplasias , Inhibidor Tisular de Metaloproteinasa-2/uso terapéutico , Células Tumorales CultivadasRESUMEN
Myofibroblasts are frequent in the stroma of neoplasm and by the expression of proteinases they can influence tumor infiltration and progression. In the present study, presence of myofibroblasts and expression of matrix metalloproteinase-2 (MMP-2) and urokinase plasminogen activator (uPA) were examined in intra-osseous solid multicystic ameloblastomas to determine their roles in the clinicopathological features of the tumors. Fifty seven ameloblastomas were analyzed immunohistochemically with antibodies against the isoform alpha of the smooth muscle actin (alpha-SMA), a specific marker of myofibroblasts, MMP-2 and uPA. Myofibroblasts were found in the stroma, in close contact with neoplastic cell islands, of approximately 58% (n = 33) of the ameloblastomas. MMP-2 and uPA were found in the cytoplasm of both neoplastic and stromal cells. A significant correlation between presence of myofibroblasts and MMP-2 expression was observed. Abundant presence of myofibroblast in the stroma of the tumors and expression of MMP-2 in the neoplastic or stromal cells were significantly correlated with rupture of the osseous cortical, which has been considered an important prognostic marker of ameloblastoma aggressiveness. Ours results suggest that abundant presence of myofibroblasts and expression of MMP-2 in solid ameloblastomas may be associated with a more aggressive infiltrative behavior.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Óseas/enzimología , Fibroblastos/patología , Metaloproteinasa 2 de la Matriz/metabolismo , Adolescente , Adulto , Anciano , Ameloblastoma/patología , Biomarcadores de Tumor/metabolismo , Neoplasias Óseas/patología , Niño , Citoplasma/metabolismo , Progresión de la Enfermedad , Femenino , Fibroblastos/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Músculo Liso/enzimología , Pronóstico , Células del Estroma/enzimología , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Ameloblastomas are odontogenic neoplasms characterized by local invasiveness. This study was conducted to address the role of matrix metalloproteinase-2 (MMP-2) in the invasiveness of ameloblastomas. METHODS: Plasmids containing either MMP-2 siRNA or tissue inhibitor of metalloproteinase-2 (TIMP-2) cDNA were created and subsequently transfected into primary ameloblastoma cells. Zymography, RT-PCR, and Western blots were used to assess MMP-2 activity and expression of MMP-2 and TIMP-2, as well as protein levels. RESULTS: Primary cultures of ameloblastoma cells expressed cytokeratin (CK) 14 and 16, and MMP-2, but only weakly expressed CK18 and vimentin. MMP-2 mRNA and protein levels were significantly inhibited by RNA interference (P < 0.05). Both MMP-2 siRNA and TIMP-2 overexpression inhibited MMP-2 activity and the in vitro invasiveness of ameloblastoma. CONCLUSION: These results indicate that inhibition of MMP-2 activity suppresses the local invasiveness of ameloblastoma cells. This mechanism may serve as a novel therapeutic target in ameloblastomas pursuant to additional research.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Maxilomandibulares/enzimología , Inhibidores de la Metaloproteinasa de la Matriz , Ameloblastoma/genética , Ameloblastoma/patología , Humanos , Neoplasias Maxilomandibulares/genética , Neoplasias Maxilomandibulares/patología , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/metabolismo , Invasividad Neoplásica , Plásmidos/genética , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Inhibidor Tisular de Metaloproteinasa-2/biosíntesis , Inhibidor Tisular de Metaloproteinasa-2/genética , Inhibidor Tisular de Metaloproteinasa-2/metabolismo , Transfección , Células Tumorales CultivadasRESUMEN
BACKGROUND: To evaluate roles of mitogen-activated protein kinases (MAPKs) in oncogenesis and cytodifferentiation of odontogenic tumors, expression of phosphorylated JNK (p-JNK), p38 MAPK (p-p38 MAPK), and ERK5 (p-ERK5) was analyzed in ameloblastic tumors as well as in tooth germs. METHODS: Ten tooth germs, 47 ameloblastomas, and 5 malignant ameloblastic tumors were examined immunohistochemically with the antibodies against p-JNK, p-p38 MAPK, and p-ERK5. RESULTS: Immunoreactivity for p-JNK was detected in epithelial or neoplastic cells detached from the basement membrane in 7 tooth germs and 7 ameloblastomas, and the expression levels of p-JNK in ameloblastic tumors were significantly lower than that in tooth germs. Expression of p-p38 MAPK was found in epithelial or neoplastic cells in tooth germs and ameloblastic tumors except for two ameloblastomas, and increased expression was found in keratinizing cells of acanthomatous ameloblastomas. The expression level of p-p38 MAPK in ameloblastomas was significantly higher than the levels in tooth germs and malignant ameloblastic tumors. Immunoreactivity for p-ERK5 was found predominantly in epithelial or neoplastic cells near the basement membrane in tooth germs and ameloblastic tumors. The expression levels of p-ERK5 in ameloblastic tumors were slightly higher than that in tooth germs, and plexiform ameloblastomas showed significantly higher p-ERK5 expression than follicular ameloblastomas. CONCLUSION: Expression of p-JNK, p-p38 MAPK, and p-ERK5 in tooth germs and ameloblastic tumors suggests that these MAPK signaling pathways contribute to cell proliferation, differentiation, or apoptosis in both normal and neoplastic odontogenic tissues. Altered expression of these phosphorylated MAPKs in ameloblastic tumors may be involved in oncogenesis and tumor cell differentiation.
Asunto(s)
Ameloblastoma/enzimología , Neoplasias Maxilomandibulares/enzimología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Germen Dentario/enzimología , Diferenciación Celular , Proliferación Celular , Humanos , Inmunohistoquímica , Proteínas Quinasas JNK Activadas por Mitógenos/análisis , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Proteína Quinasa 7 Activada por Mitógenos/análisis , Proteína Quinasa 7 Activada por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/análisis , Fosforilación , Proteínas Quinasas p38 Activadas por Mitógenos/análisis , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
BACKGROUND: To evaluate the roles of angiogenic factors in the development and progression of odontogenic tumors, expression of platelet-derived endothelial cell growth factor/thymidine phosphorylase (PD-ECGF/TP) and of angiopoietins in ameloblastic tumors as well as in tooth germs. METHODS: Tissue specimens of 11 tooth germs, 44 ameloblastomas, and five malignant ameloblastic tumors were examined immunohistochemically with the use of antibodies against PD-ECGF/TP and angiopoietin-1 and -2. RESULTS: Immunohistochemical reactivity for PD-ECGF/TP was detected in mesenchymal cells in tooth germs and stromal cells in ameloblastic tumors, and the level of immunoreactivity for PD-ECGF/TP was significantly higher in ameloblastomas than in tooth germs. Granular cell ameloblastomas showed PD-ECGF/TP reactivity in granular neoplastic cells as well as in stromal cells. Immunoreactivity for angiopoietin-1 and -2 was detected predominantly in odontogenic epithelial cells near the basement membrane in tooth germs and in benign and malignant ameloblastic tumors. Malignant ameloblastic tumors had decreased angiopoietin-1 reactivity and ameloblastic carcinomas had increased angiopoietin-2 reactivity as compared with the respective levels in tooth germs and ameloblastomas. Immunohistochemical reactivity for angiopoietin-2 was slightly higher in follicular ameloblastomas than in plexiform ameloblastomas. CONCLUSION: Expression of PD-ECGF/TP and angiopoietin-1 and -2 in tooth germs and ameloblastic tumors suggests that these angiogenic factors participate in tooth development and odontogenic tumor progression by regulating angiogenesis. Altered expression of PD-ECGF/TP and angiopoietins in ameloblastic tumors may be involved in oncogenesis, malignant potential, and tumor cell differentiation.
Asunto(s)
Ameloblastoma/química , Angiopoyetina 1/análisis , Angiopoyetina 2/análisis , Neoplasias Maxilomandibulares/química , Timidina Fosforilasa/análisis , Ameloblastoma/enzimología , Humanos , Neoplasias Maxilomandibulares/enzimología , Células del Estroma/química , Células del Estroma/enzimología , Germen Dentario/química , Germen Dentario/enzimologíaRESUMEN
BACKGROUND: To evaluate the roles of matrix-degrading proteinase regulators in progression of odontogenic tumors, expression of membrane-bound matrix metalloproteinase (MMP) MT1-MMP, MMP inhibitor RECK and MMP inducer EMMPRIN was analyzed in ameloblastic tumors as well as in tooth germs. METHODS: Tissue specimens of 11 tooth germs, 40 ameloblastomas, and five malignant ameloblastic tumors were examined immunohistochemically with the use of antibodies against MT1-MMP, RECK, and EMMPRIN. RESULTS: Immunohistochemical reactivity for MT1-MMP, RECK and EMMPRIN was detected predominantly in odontogenic epithelial cells near the basement membrane in tooth germs and benign and malignant ameloblastic tumors. The level of immunoreactivity for MT1-MMP was slightly higher in benign and malignant ameloblastic tumors than in tooth germs. RECK expression was lower in ameloblastomas than in tooth germs. Follicular ameloblastomas showed significantly lower expression of RECK than plexiform ameloblastomas, and immunoreactivity for RECK in acanthomatous ameloblastomas was slightly lower than that in other cellular variants. CONCLUSION: Expression of MT1-MMP, RECK and EMMPRIN in tooth germs and ameloblastic tumors suggests that these normal and neoplastic epithelial components control MMP-dependent extracellular matrix (ECM) degradation during tooth development and tumor progression via epithelial-mesenchymal interactions.
Asunto(s)
Ameloblastoma/química , Basigina/análisis , Neoplasias Maxilomandibulares/química , Metaloproteinasas de la Matriz/análisis , Glicoproteínas de Membrana/análisis , Germen Dentario/química , Ameloblastoma/enzimología , Proteínas Ligadas a GPI , Humanos , Inmunohistoquímica , Neoplasias Maxilomandibulares/enzimología , Metaloproteinasas de la Matriz Asociadas a la Membrana , Germen Dentario/enzimologíaRESUMEN
Ameloblastoma is the most common odontogenic neoplasm, particularized by its local invasiveness. Heparanase is the endo-glucuronidase enzyme that specifically cleaves heparan sulfate, the important modulator of extracellular matrix, and related to invasion of tumor cells. In this study, we addressed to show the gene expression and localization of heparanase in ameloblastoma. Immunohistochemistry and in situ hybridization of heparanase were carried out in 23 ameloblastomas. Strong expression of heparanase at both mRNA and protein levels was detected in all ameloblastomas studied. Small tumor nests and budding epithelial branches showed stronger staining pattern and the stromal tissues at the immediate vicinity of the tumor nests with strong heparanase expression were loose and edematous. Cystic areas and squamous metaplastic areas of the tumor showed intense staining with heparanase antibody proposing the implication of heparanase in these processes. These results suggest the possible contribution of heparanase in the local invasiveness and secondary morphologic changes of ameloblastoma.