RESUMO
Calcifying odontogenic cyst (COC), once called calcifying cystic odontogenic tumor, is classified under the category of odontogenic cysts. However, the proliferative capacity of the lesional epithelium and consistent nuclear ß-catenin expression raise questions about its current classification. This study aimed to determine whether COC would be better classified as a neoplasm in the histologic and molecular context. Eleven odontogenic lesions diagnosed as COC or calcifying cystic odontogenic tumor were included in this study. The growth patterns of the lesional epithelium were analyzed histologically in all cases. ß-catenin immunohistochemistry and molecular profiling using Sanger sequencing and whole-exome sequencing were performed in 10 cases. Of the 11 cases studied, histologic features reminiscent of so-called adenoid ameloblastoma were observed in 72.7% (8/11), and small islands of clear cells extended into the wall in 36.4% (4/11). Intraluminal and/or mural epithelial proliferation was found in 72.7% of the cases (8/11). Nuclear ß-catenin expression was observed focally in all 10 cases studied, mainly highlighting epithelial cells forming morules and adjacent to dentinoid. CTNNB1 hotspot mutations were detected in 60.0% of the cases (6/10). All the remaining cases had frameshift mutations in tumor-suppressor genes involved in the WNT pathway, including APC and NEDD4L. Recurrent WNT pathway mutations leading to nuclear translocation of ß-catenin and distinct epithelial growth patterns found in COC are the neoplastic features shared by its solid counterpart, dentinogenic ghost cell tumor, supporting its classification as a tumor rather than a cyst.
Assuntos
Mutação , Cisto Odontogênico Calcificante , Via de Sinalização Wnt , Humanos , Feminino , Masculino , Cisto Odontogênico Calcificante/patologia , Cisto Odontogênico Calcificante/genética , Adulto , Via de Sinalização Wnt/genética , Pessoa de Meia-Idade , beta Catenina/genética , beta Catenina/metabolismo , Ameloblastoma/genética , Ameloblastoma/patologia , Ameloblastoma/metabolismo , Adolescente , Adulto Jovem , Neoplasias Maxilomandibulares/genética , Neoplasias Maxilomandibulares/patologia , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/análise , Tumores Odontogênicos/genética , Tumores Odontogênicos/patologia , Idoso , CriançaRESUMO
BACKGROUND: Ameloblastoma is an aggressively growing, highly recurrent odontogenic jaw tumor. Its association with BRAFV600E mutation is an indication for BRAFV00E-inhibitor therapy The study objective was to identify a sensitive low-cost test for BRAFV600E-positive ameloblastoma. We hypothesized that immunohistochemical staining of formalin-fixed paraffin-embedded tissues for BRAFV600E mutation is a low-cost surrogate for BRAFV600E gene sequencing when laboratory resources are inadequate for molecular testing. METHODS: Tissues from 40 ameloblastoma samples were retrieved from either formalin-fixed paraffin-embedded blocks, RNAlater™ stabilization solution or samples inadvertently pre-fixed in formalin before transfer to RNAlater™. BRAFV600E mutation was assessed by Direct Sanger sequencing, Amplification Refractory Mutation System-PCR and immunohistochemistry (IHC). RESULTS: BRAFV600E mutation was detected by IHC, Amplification Refractory Mutation System-PCR and Direct Sanger sequencing in 93.33%, 52.5% and 30% of samples respectively. Considering Direct Sanger sequencing as standard BRAFV600E detection method, there was significant difference between the three detection methods (ð2 (2) = 31.34, p < 0.0001). Sensitivity and specificity of IHC were 0.8 (95% CI: 0.64-0.90) and 0.9 (95% CI: 0.75-0.99) respectively, while positive predictive value and negative predictive value (NPV) were 0.9 and 0.8 (Fischer's test, p < 0.0001) respectively. Sensitivity and specificity of Amplification Refractory Mutation System-PCR detection method were 0.7 (95% CI: 0.53-0.80) and 0.9 (95% CI = 0.67-0.98) respectively, while PPV and NPV were 0.9 and 0.6 respectively (Fischer's test, p < 0.0001). CONCLUSION: Low-cost and less vulnerability of IHC to tissue quality make it a viable surrogate test for BRAFV600E detection in ameloblastoma. Sequential dual IHC and molecular testing for BRAFV600E will reduce equivocal results that could exclude some patients from BRAFV600E-inhibitor therapies.
Assuntos
Ameloblastoma , Tumores Odontogênicos , Humanos , Ameloblastoma/diagnóstico , Ameloblastoma/genética , Ameloblastoma/patologia , Proteínas Proto-Oncogênicas B-raf/genética , Mutação , Tumores Odontogênicos/genética , FormaldeídoRESUMO
BACKGROUND: Ameloblastoma and ameloblastic carcinoma are epithelial odontogenic tumors that can be morphologically similar. In the present study, we evaluated the DNA content and Ki-67 index in the two tumors. METHODS: The paraffin blocks of the tumors were selected to obtain sections for the immunohistochemical reactions and preparation of the cell suspension for acquisition in a flow cytometer. The Random Forest package of the R software was used to verify the contribution of each variable to classify lesions into ameloblastoma or ameloblastic carcinoma. RESULTS: Thirty-two ameloblastoma and five ameloblastic carcinoma were included in the study. In our sample, we did not find statistically significant differences in Ki-67 labeling rates. A higher fraction of cells in 2c (G1) was correlated with the diagnosis of ameloblastoma, whereas higher rates of 5c-exceeding rate (5cER) were correlated with ameloblastic carcinoma. The Random Forest model highlighted histopathological findings and parameters of DNA ploidy study as important features for distinguishing ameloblastoma from ameloblastic carcinoma. CONCLUSION: Our findings suggest that the parameters of the DNA ploidy study can be ancillary tools in the classification of ameloblastoma and ameloblastic carcinoma.
Assuntos
Ameloblastoma , Carcinoma , Tumores Odontogênicos , Humanos , Ameloblastoma/diagnóstico , Ameloblastoma/genética , Ameloblastoma/patologia , Antígeno Ki-67/genética , Tumores Odontogênicos/genética , Carcinoma/patologia , Ploidias , DNARESUMO
Ghost cell odontogenic carcinoma (GCOC) is defined as a rare type of odontogenic carcinoma that is characterized by ghost cells and occasional dentinoid. However, the current classification system based primarily on the presence of ghost cells has limitations in the diagnosis of GCOC and its histologic mimics including odontogenic carcinoma with dentinoid (OCD). This study reviewed previous studies on ß-catenin nuclear translocation and WNT pathway mutations in GCOC and OCD and discussed the potential utility of a new molecular-based classification "WNT pathway-altered malignant odontogenic tumor" for these rare odontogenic tumors.
Assuntos
Mutação , Tumores Odontogênicos , Via de Sinalização Wnt , beta Catenina , Humanos , beta Catenina/genética , beta Catenina/metabolismo , Tumores Odontogênicos/genética , Tumores Odontogênicos/patologia , Tumores Odontogênicos/metabolismo , Via de Sinalização Wnt/genética , Núcleo Celular/metabolismoRESUMO
BACKGROUND: Clear cell odontogenic carcinoma (CCOC) is an odontogenic carcinoma characterized by sheets and islands of vacuolated and clear cells. The diagnosis of atypical CCOC can pose a challenge when tumor cells deviate from their characteristic clear morphology, even with the aid of genetic profiling for CCOC identification. CASE PRESENTATION: In this manuscript, we detailed the inaugural instance of a recurrently recurring clear cell odontogenic carcinoma (CCOC) with pronounced squamous differentiation in a 64-year-old male. The primary tumor in this individual initially displayed a biphasic clear cell phenotype. However, subsequent to the third recurrence, the clear tumor cells were entirely supplanted by epidermoid cells characterized by eosinophilic cytoplasm, vesicular chromatin, and prominent nucleoli. Notable aggressive attributes such as necrosis, conspicuous cytological malignancy, perineural dissemination, and vascular invasion were noted. Additionally, the tumor progressed to manifest lung metastases. The tumor cells exhibited positive immunoreactivity for AE1/AE3, KRT19, Pan-CK, EMA, P40, P63, CK34ßE12, and P53, while they tested negative for CK35ßH11, KRT7, S-100, and neuroendocrine markers. The Ki-67 proliferation index was calculated at an average of 15%. Furthermore, FISH analysis unveiled the presence of the EWSR1::ATF1 gene fusion. CONCLUSIONS: This case illustrated a rare and aggressive case of CCOC characterized by significant squamous differentiation upon recurrence of the tumor.
Assuntos
Biomarcadores Tumorais , Tumores Odontogênicos , Humanos , Masculino , Pessoa de Meia-Idade , Tumores Odontogênicos/patologia , Tumores Odontogênicos/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/análise , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/genética , Adenocarcinoma de Células Claras/genética , Adenocarcinoma de Células Claras/patologia , Proteínas de Fusão Oncogênica/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/genética , Imuno-HistoquímicaRESUMO
Salivary gland and odontogenic neoplasms with extensive clear cell change are rare lesions but have been increasingly characterized over the past decade. Among this heterogeneous group of neoplasms, hyalinizing clear cell carcinoma (HCCC), clear cell odontogenic carcinoma (CCOC), and clear cell myoepithelial carcinoma (CCMC) share a monophasic clear cell morphology and an EWSR1 gene rearrangement. While HCCC is relatively well characterized, there are a limited number of EWSR1-reaarranged CCMC of salivary glands reported, and its clinicopathologic features in relation to HCCC and nonclear cell myoepithelial carcinoma (MC) have not been clarified. This report describes the clinical, morphologic, and immunophenotypic features of 3 carcinomas composed predominantly of clear cells and with EWSR1 rearrangements by fluorescence in situ hybridization. A comparative literature analysis suggests that HCCC, CCMC, and nonclear cell MC of salivary glands are clinically, histopathologically, and molecularly distinct.
Assuntos
Rearranjo Gênico , Hibridização in Situ Fluorescente , Proteína EWS de Ligação a RNA , Humanos , Proteína EWS de Ligação a RNA/genética , Masculino , Feminino , Pessoa de Meia-Idade , Neoplasias das Glândulas Salivares/genética , Neoplasias das Glândulas Salivares/patologia , Tumores Odontogênicos/genética , Tumores Odontogênicos/patologia , Adulto , Adenocarcinoma de Células Claras/genética , Adenocarcinoma de Células Claras/patologia , Idoso , Biomarcadores Tumorais/genética , Proteínas de Ligação a Calmodulina/genéticaRESUMO
OBJECTIVE: Central odontogenic fibromas (COF) are rare, benign tumors derived from dental mesenchymal tissue that may occur in the maxilla or mandible. This report describes primary and recurrent COF in the mandible of a patient with nevoid basal cell carcinoma syndrome (NBCCS). STUDY DESIGN: A 36-year-old African American male presented with a COF and its recurrence 17 months later. Tissue pieces were obtained from both occurrences with IRB-approved signed consent. Collected tissue pieces were dissected; one portion was formalin-fixed and paraffin-embedded, and the other was cultured for the isolation of cell populations from the primary (COdF-1) and recurrent (COdF-1a) tumors. Quantification real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, and DNA sequencing were used for gene and protein analysis of the primary tumor and cell populations. RESULTS: Histopathologic analysis of the tumor showed sparse odontogenic epithelial cords in fibrous connective tissue, and qRT-PCR analysis of tumor and cell populations (COdF-1 and COdF-1a) detected VIM, CK14, CD34, CD99 and ALPL mRNA expression. Protein expression was confirmed by immunohistochemistry. CD34 expression in primary tissues was higher than in tumor cells due to tumor vascularization. DNA sequencing indicated the patient had PTCH1 mutations. CONCLUSIONS: Histopathology, mRNA, and protein expression indicate the rare occurrence of COF in a patient with mutated PTCH1 gene and NBCCS.
Assuntos
Síndrome do Nevo Basocelular , Fibroma , Recidiva Local de Neoplasia , Tumores Odontogênicos , Humanos , Masculino , Síndrome do Nevo Basocelular/genética , Síndrome do Nevo Basocelular/patologia , Tumores Odontogênicos/patologia , Tumores Odontogênicos/genética , Tumores Odontogênicos/cirurgia , Adulto , Recidiva Local de Neoplasia/patologia , Fibroma/patologia , Fibroma/genética , Fibroma/cirurgia , Imuno-Histoquímica , Neoplasias Mandibulares/patologia , Neoplasias Mandibulares/genética , Neoplasias Mandibulares/cirurgia , Reação em Cadeia da Polimerase em Tempo Real , Técnicas In VitroRESUMO
Little is known about the histogenesis of the odontogenic myxoma (OM). Dental pulp stem cells could be candidate precursors of OM because both OM and the dental pulp share the same embryological origin: the dental papilla. For the purpose of comparing OM and stem cells, this study analyzed the expression of two proteins related to OM invasiveness (MMP-2 and hyaluronic acid) in human immature dental pulp stem cells (hIDPSCs). Three lineages of hIDPSCs from deciduous and permanent teeth were used in this study. Immunofluorescence revealed positive reactions for MMP-2 and hyaluronic acid (HA) in all hIDPSCs. MMP-2 appeared as dots throughout the cytoplasm, whereas HA appeared either as diffuse and irregular dots or as short fibrils throughout the cytoplasm and outside the cell bodies. The gene expression profile of each cell lineage was evaluated using RT-PCR analysis, and HA was expressed more intensively than MMP-2. HA expression was similar among the three hIDPSCs lineages, whereas MMP-2 expression was higher in DL-1 than in the other cell lines. The expression of proteins related to OM invasiveness in hIDPSCs could indicate that OM originates from dental pulp stem cells.
Assuntos
Adolescente , Criança , Pré-Escolar , Humanos , Polpa Dentária/metabolismo , Ácido Hialurônico/metabolismo , /metabolismo , Mixoma/patologia , Tumores Odontogênicos/patologia , Células-Tronco/metabolismo , Células Cultivadas , Polpa Dentária/citologia , Matriz Extracelular , Imunofluorescência , Expressão Gênica , Ácido Hialurônico/genética , /genética , Mixoma/genética , Invasividade Neoplásica/patologia , Tumores Odontogênicos/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
INTRODUÇÃO E OBJETIVO: O conhecimento do comportamento biológico de lesões de natureza odontogênica é essencial para tornar a abordagem terapêutica adequada e estabelecer um prognóstico. A produção de metaloproteinases da matriz extracelular (MMPs), a angiogênese e a proliferação celular fornecem subsídios para o crescimento tumoral. O presente artigo tem como objetivo fazer uma revisão de literatura de pesquisas em tumores odontogênicos (TOs) selecionados a partir da nova classificação da Organização Mundial da Saúde (OMS) de 2005 sobre a expressão de MMPs, marcadores angiogênicos e proliferação celular e verificar, nestes estudos, a relação desses marcadores quanto ao comportamento biológico dessas lesões. RESULTADOS: Nota-se que as MMPs -1, -2, -7, -9 e -26 encontram-se mais expressas no componente epitelial e estroma e, particularmente, a -13 em estroma. Uma maior angiogênese é observada em TOs mais agressivos. CD 105 foi mais expresso no TO ceratocístico (TOC) e CD34 em ameloblastomas sólidos (ASs). Relata-se elevada expressão do Ki-67 e p53 no TOC e no AS e baixo índice de proliferação celular no TO adenomatoide (TOA). CONCLUSÃO: Esses resultados mostram que as MMPs participam no processo de invasão e recorrência de algumas lesões odontogênicas, estando associadas ao comportamento biológico desses tumores. A angiogênese é fundamental para fornecer suporte à proliferação celular e esses dois eventos em conjunto estão correlacionados com diferentes níveis de comportamento biológico nos TOs, quando comparados com cistos de natureza odontogênica, o que pode sugerir o uso de inibidores angiogênicos como provável abordagem terapêutica nessas lesões.
INTRODUCTION AND OBJECTIVE: The study of biological behavior of odontogenic lesions is essential to the establishment of appropriate therapeutic approach and prognosis. The production of extracellular matrix metalloproteinases (MMPs), angiogenesis and cell proliferation contribute to tumor growth. This paper aims to review the literature on odontogenic tumors (OT) selected according to the new World Health Organization classification (WHO- 2005) by evaluating the expression of MMPs, angiogenic and cell proliferation. Furthermore, it aims to verify the relation between these markers and the biological behavior of these lesions. RESULTS: it was found that MMPs -1, -2, -7, -9 and -26 had a higher expression in both epithelial component and stroma, and 13 particularly in the stroma. Increased angiogenesis was observed in more aggressive OT. CD105 expression was higher in keratocystic odontogenic tumour (KOT) and CD34 in solid ameloblastomas (SA). It was observed a higher expression of Ki-67 and p53 in SA and KOT and a low cell proliferation rate in the adenomatoid odontogenic tumour (AOT). CONCLUSION: These results show that MMPs are involved in invasion and recurrence of some odontogenic lesions and are associated with the biological behavior of these tumors. Angiogenesis is critical to provide support to cell proliferation and these concomitant events are correlated with different levels of biological behavior in OT when compared to odontogenic cysts, hence the use of angiogenic inhibitors may be a potential therapeutic approach in these lesions.
Assuntos
Matriz Extracelular , Metaloproteinases da Matriz Associadas à Membrana/genética , Proliferação de Células , Tumores Odontogênicos/genéticaRESUMO
Odontogenic myxoma (OM) is an ectomesenchymal benign odontogenic tumor characterized by spindle or stellate-shaped cells embedded in an abundant myxoid or mucoid extracellular matrix. DNA methylation is characterized by the addition of methyl groups in cytosines within CpG islands in the promoter gene. DNA methylation can decrease the expression of tumor suppressor genes and contribute to the development of neoplastic lesions. The aim of study was to evaluate the methylation pattern of the tumor suppressor genes P16 (CDKN2A), P21 (CDKN1A), P27 (CDKN1B), P53 (TP53) and RB1 in OM and dental pulp. Methylation was evaluated using methylation-specific polymerase chain reaction (PCR). The transcription was studied in some cases by using reverse transcription quantitative PCR. A higher frequency of unmethylated P27, P53, and RB1 samples was observed in the OM when compared with the dental pulp. OM expressed mRNA of all the genes evaluated. Considering all the samples together, the expression of Rb was higher in the unmethylated samples compared with the partially methylated samples. This investigation revealed hypomethylation of the genes P27, P53, and RB1 in OM. In addition, methylation of tumor suppressor genes was found to be an usual event in normal dental pulp.
O mixoma odontogênico (MO) é um tumor odontogênico benigno de origem mesenquimal caracterizado pela presença de células fusiformes ou estreladas dispostas em abundante matriz extracelular mucóide. A metilação do DNA é caracterizada pela adição de grupos metil em citosinas constituintes de ilhas CpG na região promotora do gene. A metilação pode diminuir a expressão de genes supressores de tumor e contribuir para o desenvolvimento de lesões neoplásicas. O objetivo deste trabalho foi avaliar o padrão de metilação nos genes P16 (CDKN2A), P21 (CDKN1A), P27 (CDKN1B), P53 (TP53), RB1 nos MO e na polpa dental. A metilação foi avaliada pela reação em cadeia da polimerase específica para a metilação. A transcrição dos genes foi estudada em alguns casos pela reação da transcriptase reversa (PCR quantitativa). Uma maior frequência de amostras não metiladas para os genes P27, P53 e RB1 foi observada nos MO quando comparados à polpa dental. Os MO expressaram RNAm de todos os genes avaliados. Considerando todas as amostras juntas, a expressão de Rb foi maior em amostras não metiladas comparadas as amostras parcialmente metiladas. Esta investigação mostrou a hipometilação dos genes P27, P53 e RB1 nos MO. Adicionalmente, a metilação nos genes supressores de tumor é um evento frequente em polpa dental normal.
Assuntos
Adolescente , Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Metilação de DNA/genética , Genes Supressores de Tumor/fisiologia , Tumores Odontogênicos/genética , Citosina , Ilhas de CpG/genética , /genética , /genética , Polpa Dentária/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , /fisiologia , /genética , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/genética , Proteína do Retinoblastoma/genética , Transcrição Gênica/genéticaRESUMO
Os tumores odontogênicos são um grupo heterogéneo de lesões formadas a partir de tecidos que dão origem ao dente. A metilação do ADN, uma adição covalente de um grupo metilo na posição 5 de carbono de um nucleótideo de citosina, é considerado um importante regulador da expressão génica. A adição do radical metil é catalisada por ADN metiltransferases (DNMTs). Embora alguns estudos epigenéticos tenham sido realizados em tumores odontogênicos, um estudo com os três tipos de DNMTs em vários membros desse grupo está em falta. Este estudo analisa a expressão de DNMTs em tumores odontogênicos. Amostras de vinte ameloblastomas, dez Calcificante tumores odontogênicos císticos, dez calcificados tumores epiteliais, dez tumor odontogênico adenomatóide, dez tumores odontogênicos queratocísticos, quatro fibromas ameloblásticos, dois fibro-odontoma ameloblástico, quatro fibroma centrais odontogênicos, sete tecidos de fibromas odontogênicos periféricos e dez mixomas odontogênicos foram incluídos. DNMT1, 3A e 3B foram expressas no núcleo e / ou citoplasma de todos os tumores odontogênicos. A alta expressão de DNMTs em células de tumor odontogênico sugere metilação como um mecanismo importante para este grupo de tumores.
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 catalyzed 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 analyzes the expression of DNMTs in odontogenic tumours. Formalin-fixed and paraffin-embedded tissue samples of twenty ameloblastomas, ten calcifying cystic odontogenic tumors, ten calcifying epithelial tumors, ten adenomatoid odontogenic tumors, ten keratocystic odontogenic tumors, five ameloblastic fibromas, two ameloblastic fibro-odontoma, four central odontogenic fibroma, seven peripheral odontogenic fibroma and ten odontogenic mixoma were included. DNMT1, 3A and 3B were expressed in the nucleus and/or cytoplasm of all odontogenic tumours. The high expression of DNMTs in odontogenic tumour cells suggests methylation as an important mechanism for this group of tumours.