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1.
J Mol Diagn ; 2020 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-32966885

RESUMO

The molecular pathogenesis of mixed odontogenic tumors has not been established and understanding their genetic basis could refine their classification and help define molecular markers for diagnostic purposes. We assessed potentially pathogenic mutations in the component tissues of 28 cases of mixed odontogenic tumors. Laser capture microdissected tissue from ten ameloblastic fibromas (AF), four ameloblastic fibrodentinomas (AFD), six ameloblastic fibro-odontomas (AFO), three ameloblastic fibrosarcomas (AFS), and five odontomas (OD) were screened by next-generation sequencing for a panel of 50 oncogenes and tumour suppressor genes and results confirmed by TaqMan allele-specific qPCR. We showed BRAF p.V600E mutation in the mesenchymal component of 4/10 (40%) AF, 2/4 (50%) AFD, 2/6 (33%) AFO, and 2/3 (67%) AFS, while all five OD were wild-type for BRAF p.V600E. Mutation in the epithelial component was only observed in one AF and one AFO. One AFS contained an area of benign AF and the mesenchymal component of both (AFS and AF) contained BRAF p.V600E, supporting the concept of malignant progression from a benign AF precursor. We report KDR, TP53, KIT and PIK3CA SNPs. In conclusion, AF, AFD, AFO and AFS show BRAF p.V600E in their mesenchymal component unlike OD, which are BRAF wild-type, suggesting that at least a subset of AF, AFD and AFO are molecularly distinct from OD, and may represent distinct entities and be neoplastic.

2.
Oral Dis ; 2020 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-32772410

RESUMO

OBJECTIVE: Odontogenic keratocyst (OKC) is a benign lesion that tends to recur after surgical treatment. In an attempt to clarify the molecular basis underlining the OKC pathobiology, we aimed to analyze its proteomic profile. MATERIALS AND METHODS: We compared the proteomic profiles of five OKC and matched normal oral mucosa by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Then, we performed enrichment analysis and a literature search for the immunoexpression of the proteomics targets. RESULTS: We identified 1,150 proteins and 72 differently expressed proteins (log2 fold change ≥ 1.5; p < .05). Twenty-seven peptides were exclusively detected in the OKC samples. We found 35 enriched pathways related to cell differentiation and tissue architecture, including keratinocyte differentiation, keratinization, desmosome, and extracellular matrix (ECM) organization and degradation. The immunoexpression information of 11 out of 50 proteins identified in the enriched pathways was obtained. We found the downregulation of four desmosomal proteins (JUP, PKP1, PKP3, and PPL) and upregulation of ECM proteases (MMP-2, MMP-9, and cathepsins). CONCLUSIONS: Proteomic analysis strengthened the notion that OKC cells have a similar proteomic profile to oral keratinocytes. Contextual investigation of the differentially expressed proteins revealed the deregulation of desmosome proteins and ECM degradation as important alterations in OKC pathobiology.

3.
Arch Oral Biol ; 113: 104523, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31543246

RESUMO

OBJECTIVE: Driver oncogenic mutations have been reported in several benign neoplasms. While ameloblastomas show BRAF p.V600E mutations, adenomatoid odontogenic tumours harbour either KRAS p.G12R or p.G12 V. The lack of understanding of the core molecular changes involved in tumour initiation and progression represents a critical barrier to developing new strategies for cancer detection and prevention. Considering the fact that ameloblastoma and adenomatoid odontogenic tumours can originate from dental follicles, we hypothesized that the BRAF and KRAS mutations might be early events in odontogenic tumours tumourigenesis. We aimed to assess BRAF and KRAS mutations in dental follicles associated with asymptomatic impacted teeth. DESIGN: Forty-eight dental follicles containing odontogenic epithelial remnants were included in the study. As ameloblastomas most often occur in the posterior mandible and adenomatoid odontogenic tumours have a predilection for the anterior jaws, we assessed by allele-specific qPCR the presence of BRAF p.V600E in 32 dental follicles associated with impacted 3rd mandibular molar teeth and KRAS p.G12 V and KRAS p.G12R mutations in 16 dental follicle specimens obtained from around impacted anterior teeth. Sanger sequencing was used as an additional method. RESULTS: None of the dental follicle cases tested positive for the mutations. CONCLUSION: In conclusion, we tried to detect the early genetic events associated with odontogenic tumours development in dental follicles, but we were unable to showcase that BRAF p.V600E and KRAS p.G12R or p.G12 V mutations are the early genetic events associated with odontogenic tumours development.

5.
Clin Oral Investig ; 2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31713744

RESUMO

OBJECTIVE: Odontogenic myxoma (OM) occasionally responds poorly to surgical treatment. The MAPK pathway is constitutively activated in several neoplasms and we aimed to test if the MAPK pathway is activated in OM, in order to pave the way for an alternative therapy for aggressive and recurrent cases. MATERIALS AND METHODS: The immunoexpression of phosphorylated ERK1/2 (pERK1/2) was assessed in OM. We established a 3D organotypic culture model for the in vitro study and patient-derived xenografts (PDX) in mice for the in vivo study. The MEK inhibitor U0126 was used to inhibit phosphorylation of ERK1/2 in the in vitro and in vivo models. RESULTS: All OM showed strong pERK1/2 immunoexpression, consistent with MAPK pathway activation. Treatment of the 3D culture with U0126 resulted in a reduced pERK1/2/ERK1/2 ratio. Consistent with the in vitro results, all PDX of animals treated with U0126 showed a decreased volume fold change compared with controls. CONCLUSIONS: The MAPK pathway is activated in OM and its inhibition leads to tumor shrinkage in PDX and cell culture models. CLINICAL RELEVANCE: Our results offer a pre-clinical frame for OM-targeted therapy. Further work is needed to determine if this initial finding holds clinical promise.

6.
J Oral Pathol Med ; 48(10): 906-910, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31310691

RESUMO

BACKGROUND: Pyogenic granuloma (PG) is a benign nodular lesion with a prominent vascular component which may affect different sites. Recently, BRAF, KRAS, HRAS, NRAS, GNA11, and GNA14 mutations were reported on PGs of the skin. The present study assessed the role of the MAPK/ERK pathway in oral PG pathogenesis. METHODS: Mutations in hotspot regions of genes involved in the MAPK/ERK pathway activation were investigated by Sanger sequencing. The expression of phospho-ERK1/2 was evaluated by immunohistochemistry. RESULTS: Oral PGs did not show mutations in the sequenced regions of the genes BRAF, KRAS, HRAS, NRAS, GNA11, or GNA14. Our results also showed activation of the MAPK/ERK pathway demonstrated by phospho-ERK1/2 immunohistochemical positivity. CONCLUSIONS: Although oral PG shows MAPK/ERK pathway activation, the driver molecular event remains to be elucidated.


Assuntos
Granuloma Piogênico/metabolismo , Sistema de Sinalização das MAP Quinases , Mutação , Adolescente , Adulto , Idoso , Feminino , GTP Fosfo-Hidrolases/metabolismo , Subunidades alfa de Proteínas de Ligação ao GTP/metabolismo , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Granuloma Piogênico/genética , Humanos , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas B-raf/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Transdução de Sinais , Adulto Jovem
7.
J Oral Pathol Med ; 48(5): 421-423, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30849196

RESUMO

Chronic mucosal trauma is suggested as an additional etiologic risk factor for oral squamous cell carcinoma (OSCC), but there is a lack of experimental-molecular data. If chronic trauma of the oral mucosa is carcinogenic, it should be associated with early genetic alterations seen during typical progression of OSCC, like loss of heterozygosity (LOH). We investigated LOH in the key chromosomal arms 3p, 9p and 17p in inflammatory fibrous hyperplasia associated with removable dental prosthesis and also in normal oral mucosa, by using the polymorphic microsatellite markers D3S1300 at 3p14.2, D9S1748 at 9p21, D17S1289 at 17p12 and D17S974 at 17p13 and capillary electrophoresis. LOH was detected in 2/15 (13%) fibrous hyperplasia samples similarly to other reactive and inflammatory lesions. None of the normal mucosa samples presented LOH. Our experimental-molecular results do not support the hypothesis that trauma associated with dental prosthesis has an important role in oral carcinogenesis.


Assuntos
Carcinoma de Células Escamosas/complicações , Dentaduras/efeitos adversos , Perda de Heterozigosidade , Neoplasias Bucais/complicações , Boca/lesões , Adulto , Idoso , Carcinogênese , Cromossomos Humanos Par 17 , Cromossomos Humanos Par 3 , Cromossomos Humanos Par 9 , Feminino , Humanos , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade
8.
J Oral Pathol Med ; 48(5): 400-405, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30801783

RESUMO

BACKGROUND: Oral Lichen Planus is a chronic inflammatory disorder that affects the oral mucosa, with the reticular and erosive forms representing the primary clinical variants of the disease. Previous studies have shown that metabolic alterations may well be involved in the pathogenesis of the disease; however, the molecular mechanisms related to the clinicopathological differences between erosive and reticular forms remain unknown. METHODS: A comparative metabolomic analysis was performed on formalin-fixed and paraffin-embedded tissue samples of erosive (n = 6) and reticular (n = 10) oral lichen planus using gas chromatography-mass spectrometry. RESULTS: The metabolomic analysis showed a distinct profile between the two clinical variants. Five metabolites (cyclohexanamine, glycine, mannitol/sorbitol, methyl palmitate and trehalose) were significantly diminished in erosive oral lichen planus as compared to the reticular form. CONCLUSIONS: Reticular and erosive forms of oral lichen planus have a distinct metabolic profile. However, further studies using a large number of fresh tissue samples are necessary to confirm this data.


Assuntos
Líquen Plano Bucal/classificação , Líquen Plano Bucal/metabolismo , Metaboloma , Adulto , Idoso , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Líquen Plano Bucal/patologia , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/metabolismo , Mucosa Bucal/patologia , Adulto Jovem
9.
J Oral Pathol Med ; 48(4): 307-314, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30739334

RESUMO

BACKGROUND: Ameloblastoma is a locally infiltrative, aggressive epithelial odontogenic neoplasm. BRAF-V600E mutation is frequently found in this tumor and has a pivotal role in its pathogenesis, but the consequences of this alteration need to be addressed. An untargeted metabolomics approach was applied to verify whether metabolic disturbances are related to tumor biology and whether BRAF-V600E mutation contributes to these alterations. METHODS: Formalin-fixed and paraffin-embedded tissue specimens from thirteen ameloblastoma and six dental follicles were included in this study. BRAF mutational status was determined by competitive allele-specific real-time PCR. Metabolite extracts were analyzed using gas chromatography coupled to mass spectrometry. Univariate and multivariate statistical methods were employed to compare the metabolic profiles of the samples. RESULTS: The abundance of eleven metabolites was significantly higher in ameloblastoma in relation to dental follicles, including amino acids, fatty acids, carbohydrates, inorganic acids, and organoheterocyclic compounds. The presence of BRAF-V600E mutations in ameloblastoma was related to decreased levels of glycerol in comparison with tumors carrying only wild-type alleles of this gene. No metabolic differences were observed between recurrent and primary manifestations of ameloblastoma. CONCLUSIONS: Ameloblastoma exhibits a distinct metabolic profile from normal odontogenic epithelium. BRAF-V600E may contribute to metabolic alterations in ameloblastoma. Collectively, our findings suggest that metabolic alterations might play a role in tumor pathogenesis.


Assuntos
Ameloblastoma/genética , Tumores Odontogênicos/genética , Proteínas Proto-Oncogênicas B-raf/genética , Alelos , Ameloblastoma/metabolismo , Análise Mutacional de DNA , Humanos , Mutação , Tumores Odontogênicos/metabolismo
10.
Mod Pathol ; 32(6): 799-806, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30643167

RESUMO

Adenomatoid odontogenic tumor is a benign encapsulated epithelial odontogenic tumor that shows an indolent clinical behavior. We have reported in a few adenomatoid odontogenic tumors mutations in KRAS, which is a proto-oncogene frequently mutated in cancer such as lung, pancreas, and colorectal adenocarcinomas. We aimed to assess KRAS mutations in the hotspot codons 12, 13, and 61 in a large cohort of adenomatoid odontogenic tumors and to test the association of these mutations with clinical (age, site, tumor size, follicular/extrafollicular subtypes) and histopathological parameters. Thirty eight central cases were studied. KRAS codon 12 mutations were assessed by TaqMan allele-specific qPCR (p.G12V/R) and/or Sanger sequencing, and codon 13 and 61 mutations were screened by Sanger. Histological tumor capsule thickness was evaluated by morphometric analysis. Additionally, the phosphorylated form of the MAPK downstream effector ERK1/2 was investigated. Statistical analysis was carried out to test the association of KRAS mutations with clinicopathological parameters. KRAS c.35 G >T mutation, leading to p.G12V, was detected in 15 cases. A novel mutation in adenomatoid odontogenic tumor, c.34 G >C, leading to p.G12R, was detected in 12 cases and the other 11 were wild-type. Codon 12 mutations were not associated with the clinicopathological parameters tested. RAS mutations are known to activate the MAPK pathway, and we show that adenomatoid odontogenic tumors express phosphorylated ERK1/2. In conclusion, a high proportion of adenomatoid odontogenic tumors (27/38, 71%) have KRAS codon 12 mutations, which occur independently of the clinicopathological features evaluated. Collectively, these findings indicate that KRAS mutations and MAPK pathway activation are the common features of this tumor and some cancer types. Although it is unclear why different codon 12 alleles occur in different disease contexts and the complex interactions between tumor genotype and phenotype need clarification, on the basis of our results the presence of KRAS p.G12V/R favors the adenomatoid odontogenic tumor diagnosis in challenging oral neoplasm cases.


Assuntos
Ameloblastoma/genética , Ameloblastoma/patologia , Proteínas Proto-Oncogênicas p21(ras)/genética , Adolescente , Adulto , Criança , Feminino , Humanos , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Pessoa de Meia-Idade , Mutação , Adulto Jovem
11.
Pathol Res Pract ; 215(3): 466-469, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30595406

RESUMO

Ameloblastoma and adenomatoid odontogenic tumor (AOT) are jaw tumors derived from the teeth forming apparatus. While ameloblastoma is a destructive, debilitating lesion, with conventional surgical treatment leading to facial deformity and morbodities, AOT shows indolent clinical behavior. The underlying molecular mechanisms associated with their biological behavior are unknown. The use of high-density whole-genome microarray analysis in ameloblastomas and AOT revealed high frequency of genomic gain at 14q32.33, which encompasses the long noncoding RNA (lncRNA) gene KIAA0125. In the present study, we aimed to investigate the expression profile of KIAA0125 in these tumors. Thirteen samples were included (five solid/multicystic ameloblastomas, four AOT, and four dental follicles). The relative quantification of KIAA0125 expression was obtained by qPCR and interactions of KIAA0125 were in silico predicted. We detected higher levels of KIAA0125 transcripts in the ameloblastoma group compared to dental follicles (p = 0.042). The expression levels of KIAA0125 in AOT were not different from that of dental follicles. KIAA0125 was predicted to interact with 41 miRNA families. Four miRNAs of these families have been previously reported differentially expressed in ameloblastoma, being miR-135a-5p, miR-204-5p and miR-205-5p upregulated, and miR-150-5p downregulated. The lncRNA KIAA0125 is likely involved in the ameloblastoma pathobiology. LncRNAs hold strong promise as therapeutic targets and experimental validation of this lncRNA functions may lead to tailored therapies targeting KIAA0125 in extensive and recurrent ameloblastoma cases.


Assuntos
Ameloblastoma/genética , Neoplasias Maxilomandibulares/genética , RNA Longo não Codificante/biossíntese , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Regulação para Cima
12.
J Oral Pathol Med ; 48(2): 174-179, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30554445

RESUMO

BACKGROUND: Benign neoplasms exhibit most of the cellular phenomena considered hallmarks of cancer, except the capacity to metastasize. Thus, the elucidation of the mechanisms associated with the progression of benign neoplasms may complement and clarify the mechanisms involved in carcinogenesis. Benign odontogenic tumours often result in facial deformities and morbidities, and have complex pathogenesis, mainly due to the diversity of interactions between the odontogenic epithelium and the ectomesenchyme. Primary cell culture of such tumours is not only difficult to be established and maintained, but also tumour cells lose characteristic cellular morphology. Considering gene expression, growth, migration, proliferation and cellular morphology are controlled by cell-cell interactions and cell-extracellular matrix interactions, cell culture in 3D substrates has gained space as a way to overcome some of the limitations of traditional monolayer cell culture systems. METHODS: In this study, fragments obtained from mesenchymal odontogenic tumours were cultured in type I collagen scaffolds. Invasion tests were performed in these models, as well as phenotypic characterization of the cultured tumours. RESULTS: The results obtained for the odontogenic myxoma and the cemento-ossifying fibroma demonstrate a good reproduction of the growth pattern of these tumours under ex vivo conditions. Microscopic evaluation showed maintenance of cell viability in the explants for more than 30 days, without the presence of necrosis. CONCLUSION: This is the first study involving long-term 3D primary cultures of benign odontogenic tumours, which is expected to support complex approaches to cell and molecular biology, and to serve as an experimental model for testing molecular therapies.


Assuntos
Técnicas de Cultura de Células/métodos , Técnicas In Vitro , Tumores Odontogênicos/patologia , Carcinogênese , Comunicação Celular , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Cementoma , Expressão Gênica , Humanos , Tumores Odontogênicos/genética , Células Tumorais Cultivadas
13.
Nat Commun ; 9(1): 4572, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30385747

RESUMO

Giant cell lesions of the jaw (GCLJ) are debilitating tumors of unknown origin with limited available therapies. Here, we analyze 58 sporadic samples using next generation or targeted sequencing and report somatic, heterozygous, gain-of-function mutations in KRAS, FGFR1, and p.M713V/I-TRPV4 in 72% (42/58) of GCLJ. TRPV4 p.M713V/I mutations are exclusive to central GCLJ and occur at a critical position adjacent to the cation permeable pore of the channel. Expression of TRPV4 mutants in HEK293 cells leads to increased cell death, as well as increased constitutive and stimulated channel activity, both of which can be prevented using TRPV4 antagonists. Furthermore, these mutations induce sustained activation of ERK1/2, indicating that their effects converge with that of KRAS and FGFR1 mutations on the activation of the MAPK pathway in GCLJ. Our data extend the spectrum of TRPV4 channelopathies and provide rationale for the use of TRPV4 and RAS/MAPK antagonists at the bedside in GCLJ.


Assuntos
Tumor de Células Gigantes do Osso/genética , Neoplasias Maxilomandibulares/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/genética , Canais de Cátion TRPV/genética , Adolescente , Adulto , Idoso , Criança , Simulação por Computador , Feminino , Mutação com Ganho de Função , Células HEK293 , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Sistema de Sinalização das MAP Quinases , Masculino , Pessoa de Meia-Idade , Técnicas de Patch-Clamp , Análise de Sequência de DNA , Análise de Sequência de RNA , Sequenciamento Completo do Exoma , Adulto Jovem
14.
J Oral Pathol Med ; 47(1): 91-95, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28976032

RESUMO

BACKGROUND: The oral lichen planus is a chronic inflammatory disease. Although its aetiology is not well understood, the role of T lymphocytes in its inflammatory events is recognised. Identifying the epigenetic mechanisms involved in the pathogenesis of this immune-mediated condition is fundamental for understanding the inflammatory reaction that occurs in the disease. The purpose of this work was to evaluate the methylation pattern of 21 immune response-related genes in the different clinical forms of oral lichen planus. METHODS: A cross-sectional study was performed to analyse the DNA methylation patterns in three distinct groups of oral lichen planus: (i) reticular/plaque lesions; (ii) erosive lesions; (iii) normal oral mucosa (control group). After DNA extraction from biopsies, the samples were submitted to digestions by methylation-sensitive and methylation-dependent enzymes and double digestion. The relative percentage of methylated DNA for each gene was provided using real-time polymerase chain reaction arrays. RESULTS: Hypermethylation of the STAT5A gene was observed only in the control group (59.0%). A higher hypermethylation of the ELANE gene was found in reticular/plaque lesions (72.1%) compared to the erosive lesions (50.0%). CONCLUSION: Our results show variations in the methylation profile of immune response-related genes, according to the clinical type of oral lichen planus after comparing with the normal oral mucosa. Further studies are necessary to validate these findings using gene expression analysis.


Assuntos
Metilação de DNA , Epigenômica , Líquen Plano Bucal/genética , Líquen Plano Bucal/imunologia , Adolescente , Adulto , Idoso , Estudos Transversais , DNA/análise , DNA/isolamento & purificação , Feminino , Expressão Gênica , Humanos , Inflamação , Líquen Plano Bucal/patologia , Masculino , Pessoa de Meia-Idade , Mucosa Bucal , Regiões Promotoras Genéticas , Fator de Transcrição STAT5/genética , Linfócitos T , Proteínas Supressoras de Tumor/genética , Adulto Jovem
15.
J Oral Pathol Med ; 47(1): 78-85, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29032608

RESUMO

BACKGROUND: Cemento-ossifying fibroma (COF) is a benign fibro-osseous neoplasm of uncertain pathogenesis, and its treatment results in morbidity. MicroRNAs (miRNA) are small non-coding RNAs that regulate gene expression and may represent therapeutic targets. The purpose of the study was to generate a comprehensive miRNA profile of COF compared to normal bone. Additionally, the most relevant pathways and target genes of differentially expressed miRNA were investigated by in silico analysis. METHODS: Nine COF and ten normal bone samples were included in the study. miRNA profiling was carried out by using TaqMan® OpenArray® Human microRNA panel containing 754 validated human miRNAs. We identified the most relevant miRNAs target genes through the leader gene approach, using STRING and Cytoscape software. Pathways enrichment analysis was performed using DIANA-miRPath. RESULTS: Eleven miRNAs were downregulated (hsa-miR-95-3p, hsa-miR-141-3p, hsa-miR-205-5p, hsa-miR-223-3p, hsa-miR-31-5p, hsa-miR-944, hsa-miR-200b-3p, hsa-miR-135b-5p, hsa-miR-31-3p, hsa-miR-223-5p and hsa-miR-200c-3p), and five were upregulated (hsa-miR-181a-5p, hsa-miR-181c-5p, hsa-miR-149-5p, hsa-miR-138-5p and hsa-miR-199a-3p) in COF compared to normal bone. Eighteen common target genes were predicted, and the leader genes approach identified the following genes involved in human COF: EZH2, XIAP, MET and TGFBR1. According to the biology of bone and COF, the most relevant KEGG pathways revealed by enrichment analysis were proteoglycans in cancer, miRNAs in cancer, pathways in cancer, p53-, PI3K-Akt-, FoxO- and TGF-beta signalling pathways, which were previously found to be differentially regulated in bone neoplasms, odontogenic tumours and osteogenesis. CONCLUSION: miRNA dysregulation occurs in COF, and EZH2, XIAP, MET and TGFBR1 are potential targets for functional analysis validation.


Assuntos
Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Fibroma Ossificante/genética , Fibroma Ossificante/metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , MicroRNAs/genética , Adolescente , Adulto , Biologia Computacional , Regulação para Baixo , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Feminino , Fatores de Transcrição Forkhead/metabolismo , Estudos de Associação Genética , Humanos , Masculino , MicroRNAs/classificação , Pessoa de Meia-Idade , Proteínas de Neoplasias/metabolismo , Tumores Odontogênicos , Osteogênese , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas c-met/genética , RNA não Traduzido , Receptor do Fator de Crescimento Transformador beta Tipo I , Receptores de Fatores de Crescimento Transformadores beta/genética , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/genética , Adulto Jovem
16.
J Clin Pathol ; 71(3): 279-283, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29127140

RESUMO

AIMS: To identify calcifying epithelial odontogenic tumour (CEOT) mutations in oncogenes and tumour suppressor genes. METHODS: A panel of 50 genes commonly mutated in cancer was sequenced in CEOT by next-generation sequencing. Sanger sequencing was used to cover the region of the frameshift deletion identified in one sample. RESULTS: Missense single nucleotide variants (SNVs) with minor allele frequency (MAF) <1% were detected in PTEN, MET and JAK3. A frameshift deletion in CDKN2A occurred in association with a missense mutation in the same gene region, suggesting a second hit in the inactivation of this gene. APC, KDR, KIT, PIK3CA and TP53 missense SNVs were identified; however, these are common SNVs, showing MAF >1%. CONCLUSION: CEOT harbours mutations in the tumour suppressor PTEN and CDKN2A and in the oncogenes JAK3 and MET. As these mutations occurred in only one case each, they are probably not driver mutations for these tumours.


Assuntos
Tumores Odontogênicos/genética , Oncogenes/genética , Neoplasias Cutâneas/genética , Proteínas Supressoras de Tumor/genética , Adolescente , Adulto , Inibidor p16 de Quinase Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p18 , Feminino , Frequência do Gene , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Janus Quinase 3/genética , Masculino , Proteínas de Membrana/genética , Microdissecção , Pessoa de Meia-Idade , Mutação , Mutação de Sentido Incorreto , Tumores Odontogênicos/patologia , PTEN Fosfo-Hidrolase/genética , Proteínas Proto-Oncogênicas c-met/genética , Análise de Sequência de DNA , Neoplasias Cutâneas/patologia
18.
J Oral Pathol Med ; 47(2): 186-191, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29272070

RESUMO

BACKGROUND: Mutations in the patched 1 (PTCH1) gene are the main genetic alteration reported in sporadic and nevoid basal cell carcinoma-associated odontogenic keratocyst (OKC). Oncogenic mutations, including BRAFV600E, previously considered exclusive of malignant neoplasms have been reported in odontogenic tumors. Recently, a high frequency of BRAFV600E mutation has been reported in OKC. Because of the considerable recurrence rate of OKC, the identification of druggable genetic mutations can be relevant in the management of extensive lesions. METHODS: A set of 28 OKCs was included in this work. Initially, 10 sporadic and eight OKC samples from four NBCCS patients (a pair of lesions from each syndromic patient) were submitted to targeted next-generation sequencing (NGS) of 2800 different mutations in 50 oncogenes and tumor suppressor genes, including BRAF. Ten extra sporadic OKC samples were included to assess BRAFV600E mutation using TaqMan allele-specific qPCR. RESULTS: The following missense mutations occurred in one case each: ATM p.Ser333Phe, SMO p.Gly416Glu, PIK3CA p.Ser326Phe, FBXW7 p.Ser438Phe, JAK2 p.Ser605Phe, PTEN p.Arg173His, ATM p.Cys353Arg, PTEN p.Ser294Arg, MET p.His1112Tyr. None of the 18 samples showed the BRAFV600E (or any other V600) mutation in the NGS. BRAFV600E mutation was detected by qPCR in one of the 10 OKC. Collectively, our results show BRAFV600E mutation in 1 of 28 OKC cases. CONCLUSION: On the basis of our results, OKCs do not present recurrent hotspot mutations in these 50 genes commonly mutated in cancer. In addition, BRAFV600E does not play a central role in OKC pathogenesis.


Assuntos
Síndrome do Nevo Basocelular/genética , Mutação , Cistos Odontogênicos/genética , Receptores de Superfície Celular/genética , Adolescente , Adulto , Idoso , Brasil , Carcinoma/genética , Criança , DNA/genética , DNA/isolamento & purificação , Feminino , Genes Supressores de Tumor , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Tumores Odontogênicos , Receptor Patched-1/genética , Adulto Jovem
19.
Artigo em Inglês | MEDLINE | ID: mdl-29239811

RESUMO

OBJECTIVE: The molecular pathogenesis of cemento ossifying fibroma (COF) is unclear. The purpose of this study was to investigate mutations in 50 oncogenes and tumor suppressor genes, including APC and CTNNB1, in which mutations in COF have been previously reported. In addition, we assessed the transcriptional levels of the Wnt/ß-catenin pathway genes in COF. STUDY DESIGN: We used a quantitative polymerase chain reaction array to evaluate the transcriptional levels of 44 Wnt/ß-catenin pathway genes in 6 COF samples, in comparison with 6 samples of healthy jaws. By using next-generation sequencing (NGS) in 7 COF samples, we investigated approximately 2800 mutations in 50 genes. RESULTS: The expression assay revealed 12 differentially expressed Wnt/ß-catenin pathway genes in COF, including the upregulation of CTNNB1, TCF7, NKD1, and WNT5 A, and downregulation of CTNNBIP1, FRZB, FZD6, RHOU, SFRP4, WNT10 A, WNT3 A, and WNT4, suggesting activation of the Wnt/ß-catenin signaling pathway. NGS revealed 5 single nucleotide variants: TP53 (rs1042522), PIK3 CA (rs2230461), MET (rs33917957), KIT (rs3822214), and APC (rs33974176), but none of them was pathogenic. CONCLUSIONS: Although NGS detected no oncogenic mutation, deregulation of key Wnt/ß-catenin signaling pathway genes appears to be relevant to the molecular pathogenesis of COF.


Assuntos
Fibroma Ossificante/genética , Neoplasias Mandibulares/genética , Via de Sinalização Wnt/genética , Adulto , Regulação para Baixo , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Mutação/genética , Reação em Cadeia da Polimerase , Ativação Transcricional
20.
Artigo em Inglês | MEDLINE | ID: mdl-28941993

RESUMO

OBJECTIVE: The aim of this study was to evaluate the DNA methylation profile in 22 apoptosis-related genes in pleomorphic adenomas (PAs) of the salivary glands, in comparison with normal salivary glands (NSGs), and to address the differences in methylation patterns between smaller and larger tumors. Additionally, we investigated if the hypermethylation of differentially methylated genes between NSGs and PAs impacted the messenger RNA (mRNA) transcription. DESIGN: Twenty-three fresh PA samples and 12 NSG samples were included. The PA samples were divided into 2 groups: PAs with clinical size larger than 2 cm (n = 12) and PAs with clinical size 2 cm or smaller (n = 11). DNA methylation at the promoter region of a panel of 22 genes involved in apoptosis was profiled by using a human apoptosis DNA methylation polymerase chain reaction array, and the transcriptional levels of genes showing differential methylation profiles between PAs and NSGs were assessed. RESULTS: TNFRSF25 and BCL2 L11 were highly methylated in PAs, in comparison with NSGs, irrespective of tumor size. However, no difference could be observed in the mRNA transcription between PAs and NSGs. CONCLUSIONS: Hypermethylation of the proapoptotic genes BCL2 L11 and TNFRSF25 is observed in PA. However, this phenomenon did not impact mRNA transcription.


Assuntos
Adenoma Pleomorfo/genética , Proteína 11 Semelhante a Bcl-2/genética , Metilação de DNA , Reação em Cadeia da Polimerase , Membro 25 de Receptores de Fatores de Necrose Tumoral/genética , Neoplasias das Glândulas Salivares/genética , Adenoma Pleomorfo/patologia , Adulto , Idoso , Apoptose , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias das Glândulas Salivares/patologia
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