RESUMO
The study sought to identify genetic aberrations driving oral squamous cell carcinoma (OSCC) development among users of shammah, an Arabian preparation of smokeless tobacco. Twenty archival OSCC samples, 15 of which with a history of shammah exposure, were whole-exome sequenced at an average depth of 127×. Somatic mutations were identified using a novel, matched controls-independent filtration algorithm. CODEX and Exomedepth coupled with a novel, Database of Genomic Variant-based filter were employed to call somatic gene-copy number variations. Significantly mutated genes were identified with Oncodrive FM and the Youn and Simon's method. Candidate driver genes were nominated based on Gene Set Enrichment Analysis. The observed mutational spectrum was similar to that reported by the TCGA project. In addition to confirming known genes of OSCC (TP53, CDKNA2, CASP8, PIK3CA, HRAS, FAT1, TP63, CCND1 and FADD) the analysis identified several candidate novel driver events including mutations of NOTCH3, CSMD3, CRB1, CLTCL1, OSMR and TRPM2, amplification of the proto-oncogenes FOSL1, RELA, TRAF6, MDM2, FRS2 and BAG1, and deletion of the recently described tumor suppressor SMARCC1. Analysis also revealed significantly altered pathways not previously implicated in OSCC including Oncostatin-M signalling pathway, AP-1 and C-MYB transcription networks and endocytosis. There was a trend for higher number of mutations, amplifications and driver events in samples with history of shammah exposure particularly those that tested EBV positive, suggesting an interaction between tobacco exposure and EBV. The work provides further evidence for the genetic heterogeneity of oral cancer and suggests shammah-associated OSCC is characterized by extensive amplification of oncogenes.
Assuntos
Carcinoma de Células Escamosas/epidemiologia , Carcinoma de Células Escamosas/etiologia , Exoma , Sequenciamento de Nucleotídeos em Larga Escala , Neoplasias Bucais/epidemiologia , Neoplasias Bucais/etiologia , Oncogenes , Tabaco sem Fumaça/efeitos adversos , Adulto , Idoso , Biomarcadores , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Mapeamento Cromossômico , Variações do Número de Cópias de DNA , Feminino , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/metabolismo , Neoplasias Bucais/patologia , Mutação , Estadiamento de Neoplasias , Transdução de SinaisRESUMO
Objective: the aim of this study was to evaluate the efficacy of ultrasonography in diagnosis of tempromandibular joint soft tissue injury after mandibular osteosynthesis. Material and Methods: ten male patients (20 joint) with age ranged between 20-28 years were collected from those attending the outpatient clinic of Oral and Maxillofacial Surgery Department, Al-Kuwait Hospital, Sana'a University. Patients were divided into two groups according to the number of fracture line in the mandible. All patients were randomly assigned to diagnosis of the soft tissue changes of temporomandibular joint by either ultrasonography or magnetic resonance image preoperatively, after 2 weeks and 3 months postoperatively. Results: preoperatively, there was moderate agreement between ultrasonography and magnetic resonance image in the diagnosis of abnormal findings in both groups, the difference was not statistically significant. In group I, ultrasonography of the temporomandibular joint didn't detect any abnormal findings after mandibular osteosynthesis, meanwhile, magnetic resonance image recorded abnormal findings 40% and 20% after 2 weeks and 3 months respectively. In group II, the diagnosis of abnormal findings was the same (80%) pre and postoperatively by using magnetic resonance image however, the percent ofabnormal findings was reduced from 60% preoperatively to 40% postoperatively by using ultrasonography. Conclusion: the ultrasonographic image was not able to identify or diagnosis the disc position changes after indirect trauma. However, it had to some extent a role in the identification and diagnosis of effusion in temporomandibular joint. (AU)
Objetivo: o objetivo deste estudo consistiu em avaliar a eficácia da ultrassonografia no diagnóstico de lesões nos tecidos moles da articulação temporomandibular após a osteossíntese mandibular. Material e Métodos: dez pacientes do sexo masculino (no total de 20 indivíduos) de idades entre 20 e 28 anos foram selecionados do serviço ambulatorial do Departamento de Cirurgia Oral e Maxilo-facial, Hospital Al-Kuwait, Universidade de Sana'a. Os indivíduos incluídos foram distribuídos em dois grupos, de acordo com o número de linhas de fratura presentes na mandíbula. Todos os pacientes foram aleatoriamente alocados e divididos, com base no diagnóstico das alterações dos tecidos moles da articulação temporomandibular por ultrassonografia ou ressonância magnética no pré-operatório e em intervalos de 2 semanas e 3 meses no pós-operatório. Resultados: no pré-operatório, houve uma concordância moderada entre a ultrassonografia e a ressonância magnética no diagnóstico de achados anormais em ambos os grupos; a diferença não foi estatisticamente significativa. No grupo I, a ultrassonografia da articulação temporomandibular não revelou quaisquer achados anormais após a osteossíntese mandibular, enquanto a ressonância magnética registou achados anormais em 40% e 20% dos casos após 2 semanas e 3 meses, respectivamente. No grupo II, o diagnóstico das anormalidades por ressonância magnética foi o mesmo (80%) no pré e pós-operatório; contudo, a percentagem de casos anormais por ultrassonografia foi reduzida de 60% no pré-operatório para 40% no pós-operatório. Conclusão: a imagem ultrassonográfica não foi capaz de detectar alterações de posição do disco após trauma indireto. Entretanto, em certa medida, contribuiu para a identificação e diagnóstico de efusão na articulação temporomandibular (AU)
Assuntos
Humanos , Articulação Temporomandibular , Ferimentos e Lesões , Espectroscopia de Ressonância Magnética , Ultrassonografia , Fraturas MandibularesRESUMO
Studies on the possible association between bacteria and oral squamous cell carcinoma (OSCC) remain inconclusive, largely due to methodological variations/limitations. The objective of this study was to characterize the species composition as well as functional potential of the bacteriome associated with OSCC. DNA obtained from 20 fresh OSCC biopsies (cases) and 20 deep-epithelium swabs (matched control subjects) was sequenced for the V1-V3 region using Illumina's 2 × 300 bp chemistry. High quality, non-chimeric merged reads were classified to species level using a prioritized BLASTN-algorithm. Downstream analyses were performed using QIIME, PICRUSt, and LEfSe. Fusobacterium nucleatum subsp. polymorphum was the most significantly overrepresented species in the tumors followed by Pseudomonas aeruginosa and Campylobacter sp. Oral taxon 44, while Streptococcus mitis, Rothia mucilaginosa and Haemophilus parainfluenzae were the most significantly abundant in the controls. Functional prediction showed that genes involved in bacterial mobility, flagellar assembly, bacterial chemotaxis and LPS synthesis were enriched in the tumors while those responsible for DNA repair and combination, purine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, ribosome biogenesis and glycolysis/gluconeogenesis were significantly associated with the controls. This is the first epidemiological evidence for association of F. nucleatum and P. aeruginosa with OSCC. Functionally, an "inflammatory bacteriome" is enriched in OSSC.
Assuntos
Carcinoma de Células Escamosas/etiologia , Infecções por Fusobacterium/complicações , Infecções por Fusobacterium/microbiologia , Fusobacterium nucleatum , Neoplasias Bucais/etiologia , Infecções por Pseudomonas/complicações , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa , Adulto , Idoso , Biodiversidade , Carcinoma de Células Escamosas/patologia , Estudos de Casos e Controles , Feminino , Fusobacterium nucleatum/classificação , Fusobacterium nucleatum/genética , Humanos , Masculino , Metagenoma , Metagenômica/métodos , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/genéticaRESUMO
BACKGROUND: Usefulness of next-generation sequencing (NGS) in assessing bacteria associated with oral squamous cell carcinoma (OSCC) has been undermined by inability to classify reads to the species level. OBJECTIVE: The purpose of this study was to develop a robust algorithm for species-level classification of NGS reads from oral samples and to pilot test it for profiling bacteria within OSCC tissues. METHODS: Bacterial 16S V1-V3 libraries were prepared from three OSCC DNA samples and sequenced using 454's FLX chemistry. High-quality, well-aligned, and non-chimeric reads ≥350 bp were classified using a novel, multi-stage algorithm that involves matching reads to reference sequences in revised versions of the Human Oral Microbiome Database (HOMD), HOMD extended (HOMDEXT), and Greengene Gold (GGG) at alignment coverage and percentage identity ≥98%, followed by assignment to species level based on top hit reference sequences. Priority was given to hits in HOMD, then HOMDEXT and finally GGG. Unmatched reads were subject to operational taxonomic unit analysis. RESULTS: Nearly, 92.8% of the reads were matched to updated-HOMD 13.2, 1.83% to trusted-HOMDEXT, and 1.36% to modified-GGG. Of all matched reads, 99.6% were classified to species level. A total of 228 species-level taxa were identified, representing 11 phyla; the most abundant were Proteobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Actinobacteria. Thirty-five species-level taxa were detected in all samples. On average, Prevotella oris, Neisseria flava, Neisseria flavescens/subflava, Fusobacterium nucleatum ss polymorphum, Aggregatibacter segnis, Streptococcus mitis, and Fusobacterium periodontium were the most abundant. Bacteroides fragilis, a species rarely isolated from the oral cavity, was detected in two samples. CONCLUSION: This multi-stage algorithm maximizes the fraction of reads classified to the species level while ensuring reliable classification by giving priority to the human, oral reference set. Applying the algorithm to OSCC samples revealed high diversity. In addition to oral taxa, a number of human, non-oral taxa were also identified, some of which are rarely detected in the oral cavity.