RESUMO
OBJECTIVE: Reliable biomarkers associated with aggressiveness of non-functioning gonadotroph adenomas (GAs) are lacking. As the growth of tumor remnants is highly variable, molecular markers for growth potential prediction are necessary. We hypothesized that fast- and slow-growing GAs present different gene expression profiles and reliable biomarkers for tumor growth potential could be identified, focusing on the specific role of epithelial-mesenchymal transition (EMT). DESIGN AND METHODS: Eight GAs selected for RNA sequencing were equally divided into fast- and slow-growing group by the tumor volume doubling time (TVDT) median (27.75 months). Data were analyzed by tophat2, cufflinks and cummeRbund pipeline. 40 genes were selected for RT-qPCR validation in 20 GAs based on significance, fold-change and pathway analyses. The effect of silencing MTDH (metadherin) and EMCN (endomucin) on in vitro migration of human adenoma cells was evaluated. RESULTS: 350 genes were significantly differentially expressed (282 genes upregulated and 68 downregulated in the fast group, P-adjusted <0.05). Among 40 selected genes, 11 showed associations with TVDT (-0.669Assuntos
Adenoma/genética
, Transição Epitelial-Mesenquimal/genética
, Neoplasias Hipofisárias/genética
, RNA Mensageiro/metabolismo
, Proteínas Adaptadoras de Transdução de Sinal/genética
, Adenoma/metabolismo
, Adulto
, Idoso
, Caderinas/genética
, Moléculas de Adesão Celular/genética
, Movimento Celular/genética
, Subunidades Catalíticas da Proteína Quinase Dependente de AMP Cíclico/genética
, Feminino
, Hormônio Foliculoestimulante/metabolismo
, Perfilação da Expressão Gênica
, Regulação Neoplásica da Expressão Gênica
, Inativação Gênica
, Humanos
, Técnicas In Vitro
, Peptídeos e Proteínas de Sinalização Intracelular/genética
, Hormônio Luteinizante/metabolismo
, Masculino
, Glicoproteínas de Membrana/genética
, Proteínas de Membrana
, Proteínas Associadas aos Microtúbulos/genética
, Pessoa de Meia-Idade
, Miosina Tipo I/genética
, Proteínas do Tecido Nervoso/genética
, Neoplasias Hipofisárias/metabolismo
, Proteínas Proto-Oncogênicas/genética
, Protocaderinas
, Proteínas de Ligação a RNA
, Receptores de Superfície Celular/genética
, Reação em Cadeia da Polimerase Via Transcriptase Reversa
, Ribonucleases/genética
, Sialoglicoproteínas/genética
, Fatores de Tempo
RESUMO
Sensitive and specific genotyping of human papillomaviruses (HPVs) is important for population-based surveillance of carcinogenic HPV types and for monitoring vaccine effectiveness. Here we compare HPV genotyping by Next Generation Sequencing (NGS) to an established DNA hybridization method. In DNA isolated from urine, the overall analytical sensitivity of NGS was found to be 22% higher than that of hybridization. NGS was also found to be the most specific method and expanded the detection repertoire beyond the 37 types of the DNA hybridization assay. Furthermore, NGS provided an increased resolution by identifying genetic variants of individual HPV types. The same Modified General Primers (MGP)-amplicon was used in both methods. The NGS method is described in detail to facilitate implementation in the clinical microbiology laboratory and includes suggestions for new standards for detection and calling of types and variants with improved resolution.
