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Limitations of high throughput methods for miRNA expression profiles in non-functioning pituitary adenomas.
Darvasi, O; Szabo, P M; Nemeth, K; Szabo, K; Spisak, S; Liko, I; Czirjak, S; Racz, K; Igaz, P; Patocs, A; Butz, Henriett.
Afiliación
  • Darvasi O; Hereditary Endocrine Tumors Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Szabo PM; Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Nemeth K; 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
  • Szabo K; 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
  • Spisak S; Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Liko I; Hereditary Endocrine Tumors Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Czirjak S; National Institute of Neurosurgery, Budapest, Hungary.
  • Racz K; Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Igaz P; 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
  • Patocs A; Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary.
  • Butz H; 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
Pathol Oncol Res ; 25(1): 169-182, 2019 Jan.
Article en En | MEDLINE | ID: mdl-29043608
Microarray, RT-qPCR based arrays and next-generation-sequencing (NGS) are available high-throughput methods for miRNA profiling (miRNome). Analytical and biological performance of these methods were tested in identification of biologically relevant miRNAs in non-functioning pituitary adenomas (NFPA). miRNome of 4 normal pituitary (NP) and 8 NFPA samples was determined by these platforms and expression of 21 individual miRNAs was measured on 30 (20 NFPA and 10 NP) independent samples. Complex bioinformatics was used. 132 and 137 miRNAs were detected by all three platforms in NP and NFPA, respectively, of which 25 were differentially expressed (fold change > 2). The strongest correlation was observed between microarray and TaqMan-array, while the data obtained by NGS were the most discordant despite of various bioinformatics settings. As a technical validation we measured the expression of 21 selected miRNAs by individual RT-qPCR and we were able to validate 35.1%, 76.2% and 71.4% of the miRNAs revealed by SOLiD, TLDA and microarray result, respectively. We performed biological validation using an extended number of samples (20 NFPAs and 8 NPs). Technical and biological validation showed high correlation (p < 0.001; R = 0.96). Pathway and network analysis revealed several common pathways but no pathway showed the same activation score. Using the 25 platform-independent miRNAs developmental pathways were the top functional categories relevant for NFPA genesis. The difference among high-throughput platforms is of great importance and selection of screening method can influence experimental results. Validation by another platform is essential in order to avoid or to minimalize the platform specific errors.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neoplasias Hipofisarias / Biomarcadores de Tumor / Regulación Neoplásica de la Expresión Génica / Perfilación de la Expresión Génica / MicroARNs Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Pathol Oncol Res Asunto de la revista: NEOPLASIAS / PATOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Suiza
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neoplasias Hipofisarias / Biomarcadores de Tumor / Regulación Neoplásica de la Expresión Génica / Perfilación de la Expresión Génica / MicroARNs Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Pathol Oncol Res Asunto de la revista: NEOPLASIAS / PATOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Hungria Pais de publicación: Suiza