We aimed to identify novel markers for aggressive prostate cancer in a STAT3-low proteomics-derived dataset of mitochondrial proteins by immunohistochemical analysis and correlation with transcriptomic data and biochemical recurrence in a STAT3 independent PCa cohort. Formalin-fixed paraffin-embedded tissue (FFPE) sample selection for proteomic analysis and tissue-microarray (TMA) generation was conducted from a cohort of PCa patients. Retrospective data analysis was performed with the same cohort. 153 proteins differentially expressed between STAT3-low and STAT3-high samples were identified. Out of these, 46 proteins were associated with mitochondrial processes including oxidative phosphorylation (OXPHOS), and 45 proteins were upregulated, including NDUFS1/ATP5O. In a STAT3 independent PCa cohort, high expression of NDUFS1/ATP5O was confirmed by immunocytochemistry (IHC) and was significantly associated with earlier biochemical recurrence (BCR). mRNA expression levels for these two genes were significantly higher in intra-epithelial neoplasia and in PCa compared to benign prostate glands. NDUFS1/ATP5O levels are increased both at the mRNA and protein level in aggressive PCa. Our results provide evidence that NDUFS1/ATP5O could be used to identify high-risk PCa patients.

• Publikační typ
• časopisecké články MeSH

Early neonatal adaptation to extrauterine life is i.a. dependent on effective mitochondrial biogenesis during foetal development. Understanding of mitochondrial biogenesis is limited, because only scarce data are available from prenatal studies including RNA analyses in human foetal tissues. Aims of the study were focused on the factors affecting RNA quality in human placental tissue (HPT) including temperature, time period before HPT freezing and the Apgar score. In addition, optimal reference genes for mRNA quantification by real-time PCR in HPT were studied. Samples of HPT were obtained after the birth of 20 term neonates. Seven HPT were used for the time-course study of RNA degradation in two different temperatures (0 °C and 24 °C). Various instruments NanoDrop (NanoDrop Technologies), Experion (Bio-Rad Laboratories), Agilent 2100 Bioanalyzer (Agilent Technologies) were used for analysis of RNA integrity, purity and yield. Identification of suitable reference genes was achieved by analysing six candidate genes (ATP5O, SDHA, TBP, HPRT, PMBS, ATP6) for their expression stability (GeNorm application). The results showed that the HPT samples for RNA analyses must be frozen immediately after birth in –80 °C or stored at 0 °C maximally for 1 hour. The reference genes ATP50 and SDHA were the most stable for mRNA quantification in HPT. Human placenta represents easily obtainable source of foetal tissue for studies concerning mitochondrial biogenesis. We demonstrated that the critical limit for optimal storage and handling of HPT are the temperature and the time period before freezing of the samples.

• MeSH
• Apgar skóre MeSH
• financování organizované MeSH
• lidé MeSH
• novorozenec MeSH
• placenta chemie   MeSH
• plod MeSH
• polymerázová řetězová reakce MeSH
• RNA analýza   MeSH
• těhotenství MeSH
• teplota MeSH
• uchovávání tkání MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH