DSM-Department of Medical Sciences, University of Trieste, 34149 Trieste, Italy.
DAME-Department of Medical Area, University of Udine, 33100 Udine, Italy.
Int J Mol Sci. 2023 Jun 29;24(13):10836. doi: 10.3390/ijms241310836.
In forensic medicine, identifying novel biomarkers for use as diagnostic tools to ascertain causes of death is challenging because of sample degradation. To that aim, a cohort ( = 26) of fatal traumatic brain injuries (TBIs) were tested for three candidate miRNAs (namely, miR-124-3p, miR-138-5p, and miR144-3p). For each case, three FFPE specimens (coup area (CA), contrecoup area (CCA), and the corpus callosum (CC)) were investigated, whereas the FFPE brain tissues of 45 subjects (deceased due to acute cardiovascular events) were used as controls. Relative quantification via the ∆∆Ct method returned significantly higher expression levels of the three candidate miRNAs ( < 0.01) in the TBI cases. No difference was detected in the expression levels of any miRNA investigated in the study among the CA, CCA, and CC. Furthermore, the analyzed miRNAs were unrelated to the TBI samples' post-mortem intervals (PMIs). On the contrary, ahas were significantly correlated ( < 0.01) with the agonal time in TBI deaths. Since the RNA was highly degraded in autoptic FFPE tissues, it was impossible to analyze the mRNA targets of the miRNAs investigated in the present study, highlighting the necessity of standardizing pre-analytical processes even for autopsy tissues.
在法医学中,由于样本降解,鉴定可用于确定死因的新型生物标志物作为诊断工具具有挑战性。为此,对 26 例致命性创伤性脑损伤 (TBI) 患者进行了三种候选 miRNA(即 miR-124-3p、miR-138-5p 和 miR144-3p)的检测。对于每个病例,均检测了三个 FFPE 标本(挫伤区 (CA)、对冲区 (CCA) 和胼胝体 (CC)),而 45 例因急性心血管事件死亡的 FFPE 脑组织则作为对照。通过 ∆∆Ct 方法进行的相对定量分析显示,三种候选 miRNA 的表达水平明显升高(<0.01)。在 CA、CCA 和 CC 中,未检测到研究中任何 miRNA 的表达水平存在差异。此外,分析的 miRNA 与 TBI 样本的死后间隔时间 (PMI) 无关。相反,ahas 与 TBI 死亡患者的濒死时间呈显著相关(<0.01)。由于在尸检 FFPE 组织中 RNA 高度降解,因此无法分析本研究中检测到的 miRNA 的 mRNA 靶标,这突出了即使对于尸检组织也需要标准化分析前过程的必要性。