Trinity Translational Medicine Institute, Department of Surgery, St. James's Hospital, D08W9RT Dublin, Ireland.
Department of Clinical Medicine, Trinity Translational Medicine Institute, Trinity College Dublin and St. James's Hospital, D08W9RT Dublin, Ireland.
Int J Mol Sci. 2018 Nov 6;19(11):3483. doi: 10.3390/ijms19113483.
Barrett's esophagus and esophageal cancer lack prognostic markers that allow the tailoring of personalized medicine and biomarkers with potential to provide insight into treatment response. This study aims to characterize mitochondrial function across the metaplasia-dysplasia-adenocarcinoma disease sequence in Barrett's esophagus and examines the functional effect of manipulating mitochondrial genes. Mitochondrial genes of interest were validated in in vitro cell lines across the metaplasia (QH), dysplasia (GO) and adenocarcinoma (OE33) sequence and in in vivo patient tissue samples. These genes were subsequently knocked down in QH and OE33 cells and the functional effect of siRNA-induced knockdown on reactive oxygen species production, mitochondrial mass, mitochondrial membrane potential and cellular metabolism was investigated. Three global mitochondrial genes (, and ) were differentially altered across the in vivo Barrett's disease sequence. We also demonstrate that knockdown of , and in vitro resulted in significant alterations in mitochondrial membrane potential; however, no differences in reactive oxygen species or mitochondrial mass were observed. Furthermore, knockdown of these genes in esophageal adenocarcinoma cells significantly altered cellular metabolism. In conclusion, we found that differential expression of , , and were altered across the Barrett's disease sequence and manipulation of these genes elicited significant effects on mitochondrial membrane potential.
巴雷特食管和食管癌缺乏预后标志物，无法实现个体化医学的精准定制，也缺乏有潜力用于预测治疗反应的生物标志物。本研究旨在描述巴雷特食管的化生-异型增生-腺癌疾病序列中的线粒体功能，并探讨操纵线粒体基因的功能效应。在体外细胞系中验证了感兴趣的线粒体基因在化生（QH）、异型增生（GO）和腺癌（OE33）序列中的功能，以及在体内患者组织样本中的功能。随后在 QH 和 OE33 细胞中敲低这些基因，并研究 siRNA 诱导的敲低对活性氧产生、线粒体质量、线粒体膜电位和细胞代谢的功能影响。三个全局线粒体基因（、和）在体内巴雷特病序列中差异改变。我们还证明，体外敲低这些基因导致线粒体膜电位显著改变；然而，活性氧或线粒体质量没有观察到差异。此外，这些基因在食管腺癌细胞中的敲低显著改变了细胞代谢。总之，我们发现、和的差异表达在巴雷特病序列中发生改变，并且这些基因的操纵对线粒体膜电位产生显著影响。
