Corbin Monique V, Rockx Davy A P, Oostra Anneke B, Joenje Hans, Dorsman Josephine C
Department of Clinical Genetics, Section Oncogenetics, VU University Medical Center, 1081BT Amsterdam, The Netherlands.
Free Radic Biol Med. 2015 Dec;89:863-72. doi: 10.1016/j.freeradbiomed.2015.08.026. Epub 2015 Oct 9.
Aim of this study was to explore cellular changes associated with increased resistance to atmospheric oxygen using high-resolution DNA and RNA profiling combined with functional studies. Two independently selected oxygen-resistant substrains of HeLa cells (capable of proliferating at >80% O2, i.e. hyperoxia) were compared with their parental cells (adapted to growth at 20% O2, but unable to grow at >80% O2). A striking consistent alteration found to be associated with the oxygen-resistant state appeared to be an amplified and overexpressed region on chromosome 16p13.3 harboring 21 genes. The driver gene of this amplification was identified by functional studies as NARFL, which encodes a component of the cytosolic iron-sulfur cluster assembly system. In line with this result we found the cytosolic c-aconitase activity as well as the nuclear protein RTEL1, both Fe-S dependent proteins, to be protected by NARFL overexpression under hyperoxia. In addition, we observed a protective effect of NARFL against hyperoxia-induced loss of sister-chromatid cohesion. NARFL thus appeared to be a key factor in the cellular defense against hyperoxia-induced oxidative stress in human cells. Our findings suggest that new insight into age-related degenerative processes may come from studies that specifically address the involvement of iron-sulfur proteins.
本研究的目的是利用高分辨率DNA和RNA分析结合功能研究,探索与大气氧抗性增加相关的细胞变化。将两个独立选择的耐氧HeLa细胞亚系(能够在>80%氧气条件下增殖,即高氧环境)与其亲代细胞(适应于在20%氧气条件下生长,但在>80%氧气条件下无法生长)进行比较。发现与耐氧状态相关的一个显著一致的变化似乎是16号染色体p13.3上一个包含21个基因的区域发生了扩增和过表达。通过功能研究确定该扩增的驱动基因是NARFL,它编码胞质铁硫簇组装系统的一个组分。与这一结果一致,我们发现在高氧条件下,胞质顺乌头酸酶活性以及核蛋白RTEL1(这两种都是铁硫依赖性蛋白)受到NARFL过表达的保护。此外,我们观察到NARFL对高氧诱导的姐妹染色单体粘连丧失具有保护作用。因此,NARFL似乎是人类细胞抵御高氧诱导氧化应激的关键因素。我们的研究结果表明,对与年龄相关的退行性过程的新认识可能来自于专门研究铁硫蛋白参与情况的研究。