Analysis of the biologic functions of H- and L-ferritins in HeLa cells by transfection with siRNAs and cDNAs: evidence for a proliferative role of L-ferritin
Analysis of the biologic functions of H- and L-ferritins in HeLa cells by transfection with siRNAs and cDNAs: evidence for a proliferative role of L-ferritin
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DOI：
10.1182/blood-2003-06-1842
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发表时间：
2004-03-15
期刊：
影响因子：
20.3
通讯作者：
Arosio, P
中科院分区：
文献类型：
--
作者：
Cozzi, A;Corsi, B;Arosio, P
我们描述了使用小干扰 RNA (siRNA) 下调 HeLa 细胞中的 H 和 L 铁蛋白水平。在稳定转染和瞬时转染中，siRNA 将 H 铁蛋白和 L 铁蛋白表达抑制至背景水平的约 20% 至 25%。平行分析用 H-和 L-铁蛋白 cDNA 转染的 HeLa 细胞，以比较铁蛋白上调和下调的效果。我们发现，L-铁蛋白水平的大幅改变并不影响HeLa细胞中铁的利用率，而是以不依赖于铁的方式对细胞增殖率产生积极影响。正如预期的那样，H-铁蛋白的瞬时下调改变了细胞铁的利用率和对氧化损伤的抵抗力。相比之下，转染 siRNA 的 HeLa 细胞克隆中 H-铁蛋白的稳定抑制并没有增加细胞铁的利用率，而是使细胞对铁补充和螯合的抵抗力降低。结果表明，L-铁蛋白对 HeLa 细胞中的细胞铁稳态没有直接影响，但它具有与铁无关的新功能。此外，他们认为 H-铁蛋白的功能是充当铁缓冲剂。 (C) 2004 年，美国血液学会。
We describe the use of small interfering RNAs (siRNAs) to down-regulate H- and L-ferritin levels in HeLa cells. siRNAs repressed H- and L-ferritin expression to about 20% to 25% of the background level in both stable and transient transfections. HeLa cells transfected with H- and L-ferritin cDNAs were analyzed in parallel to compare the effects of ferritin up- and down-regulation. We found that large modifications of L-ferritin levels did not affect iron availability in HeLa cells but positively affected cell proliferation rate in an iron-independent manner. The transient down-regulation of H-ferritin modified cellular iron availability and resistance to oxidative damage, as expected. In contrast, the stable suppression of H-ferritin in HeLa cell clones transfected with siRNAs did not increase cellular iron availability but made cells less resistant to iron supplementation and chelation. The results indicate that L-ferritin has no direct effects on cellular iron homeostasis in HeLa cells, while it has new, iron-unrelated functions. In addition, they suggest that H-ferritin function is to act as an iron buffer. (C) 2004 by The American Society of Hematology.