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对暴露于X射线或60赫兹磁场的HL60细胞中的基因表达进行BIGEL分析。

BIGEL analysis of gene expression in HL60 cells exposed to X rays or 60 Hz magnetic fields.

作者信息

Balcer-Kubiczek E K, Zhang X F, Han L H, Harrison G H, Davis C C, Zhou X J, Ioffe V, McCready W A, Abraham J M, Meltzer S J

机构信息

Department of Radiation Oncology, School of Medicine; Veterans Affairs Hospital, Baltimore, Maryland 21201, USA.

出版信息

Radiat Res. 1998 Dec;150(6):663-72.

PMID:9840186
Abstract

We screened a panel of 1,920 randomly selected cDNAs to discover genes that are differentially expressed in HL60 cells exposed to 60 Hz magnetic fields (2 mT) or X rays (5 Gy) compared to unexposed cells. Identification of these clones was accomplished using our two-gel cDNA library screening method (BIGEL). Eighteen cDNAs differentially expressed in X-irradiated compared to control HL60 cells were recovered from a panel of 1,920 clones. Differential expression in experimental compared to control cells was confirmed independently by Northern blotting of paired total RNA samples hybridized to each of the 18 clone-specific cDNA probes. DNA sequencing revealed that 15 of the 18 cDNA clones produced matches with the database for genes related to cell growth, protein synthesis, energy metabolism, oxidative stress or apoptosis (including MYC, neuroleukin, copper zinc-dependent superoxide dismutase, TC4 RAS-like protein, peptide elongation factor 1alpha, BNIP3, GATA3, NF45, cytochrome c oxidase II and triosephosphate isomerase mRNAs). In contrast, BIGEL analysis of the same 1,920 cDNAs revealed no differences greater than 1.5-fold in expression levels in magnetic-field compared to sham-exposed cells. Magnetic-field-exposed and control samples were analyzed further for the presence of mRNA encoding X-ray-responsive genes by hybridization of the 18 specific cDNA probes to RNA from exposed and control HL60 cells. Our results suggest that differential gene expression is induced in approximately 1% of a random pool of cDNAs by ionizing radiation but not by 60 Hz magnetic fields under the present experimental conditions.

摘要

我们筛选了一组1920个随机选择的cDNA,以发现与未暴露细胞相比,在暴露于60Hz磁场(2mT)或X射线(5Gy)的HL60细胞中差异表达的基因。使用我们的双凝胶cDNA文库筛选方法(BIGEL)完成了这些克隆的鉴定。从1920个克隆中回收了18个与对照HL60细胞相比在X射线照射下差异表达的cDNA。通过与18个克隆特异性cDNA探针中的每一个杂交的配对总RNA样品的Northern印迹,独立证实了与对照细胞相比实验细胞中的差异表达。DNA测序显示,18个cDNA克隆中的15个与细胞生长、蛋白质合成、能量代谢、氧化应激或凋亡相关基因的数据库匹配(包括MYC、神经白细胞素、铜锌依赖性超氧化物歧化酶、TC4 RAS样蛋白、肽延伸因子1α、BNIP3、GATA3、NF45、细胞色素c氧化酶II和磷酸丙糖异构酶mRNA)。相比之下,对相同的1920个cDNA进行的BIGEL分析显示,与假暴露细胞相比,磁场暴露细胞的表达水平差异不超过1.5倍。通过将18个特异性cDNA探针与暴露和对照HL60细胞的RNA杂交,进一步分析磁场暴露和对照样品中编码X射线反应基因的mRNA的存在情况。我们的结果表明,在目前的实验条件下,电离辐射可诱导约1%的随机cDNA文库中出现差异基因表达,而60Hz磁场则不会。

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