Kabanova Sveta, Kleinbongard Petra, Volkmer Jens, Andrée Birgit, Kelm Malte, Jax Thomas W
Department of Medicine, Universitätsklinikum Düsseldorf, Heinrich-Heine-University, Germany.
Int J Med Sci. 2009;6(4):156-9. doi: 10.7150/ijms.6.156. Epub 2009 Apr 28.
Understanding of molecular mechanisms governing the enucleating phenomena of human erythrocytes is of major importance in both fundamental and applied studies. Total RNA (n=7) from human RBCs (purity of erythrocyte preparation >99,99%) was tested using 2100 Bioanalyzer (Agilent, USA), and transcribed to cDNA. Microarray analysis was performed with the Human Genome Focus GeneChip (Affymetrix, USA), containing 8500 transcripts corresponding to 8400 human genes. Here we report that human RBCs contain typical eukaryotic RNA with 28S- and18S-rRNA standard bands. Microarray studies revealed the presence of transcripts of 1019 different genes in erythrocytic RNA. Gene Ontology analysis recognized 859 genes involved in general biological processes: 529 genes for cellular metabolism, 228 genes for signal transduction, 104 genes for development, 107 genes for immune response, 62 genes for protein localization, 53 genes for programmed cell death, and 5 genes for autophagy. A number of genes responsible for transcription, translation, RNA-stabilisation as well as for apoptosis and anti-apoptosis have been identified for the first time in circulating human RBCs. The presented data shed new light on the genetic determination of erythropoiesis, apoptosis and may have implications on the pathophysiology and diagnosis of various diseases involving red blood cells.
了解控制人类红细胞去核现象的分子机制在基础研究和应用研究中都具有重要意义。使用2100生物分析仪(美国安捷伦公司)对来自人类红细胞的总RNA(n = 7)(红细胞制备纯度>99.99%)进行检测,并转录为cDNA。使用包含对应于8400个人类基因的8500个转录本的人类基因组聚焦基因芯片(美国Affymetrix公司)进行微阵列分析。在此我们报告,人类红细胞含有具有28S和18S - rRNA标准条带的典型真核RNA。微阵列研究揭示了红细胞RNA中存在1019个不同基因的转录本。基因本体分析识别出859个参与一般生物学过程的基因:529个参与细胞代谢的基因、228个参与信号转导的基因、104个参与发育的基因、107个参与免疫反应的基因、62个参与蛋白质定位的基因、53个参与程序性细胞死亡的基因以及5个参与自噬的基因。首次在循环中的人类红细胞中鉴定出许多负责转录、翻译、RNA稳定以及凋亡和抗凋亡的基因。所呈现的数据为红细胞生成、凋亡的遗传决定提供了新的线索,并且可能对涉及红细胞的各种疾病的病理生理学和诊断产生影响。