SCREENING FOR THE GENES RESPONSIBLE FOR THE SURVIVAL ADVANTAGE OF PNH CLONES

筛选负责 PNH 克隆生存优势的基因

• 批准号: 12671012
• 负责人: KANAMARU Akihisa
• 金额: $ 2.18万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671012/
• 关键词: PNH; PNH clone; cDNA subtraction; survival advantage; cloning; PIG-A mutation; SMART-PCR; cDNAサブトラクション; SMART-PCR法; cDNAマクロアレイ

Somatic mutations in the PIG-A gene have been found in all PNH cases. Therefore PIG-A gene mutation has been considered to be responsible for the development of PNH. However, recent study revealed that rare PNH cells are present also in healthy individuals (Araten, DJ et al. 1996). Therefore, it is postulated that growth advantage and/or survival advantage of PNH clones is indispensable for the development of PNH, in addition to PIG-A gene mutation. The purpose of this study is to clarify the genes concerned in growth advantage and/or survival advantage of PNH clones. We performed a cDNA subtraction and a cDNA macro array to screen for the genes up-regulated in CD59^- granulocytes of PNH patient compared to CD59^+ granulocytes of PNH patient. Three genes were up-regulated more than 2 times in CD59^- granulocytes of one PNH patient by semi-quantitative RT-PCR assay. However we could not find remarkable tendency that these genes are up-regulated in CD59^- granulocytes compared to CD59^+ granulocytes in 3 other PNH patients. Already, 4 genes were reported up-regulated in PNH patients before. We have checked the expression of these 4 genes in 7 PNH patients and in 5 healthy donors by semi-quantitative RT-PCR. We could not find tendency that these 4 genes were up-regulated in our PNH patients.

在所有PNH病例中都发现了PIG-A基因中的体细胞突变。因此，PIG-A基因突变被认为是PNH发展的原因。然而，最近的研究表明，健康个体也存在罕见的PNH细胞（Araten，DJ等，1996）。因此，据推测，除PIG-A基因突变外，PNH克隆的生长优势和/或生存优势对于PNH的发展是必不可少的。这项研究的目的是阐明PNH克隆的生长优势和/或生存优势有关的基因。与PNH患者的CD59^+粒细胞相比，我们对PNH患者的CD59^ - 粒细胞中上调的基因进行了筛选cDNA减法和cDNA宏阵列。通过半定量RT-PCR分析，在一名PNH患者的CD59^ - 粒细胞中，三个基因上调了2倍以上。但是，与CD59^ - 粒细胞相比，与其他3名PNH患者的CD59^+粒细胞相比，我们找不到这些基因在CD59^ - 粒细胞中上调的趋势。以前，PNH患者已经上调了4个基因。我们已经通过半定量RT-PCR检查了7名PNH患者和5个健康供体中这4个基因的表达。我们找不到在PNH患者中上调这4个基因的趋势。