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 基因的体细胞突变。因此PIG-A基因突变被认为是PNH发生的原因。然而，最近的研究表明，健康个体中也存在罕见的 PNH 细胞（Araten，DJ 等人，1996）。因此，推测除了PIG-A基因突变之外，PNH克隆的生长优势和/或生存优势对于PNH的发展也是不可或缺的。本研究的目的是阐明与 PNH 克隆的生长优势和/或生存优势相关的基因。我们进行了 cDNA 消减和 cDNA 宏阵列，以筛选与 PNH 患者的 CD59^+ 粒细胞相比，PNH 患者的 CD59^- 粒细胞中上调的基因。通过半定量 RT-PCR 检测，一名 PNH 患者的 CD59^- 粒细胞中的 3 个基因上调超过 2 倍。然而，与其他 3 名 PNH 患者的 CD59^+ 粒细胞相比，我们没有发现这些基因在 CD59^- 粒细胞中上调的显着趋势。此前已有报道称 PNH 患者中有 4 个基因表达上调。我们通过半定量 RT-PCR 检查了 7 名 PNH 患者和 5 名健康供体中这 4 个基因的表达。我们在 PNH 患者中没有发现这 4 个基因上调的趋势。