IDENTIFICATION OF THE CAUSATIVE GENES RESPONSIBLE FOR ERYTHROID APOPTOSIS INDUCED BY GLYCOLYTIC INHIBITION

糖酵解抑制诱导的红细胞凋亡的致病基因的鉴定

• 批准号: 16590254
• 负责人: KANNO HITOSHI
• 金额: $ 2.24万
• 依托单位: TOKYO WOMEN'S MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590254/
• 关键词: hemolytic anemia; inborn error of metabolism; red cell life span; anti-oxidant; 赤血球; 無効造血; 変異マウス; 造血細胞コロニー解析; ピルビン酸キナーゼ; 赤血球分化; 酸化ストレス

We previously established a Friend erythroleukemic cell, SLC3, from the mice model of red blood cell pyruvate kinase (R-PK) deficiency. SLC3 showed spontaneous apoptosis during routine passage. When cultured in a condition of glucose deprivation or supplementation with 2-deoxyglucose, a control Friend cell, CBA2, also showed apoptosis. Preincubation with N-acetyl cysteine, glutathione precursor, reduces apoptosis of CBA2 induced by 2-deoxyglucose, suggesting that glycolytic inhibition increases oxidative stress in erythroid cells and that erythroid apoptosis is likely to be induced by reactive oxygen species (ROS).When cultured in 5mM phosphoenolpyruvate (PEP) for 48 hours, intracellular concentration of PEP and pyruvate increased up to 7- and 2-times, respectively. ROS in the PEP-treated erythroid cells significantly decreased, and apoptotic cell number decreased about 50% of non-treated cells. This observation suggests that accumulation of PEP improves glycolysis since the mutant R-PK has lower substrate specificity due to the active site mutation. Taken together, we conclude that glycolytic inhibition increases oxidative stress in erythroid cells and activate proapoptotic gene expressions, leading to apoptosis.

我们以前建立了一个朋友红白血病细胞，SLC 3，从小鼠模型的红细胞丙酮酸激酶（R-PK）缺陷。SLC 3在常规传代过程中显示自发凋亡。当在葡萄糖剥夺或补充2-脱氧葡萄糖的条件下培养时，对照Friend细胞CBA 2也显示凋亡。用谷胱甘肽前体N-乙酰半胱氨酸预孵育CBA 2，可减少2-脱氧葡萄糖诱导的CBA 2凋亡，表明糖酵解抑制增加了红系细胞的氧化应激，红系细胞凋亡可能是由活性氧（ROS）诱导的。PEP处理的红系细胞中的ROS显著减少，凋亡细胞数减少约50%的未处理的细胞。这一观察结果表明PEP的积累改善了糖酵解，因为突变体R-PK由于活性位点突变而具有较低的底物特异性。综上所述，我们得出结论，糖酵解抑制增加了红系细胞的氧化应激，激活了促凋亡基因的表达，导致细胞凋亡。

项目成果

Histopathological study of lattice corneal dystrophy with L527R mutation of transforming growth factor-beta induced gene.

转化生长因子-β诱导基因L527R突变引起的格子状角膜营养不良的组织病理学研究。

• 发表时间: 2004
• 期刊: Nippon Ganka Gakkai Zasshi 108
• 作者: Nakagawa E;et al.
• 通讯作者: et al.

Cyclic polylactate inhibited growth of cloned leukemic cells through reducing glycolytic enzyme activities.

环状聚乳酸通过降低糖酵解酶活性来抑制克隆白血病细胞的生长。

• 发表时间: 2005
• 期刊: Oncology Report 14
• 作者: Harada Y;et al.
• 通讯作者: et al.

Ineffective erythropoiesis in mutant mice with deficient pyruvate kinase activity

• DOI: 10.1016/j.exphem.2005.07.008
• 发表时间: 2005-11-01
• 期刊: EXPERIMENTAL HEMATOLOGY
• 影响因子: 2.6
• 作者: Aizawa, S;Harada, T;Fujii, H
• 通讯作者: Fujii, H

Molecular basis of Japanese variants of pyrimidine 5′-nucleotidase deficiency

• DOI: 10.1111/j.1365-2141.2004.05029.x
• 发表时间: 2004-07-01
• 期刊: BRITISH JOURNAL OF HAEMATOLOGY
• 影响因子: 6.5
• 作者: Kanno, H;Takizawa, T;Fujii, H
• 通讯作者: Fujii, H

A novel homozygous mutation of PKLR gene in a pyruvate-kinase-deficient Korean family

• DOI: 10.1159/000084453
• 发表时间: 2005-01-01
• 期刊: ACTA HAEMATOLOGICA
• 影响因子: 2.4
• 作者: Park-Hah, JO;Kanno, H;Fujii, H
• 通讯作者: Fujii, H