APOPTOSIS OF HUMAN ERYTHROID COLONY-FORMING CELLS IS DECREASED BY STEM-CELL FACTOR AND INSULIN-LIKE GROWTH FACTOR-I AS WELL AS ERYTHROPOIETIN
APOPTOSIS OF HUMAN ERYTHROID COLONY-FORMING CELLS IS DECREASED BY STEM-CELL FACTOR AND INSULIN-LIKE GROWTH FACTOR-I AS WELL AS ERYTHROPOIETIN
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DOI：
10.1002/jcp.1041560207
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发表时间：
1993-08-01
影响因子：
5.6
通讯作者：
KRANTZ, SB
中科院分区：
文献类型：
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作者：
MUTA, K;KRANTZ, SB
为了阐明促红细胞生成素（EP）、干细胞因子（SCF）或胰岛素样生长因子I（IGF-I）调节红细胞生成的方式，研究了人红系祖细胞的凋亡。从人外周血中部分纯化出红系爆发形成单位（BFU-E），经6天培养，获得以红系集落形成单位（CFU-E）为主的红系集落形成细胞（ECFC）。用[H-3]胸苷标记细胞，在无血清液体培养基中于37 ℃孵育16 h，通过琼脂糖凝胶电泳分析DNA断裂模式。当这些细胞在没有EP的情况下孵育时，70%的总细胞DNA被分解成小于5千碱基的DNA片段，并且细胞核浓缩和碎片（细胞凋亡的特征）明显。当EP将DNA分解的量大大减少到23%时，SCF和IGF-I各自将DNA分解的量减少到38-46%，并且当加在一起时，减少到24%。SCF和IGF-I的剂量反应实验表明，在无血清半固体培养物中刺激集落形成的浓度下，DNA片段呈剂量依赖性减少。最后，在无血清液体培养后，在37 ℃下培养16小时，通过血浆凝块法进行的ECFC测定表明，在不存在EP的情况下，SCF或IGF-I以及EP本身对红系细胞集落形成能力具有显著的保护作用。这些数据表明，人红系祖细胞发生细胞凋亡，这是减少SCF和IGF-I以及EP和这些因素的控制细胞凋亡的每一个有一个突出的作用，在红细胞生成的调节。(C)1993 Wiley-Liss，Inc.
To clarify the manner by which erythropoietin (EP), stem cell factor (SCF), or insulin-like growth factor I (IGF-I) regulate erythropoiesis, apoptosis of human erythroid progenitor cells was investigated. Human burst-forming units-erythroid (BFU-E) partially purified from peripheral blood were cultured for 6 days to generate erythroid colony-forming cells (ECFC), which consist mainly of colony-forming units-erythroid (CFU-E). The cells were labeled with [H-3]thymidine, incubated in serum-free liquid media, at 37-degrees-C, for 16 h, and the pattern of DNA breakdown was analyzed by agarose gel electrophoresis. When these cells were incubated without EP, 70% of the total cellular DNA was broken down into DNA fragments of less than 5 kilobases and nuclear condensation and fragmentation, characteristic of apoptosis, were evident. While EP greatly reduced the amount of DNA breakdown to 23%, SCF and IGF-I each reduced the amount of DNA breakdown to 38-46% and, when added together, to 24%. Dose-response experiments with SCF and IGF-I showed a dose-dependent reduction in DNA fragmentation at concentrations that stimulate colony formation in serum-free semisolid cultures. Finally, assays of ECFC performed by the plasma clot method, after serum-free liquid culture, at 37-degrees-C, for 16 h, demonstrated marked protection of erythroid colony-forming capacity by SCF or IGF-I in the absence of EP, as well as by EP itself. These data indicate that human erythroid progenitor cells undergo apoptosis which is reduced by SCF and IGF-I as well as EP and suggest that the control of apoptosis by each of these factors has a prominent role in the regulation of erythropoiesis. (C) 1993 Wiley-Liss, Inc.