Notch1-induced delay of human hematopoietic progenitor cell differentiation is associated with altered cell cycle kinetics
Notch1-induced delay of human hematopoietic progenitor cell differentiation is associated with altered cell cycle kinetics
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DOI：
10.1182/blood.v93.3.838.403k29_838_848
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发表时间：
1999-02-01
期刊：
影响因子：
20.3
通讯作者：
Scadden, DT
中科院分区：
文献类型：
--
作者：
Carlesso, N;Aster, JC;Scadden, DT
造血是增殖和分化之间的平衡，可能受到环境信号的调节，Notch受体及其配体在进化过程中高度保守，并已被证明在多个发育系统中调节细胞命运决定。为了评估Notch 1信号传导是否可以调节人类造血以维持细胞处于未成熟状态，我们将基于水泡性口炎病毒G蛋白（VSV-G）的假型双顺反子鼠干细胞病毒（MSCV）的逆转录病毒载体转导到分化能力HL-60细胞系和原代脐带血来源的CD 34（+）细胞中，所述逆转录病毒载体表达Notch 1（ICN）的组成型活性形式和绿色荧光蛋白。此外，我们观察到HL-60细胞和原代CD 34（+）细胞表面上的内源性Notch 1表达，因此将细胞暴露于NIH 3 T3细胞上表达的Notch配体Jagged 2。Notch的配体非依赖性和配体依赖性激活均导致HL-60细胞和脐血CD 34（+）细胞获得分化标志物的延迟。此外，原代CD 34（+）细胞保留了形成未成熟集落的能力，即集落形成单位混合物（CFU-mix），而对照细胞失去了这种能力。Notch 1的激活与HL-60细胞在获得成熟细胞表型之前处于G（0）/G（1）期的比例减少相关。尽管保持了细胞的增殖率和细胞周期的总长度，但仍注意到这种通过G（1）的增强进展。这些发现表明，Notch 1激活延迟人造血分化，并表明Notch分化效应与细胞周期动力学改变有关。(C)1999年，美国血液学会。
Hematopoiesis is a balance between proliferation and differentiation that may be modulated by environmental signals, Notch receptors and their ligands are highly conserved during evolution and have been shown to regulate cell fate decisions in multiple developmental systems. To assess whether Notch1 signaling may regulate human hematopoiesis to maintain cells in an immature state, we transduced a vesicular stomatitis virus G-protein (VSV-G) pseudo-typed bicistronic murine stem cell virus (MSCV)-based retroviral vector expressing a constitutively active form of Notch1 (ICN) and green fluorescence protein into the differentiation competent HL-60 cell line and primary cord blood-derived CD34(+) cells. In addition, we observed endogenous Notch1 expression on the surface of both HL-60 cells and primary CD34(+) cells, and therefore exposed cells to Notch ligand Jagged2, expressed on NIH3T3 cells. Both ligand-independent and ligand-dependent activation of Notch resulted in delayed acquisition of differentiation markers by HL-60 cells and cord blood CD34(+) cells. In addition, primary CD34(+) cells retained their ability to form immature colonies, colony-forming unit-mix (CFU-mix), whereas control cells lost this capacity. Activation of Notch1 correlated with a decrease in the fraction of HL-60 cells that were in G(0)/G(1) phase before acquisition of a mature cell phenotype. This enhanced progression through G(1) was noted despite preservation of the proliferative rate of the cells and the overall length of the cell cycle. These findings show that Notch1 activation delays human hematopoietic differentiation and suggest a link of Notch differentiation effects with altered cell cycle kinetics. (C) 1999 by The American Society of Hematology.