Stage-related proliferative activity determines c-myb functional requirements during normal human hematopoiesis.
Stage-related proliferative activity determines c-myb functional requirements during normal human hematopoiesis.
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阶段相关的增殖活性决定了正常人类造血过程中 c-myb 的功能需求。
DOI：
10.1172/jci114433
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Calabretta,B
中科院分区：
文献类型：
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作者：
Caracciolo,D;Venturelli,D;Valtieri,M;Peschle,C;Gewirtz,AM;Calabretta,B
为了确定在正常人类造血的特定阶段是否优先需要MYB蛋白，我们将正常骨髓单个核细胞(MNC)与c-MYB反义寡核苷酸孵育。处理后的细胞在半固体培养条件下培养，以利于特定的祖细胞类型的生长。与未经处理的对照组相比，重组人(Rh)IL-3诱导的粒-巨噬细胞(GM)CFU集落减少77%，rh GM集落刺激因子(CSF)刺激减少85%，BFU-E和CFU-E来源的集落分别减少48%和78%。相反，反义处理的单核细胞经G-CSF刺激的粒细胞集落数量与对照组相比没有变化，尽管组成这些集落的细胞数量减少了大约90%。将c-myb反义寡聚体作用于my10+细胞，也得到了类似的结果。与未经处理的对照组相比，来自My10+细胞的CFU-GM和BFU-E集落数量没有变化，但组成这些集落的细胞数量与对照组相比分别减少了约75%和90%以上。C-myc正义和反义寡聚物对上述检测结果无明显影响。利用逆转录聚合酶链式反应，在0-8天的发育中的造血细胞中检测到c-myb基因的表达。在第14天，使用这种技术不再能检测到c-myb的表达。这些结果表明，c-myb是中晚期髓系和红系祖细胞增殖所必需的，但对谱系承诺和早期祖细胞扩增不那么重要。
To determine if MYB protein is preferentially required during specific stages of normal human hematopoiesis we incubated normal marrow mononuclear cells (MNC) with c-myb antisense oligodeoxynucleotides. Treated cells were cultured in semisolid medium under conditions designed to favor the growth of specific progenitor cell types. Compared with untreated controls, granulocyte-macrophage (GM) CFU-derived colonies decreased 77% when driven by recombinant human (rH) IL-3, and 85% when stimulated by rH GM colony-stimulating factor (CSF); erythroid burst-forming unit (BFU-E)- and CFU-E-derived colonies decreased 48 and 78%, respectively. In contrast, numbers of G-CSF-stimulated granulocyte colonies derived from antisense treated MNC were unchanged from controls, though the numbers of cells composing these colonies decreased approximately 90%. Similar results were obtained when MY10+ cells were exposed to c-myb antisense oligomers. When compared with untreated controls, numbers of CFU-GM and BFU-E colonies derived from MY10+ cells were unchanged, but the numbers of cells composing these colonies were reduced approximately 75 and greater than 90%, respectively, in comparison with controls. c-myc sense and antisense oligomers were without significant effect in these assays. Using the reverse transcription-polymerase chain reaction, c-myb mRNA was detected in developing hematopoietic cells on days 0-8. At day 14 c-myb expression was no longer detectable using this technique. These results suggest that c-myb is required for proliferation of intermediate-late myeloid and erythroid progenitors, but is less important for lineage commitment and early progenitor cell amplification.Images