The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors
The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors
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DOI：
10.1182/blood.v99.2.507
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发表时间：
2002-01-15
期刊：
影响因子：
20.3
通讯作者：
Torok-Storb, B
中科院分区：
文献类型：
--
作者：
Scharenberg, CW;Harkey, MA;Torok-Storb, B
造血干细胞的一个有前途的和越来越多的开发性质是它们流出荧光染料Hoechst 33342的能力。Hoechst阴性细胞通过荧光激活细胞分选分离为所谓的骨髓侧“群”（SP）。据报道，来自骨髓以及其他组织的SIR含有具有相当大可塑性的未成熟干细胞。一些细胞系还外排Hoechst并产生SP谱。逆转录-聚合酶链反应（RT-PCR）和外排抑制研究与肺癌细胞系，A549，牵连的ABCG 2转运作为Hoechst外排泵。此外，显示ABCG 2的瞬时表达在人胚肾（HEK 293）细胞中产生稳健的SP表型。结果允许ABCG 2是有效的Hoechst外排泵的结论。采用半定量RT-PCR方法研究ABCG 2在造血过程中的表达规律。它在假定的造血干细胞（分离为SP、34（+）/38（-）或34（+）/KDR+群体）中以相对高的水平表达，并且在定向祖细胞中急剧下降（34（+）/38（+）、34（+）/33（+）或34（+）/10（+））。在大多数成熟人群中表达仍然较低，但在自然杀伤细胞和成红细胞中再次升高。比较骨髓SP细胞中3种主要多药耐药外排泵MDR 1、MRP 1和ABCG 2的信使RNA（mRNA）水平，发现ABCG 2是这些细胞中的主要形式。这些数据表明，ABCG 2显着有助于造血干细胞中SP表型的产生。此外，ABCG 2在谱系定型阶段的急剧下调表明该基因可能在多能干细胞的独特生理学中发挥重要作用。（血。2002; 99：507-512）（C）2002年由美国血液学会。
A promising and increasingly exploited property of hematopoietic stem cells is their ability to efflux the fluorescent dye Hoechst 33342. The Hoechst-negative cells are isolated by fluorescence-activated cell sorting as a so-called side "population" (SP) of bone marrow. This SIR from bone marrow, as well as other tissues, is reported to contain immature stem cells with considerable plasticity. Some cell lines also efflux Hoechst and generate SP profiles. Reverse transcription-polymerase chain reaction (RT-PCR) and efflux inhibition studies with the lung carcinoma cell line, A549, implicated the ABCG2 transporter as a Hoechst efflux pump. Furthermore, it is shown that transient expression of ABCG2 generates a robust SP phenotype in human embryonic kidney (HEK293) cells. The results allow the conclusion that ABCG2 is a potent Hoechst efflux pump. Semiquantitative RT-PCR was used to characterize the developmental pattern of expression of ABCG2 in hematopoiesis. It is expressed at relatively high levels in putative hematopoietic stem cells (isolated as SP, 34(+)/38(-) or 34(+)/KDR+ populations) and drops sharply in committed progenitors (34(+)/38(+), 34(+)/33(+), or 34(+)/10(+)). Expression remains low in most maturing populations, but rises again in natural killer cells and erythroblasts. Comparison of messenger RNA (mRNA) levels for the 3 major multidrug-resistant efflux pumps, MDR1, MRP1, and ABCG2, in bone marrow SP cells reveals that ABCG2 is the predominant form in these cells. These data suggest that ABCG2 contributes significantly to the generation of the SP phenotype in hematopoietic stem cells. Furthermore, the sharp down-regulation of ABCG2 at the stage of lineage commitment suggests that this gene may play an important role in the unique physiology of the pluripotent stem cell. (Blood. 2002; 99:507-512) (C) 2002 by The American Society of Hematology.