[Different impacts of DMSO and RA on GPI-80 expression].

Shi yan sheng wu xue bao Pub Date : 2005-02-01
Jun Fen Fu, Yuji Takeda, Fujiro Sendo, Yoshihiko Araki
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Abstract

Our previous studies suggested that GPI-80 expression might be associated with myeloid differentiation. Dimethyl sulfoxide (DMSO) and all-trans-retinoic acid (RA) are two main inducers that can induce HL-60 cell to differentiate down the way of neutrophilic lineage. GPI-80 is a novel member of a human glycosylphosphatidylinositol (GPI)-anchored family that has been proposed as a potential regulator of beta2-integrin-dependent leukocyte adhesion, and is expressed mainly in human neutrophils. RT-PCR, flow cytometry and western-blot assays were used to detect the GPI-80 expression on HL-60 cells induced by DMSO or RA and to analyze the relationship between GPI-80 expression and CD11b or transferrin receptor (CD71) expression. The results showed GPI-80 was slightly expressed on the level of mRNA on RA-induced HL-60 cells by RT-PCR, while using flow cytometry and western-blot assays, We could not detect any GPI-80 expression on these cells, Furthermore, RA suppressed the induction of GPI-80 expression. However, DMSO could clearly derive the GPI-80 expression and this expression correlated well with the increasing dose and duration of DMSO stimulation. GPI-80 positive cells, which were included in CD11b-strongly positive and transferrin receptor-completely negative populations, were sufficiently differentiated cells suggesting that GPI-80 expression is tightly associated with the neutrophilic maturation and can be used as a mature neutrophilic marker. RA inhibits the GPI-80 expression on DMSO induced HL-60 cells suggests that different signal conduction has been activated by DMSO and RA when they induce the HL-60 cells to the neutrophilic differentiation.

[DMSO和RA对GPI-80表达的不同影响]。
我们之前的研究表明GPI-80的表达可能与骨髓分化有关。二甲基亚砜(DMSO)和全反式维甲酸(RA)是诱导HL-60细胞向中性粒细胞谱系分化的两种主要诱导剂。GPI-80是人糖基磷脂酰肌醇(GPI)锚定家族的新成员,已被认为是β -整合素依赖性白细胞粘附的潜在调节剂,主要在人中性粒细胞中表达。采用RT-PCR、流式细胞术和western-blot检测DMSO或RA诱导HL-60细胞GPI-80的表达,分析GPI-80表达与CD11b或转铁蛋白受体(CD71)表达的关系。RT-PCR结果显示,GPI-80在RA诱导的HL-60细胞mRNA水平上有少量表达,而流式细胞术和western-blot均未检测到GPI-80在HL-60细胞上的表达,RA抑制了GPI-80的诱导表达。而DMSO可以明显诱导GPI-80的表达,且GPI-80的表达与DMSO刺激剂量和持续时间的增加有良好的相关性。GPI-80阳性细胞包括在cd11b强阳性和转铁蛋白受体完全阴性的群体中,它们是充分分化的细胞,这表明GPI-80的表达与中性粒细胞成熟密切相关,可以作为成熟的中性粒细胞标志物。RA抑制DMSO诱导HL-60细胞GPI-80的表达,提示DMSO和RA在诱导HL-60细胞向嗜中性细胞分化时激活了不同的信号传导。
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