苔藓虫素1诱导急性淋巴细胞白血病细胞泛素cooh末端水解酶。

Enzyme & protein Pub Date : 1996-01-01 DOI:10.1159/000468636
R M Mohammad, A Maki, G R Pettit, A M al-Katib
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引用次数: 10

摘要

以前有报道称苔藓虫素1 (Bryo1)诱导人类急性淋巴细胞白血病(ALL)细胞系Reh分化为单核细胞b细胞阶段。在这项研究中,我们证明了一种新的蛋白质,泛素cooh末端水解酶(UCH-L1),与这种分化有关。用200 nmol/l Bryo1处理Reh细胞72 h,观察细胞形态、表面免疫表型、酸性磷酸酶和末端脱氧核苷酸转移酶的变化。通过二维聚丙烯酰胺凝胶电泳(2D PAGE)研究亲本细胞和分化细胞的蛋白质图谱。bryo1处理的细胞表达了单核细胞b细胞阶段的形态学、表型和酶学特征。在分化细胞中检测到UCH-L1酶(MW-pl 34-5.3),而在亲本细胞中未检测到。使用UCH-L1多克隆抗体对bryo1处理的细胞进行2 D PAGE免疫印迹,进一步证实UCH-L1存在于bryo1处理的细胞中。研究了泛素在亲本和bryo1处理细胞中的表达,并与TPA处理细胞进行了比较。流式细胞术检测,TPA和Bryo1均能提高泛素表达水平。作为UCH-L1的抑制剂,硼氢化钠抑制bryo1诱导的Reh细胞分化作用。迄今为止,Bryo1发挥其b细胞分化作用的机制尚不完全清楚。本研究表明UCH-L1的表达可能在bryo1诱导的b - all前分化中起重要作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Bryostatin 1 induces ubiquitin COOH-terminal hydrolase in acute lymphoblastic leukemia cells.

It has been previously reported that Bryostatin 1 (Bryo1) induces differentiation of the human acute lymphoblastic leukemia (ALL) cell line, Reh, to a monocytoid B-cell stage. In this study we demonstrate that a novel protein, ubiquitin COOH-terminal hydrolase (UCH-L1), is associated with this differentiation. Reh cells were treated with 200 nmol/l of Bryo1 for 72 h and analyzed for changes in morphology, surface immunophenotype, acid phosphatase and terminal deoxynucleotidyl transferase. Protein patterns of the parent and differentiated cells, by two-dimensional polyacrylamide gel electrophoresis (2D PAGE), were studied. Bryo1-treated cells expressed morphologic, phenotypic and enzymatic features of the monocytoid B-cell stage. The UCH-L1 enzyme (MW-pl 34-5.3) was detected by 2 D PAGE in the differentiated, but not in parent cells. The presence of UCH-L1 in the Bryo1-treated cells was further confirmed by immunoblotting of 2 D PAGE using UCH-L1 polyclonal antibody. Ubiquitin expression was studied in parent and Bryo1-treated cells and was compared with 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated cells. Both agents, TPA and Bryo1, increased the level of ubiquitin expression as detected by flow cytometry. Sodium borohydride, an inhibitor of UCH-L1, inhibited the Bryo1-induced differentiating effect on Reh cells. To date, the mechanism by which Bryo1, exerts its B-cell differentiating effect is not fully understood. This study shows that UCH-L1 expression may play a major role in Bryo1-induced differentiation in pre-B-ALL.

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