6-Formylindolo(3,2-b)Carbazole (FICZ) Modulates the Signalsome Responsible for RA-Induced Differentiation of HL-60 Myeloblastic Leukemia Cells.
6-Formylindolo(3,2-b)Carbazole (FICZ) Modulates the Signalsome Responsible for RA-Induced Differentiation of HL-60 Myeloblastic Leukemia Cells.
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DOI：
10.1371/journal.pone.0135668
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yen A
中科院分区：
文献类型：
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作者：
Bunaciu RP;Jensen HA;MacDonald RJ;LaTocha DH;Varner JD;Yen A
6-Formylindolo(3,2-b)carbazole （FICZ）是色氨酸的光产物，是一种内源性芳烃受体（AhR）的高亲和力配体。此前有报道称，在患者源性HL-60髓母细胞白血病细胞中，视黄酸（RA）诱导的分化是由含有c-Cbl和AhR的信号体驱动的。通过表达膜分化标记CD38和CD11b、细胞周期阻滞和功能分化标记、诱导氧化代谢来评估FICZ增强ra诱导的分化。此外，FICZ增加了一些ra诱导的信号体成员的表达，如c-Cbl、Vav1、Slp76、PI3K和Src家族激酶Fgr和Lyn。为了研究ra诱导分化的分子信号，我们使用FRET和聚类分析表征了被认为驱动分化的关键分子的关联。在这里，我们报告，通过FRET检测，AhR在FICZ + ra诱导的分化中与c-Cbl相互作用，而AhR与Cbl-b组成相互作用。此外，基于流式细胞术评估分化标志物和western blot检测信号因子的相关性分析显示，Cbl-b、p-p38α和pT390-GSK3β与其他已知的ra诱导的信号成分或表型结果无关。我们注意到FICZ + RA引发的信号反应不是RA单独的典型信号反应，但可能代表了另一种分化驱动途径。在与细胞分化有关的信号分子簇中，FICZ与RA共同作用增强了RA诱导的动态变化的类型和强度。我们的数据表明FICZ与分化诱导治疗相关。其作用机制包括SFK和MAPK中心信号体的调节以及c- cl - ahr的关联。
6-Formylindolo(3,2-b)carbazole (FICZ) is a photoproduct of tryptophan and an endogenous high affinity ligand for aryl hydrocarbon receptor (AhR). It was previously reported that, in patient-derived HL-60 myeloblastic leukemia cells, retinoic acid (RA)-induced differentiation is driven by a signalsome containing c-Cbl and AhR. FICZ enhances RA-induced differentiation, assessed by expression of the membrane differentiation markers CD38 and CD11b, cell cycle arrest and the functional differentiation marker, inducible oxidative metabolism. Moreover, FICZ augments the expression of a number of the members of the RA-induced signalsome, such as c-Cbl, Vav1, Slp76, PI3K, and the Src family kinases Fgr and Lyn. Pursuing the molecular signaling responsible for RA-induced differentiation, we characterized, using FRET and clustering analysis, associations of key molecules thought to drive differentiation. Here we report that, assayed by FRET, AhR interacts with c-Cbl upon FICZ plus RA-induced differentiation, whereas AhR constitutively interacts with Cbl-b. Moreover, correlation analysis based on the flow cytometric assessment of differentiation markers and western blot detection of signaling factors reveal that Cbl-b, p-p38α and pT390-GSK3β, are not correlated with other known RA-induced signaling components or with a phenotypic outcome. We note that FICZ plus RA elicited signaling responses that were not typical of RA alone, but may represent alternative differentiation-driving pathways. In clusters of signaling molecules seminal to cell differentiation, FICZ co-administered with RA augments type and intensity of the dynamic changes induced by RA. Our data suggest relevance for FICZ in differentiation-induction therapy. The mechanism of action includes modulation of a SFK and MAPK centered signalsome and c-Cbl-AhR association.