Abstract：
BACKGROUND: Despite recent advances in therapeutic regimens, the prognosis of acute myeloid leukemia (AML) remains poor. Following our previous finding that interleukin-33 (IL-33) promotes cell survival along with activated NF-κB in AML, we further investigated the role of NF-κB during leukemia development.
METHODS: Flow cytometry was performed to value the apoptosis and proliferation. qRT-PCR and western blot were performed to detect the expression of IL-6, active caspase 3, BIRC2, Bcl-2, and Bax, as well as activated NF-κB p65 and AKT. Finally, xenograft mouse models and AML patient samples were used to verify the findings observed in AML cell lines.
RESULTS: IL-33-mediated NF-κB activation in AML cell lines contributes to a reduction in apoptosis, an increase in proliferation rate as well as a decrease in drug sensitivity, which were reversed by NF-κB inhibitor, Bay-117085. Moreover, IL-33 decreased the expression of active caspase-3 while increasing the levels of BIRC2, Bcl-2, and Bax, and these effects were blocked by Bay-117085. Additionally, NF-κB activation induced by IL-33 increases the production of IL-6 and autocrine activation of AKT. Co-culture of bone marrow stroma with AML cells resulted in increased IL-33 expression by leukemia cells, along with decreased apoptosis level and reduced drug sensitivity. Finally, we confirmed the in vivo pro-tumor effect mediated by IL-33/ NF-κB axis using a xenograft model of AML.
CONCLUSIONS: Our data indicate that IL-33/IL1RL1-dependent signaling contributes to AML cell activation of NF-κB, which in turn causes autocrine IL-6-induced activation of pAKT, supporting IL-33/NF-κB/pAKT as a potential target for AML therapy.
METHODS: Flow cytometry was performed to value the apoptosis and proliferation. qRT-PCR and western blot were performed to detect the expression of IL-6, active caspase 3, BIRC2, Bcl-2, and Bax, as well as activated NF-κB p65 and AKT. Finally, xenograft mouse models and AML patient samples were used to verify the findings observed in AML cell lines.
RESULTS: IL-33-mediated NF-κB activation in AML cell lines contributes to a reduction in apoptosis, an increase in proliferation rate as well as a decrease in drug sensitivity, which were reversed by NF-κB inhibitor, Bay-117085. Moreover, IL-33 decreased the expression of active caspase-3 while increasing the levels of BIRC2, Bcl-2, and Bax, and these effects were blocked by Bay-117085. Additionally, NF-κB activation induced by IL-33 increases the production of IL-6 and autocrine activation of AKT. Co-culture of bone marrow stroma with AML cells resulted in increased IL-33 expression by leukemia cells, along with decreased apoptosis level and reduced drug sensitivity. Finally, we confirmed the in vivo pro-tumor effect mediated by IL-33/ NF-κB axis using a xenograft model of AML.
CONCLUSIONS: Our data indicate that IL-33/IL1RL1-dependent signaling contributes to AML cell activation of NF-κB, which in turn causes autocrine IL-6-induced activation of pAKT, supporting IL-33/NF-κB/pAKT as a potential target for AML therapy.
摘要：
背景：尽管治疗方案有了最新进展，急性髓系白血病(AML)的预后仍然较差.根据我们先前的发现，白细胞介素-33（IL-33）促进AML中细胞存活以及活化的NF-κB，我们进一步研究了NF-κB在白血病发展过程中的作用。
方法：流式细胞术评估细胞凋亡和增殖。qRT-PCR和westernblot检测IL-6、活性caspase3、BIRC2、Bcl-2和Bax的表达,以及活化的NF-κBp65和AKT。最后,异种移植小鼠模型和AML患者样本用于验证在AML细胞系中观察到的发现.
结果：IL-33介导的NF-κB在AML细胞系中的激活有助于减少细胞凋亡，增殖率的增加以及药物敏感性的降低，被NF-κB抑制剂逆转，Bay-117085.此外，IL-33降低了活性caspase-3的表达，同时增加了BIRC2，Bcl-2和Bax的水平，这些影响被Bay-117085阻断。此外,IL-33诱导的NF-κB活化增加了IL-6的产生和AKT的自分泌活化。骨髓基质与AML细胞共培养导致白血病细胞IL-33表达增加，细胞凋亡水平降低，药物敏感性降低。最后,我们使用AML异种移植模型证实了IL-33/NF-κB轴介导的体内促肿瘤作用.
结论：我们的数据表明IL-33/IL1RL1依赖性信号传导有助于AML细胞NF-κB的激活，这反过来又导致自分泌IL-6诱导的pAKT激活，支持IL-33/NF-κB/pAKT作为AML治疗的潜在靶点。
方法：流式细胞术评估细胞凋亡和增殖。qRT-PCR和westernblot检测IL-6、活性caspase3、BIRC2、Bcl-2和Bax的表达,以及活化的NF-κBp65和AKT。最后,异种移植小鼠模型和AML患者样本用于验证在AML细胞系中观察到的发现.
结果：IL-33介导的NF-κB在AML细胞系中的激活有助于减少细胞凋亡，增殖率的增加以及药物敏感性的降低，被NF-κB抑制剂逆转，Bay-117085.此外，IL-33降低了活性caspase-3的表达，同时增加了BIRC2，Bcl-2和Bax的水平，这些影响被Bay-117085阻断。此外,IL-33诱导的NF-κB活化增加了IL-6的产生和AKT的自分泌活化。骨髓基质与AML细胞共培养导致白血病细胞IL-33表达增加，细胞凋亡水平降低，药物敏感性降低。最后,我们使用AML异种移植模型证实了IL-33/NF-κB轴介导的体内促肿瘤作用.
结论：我们的数据表明IL-33/IL1RL1依赖性信号传导有助于AML细胞NF-κB的激活，这反过来又导致自分泌IL-6诱导的pAKT激活，支持IL-33/NF-κB/pAKT作为AML治疗的潜在靶点。
