The CGRP Receptor Antagonist MK0974 Induces EVI1high AML Cell Apoptosis by Disrupting ERK Signaling
The CGRP Receptor Antagonist MK0974 Induces EVI1high AML Cell Apoptosis by Disrupting ERK Signaling
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DOI：
10.21873/anticanres.15979
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发表时间：
2022-10
影响因子：
2
通讯作者：
Akira Suekane;T. Ichikawa;Yusuke Saito;S. Nakahata;K. Morishita
中科院分区：
文献类型：
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作者：
Akira Suekane;T. Ichikawa;Yusuke Saito;S. Nakahata;K. Morishita
摘要背景/目的：高表达致癌转录因子亲嗜性病毒整合位点-1（EVI 1 high AML）的急性髓细胞白血病（AML）难治，急需开发EVI 1 high AML的治疗方法。我们先前的研究表明，降钙素受体样受体（CRLR）/受体活性修饰蛋白1（RAMP 1）在EVI 1高AML中高度表达，并参与降钙素基因相关肽（CGRP）诱导的应激性造血。本研究检测了MK 0974（一种CGRP拮抗剂）是否在CRLR/RAMP 1high AML细胞系中作为治疗剂。材料与方法：采用体外实验系统，观察MK 0974对EVI 1high AML细胞系的作用。逆转录聚合酶链反应（RT-PCR）检测CRLR和RAMP 1 -3在EVI 1high和EVI 1 low AML细胞系中的表达。接下来，将MK 0974加入AML细胞系中，并使用流式细胞术（FCM）进行细胞增殖、细胞周期和凋亡测定。使用蛋白质印迹分析评价蛋白质。我们还产生了CRLR敲低的AML细胞系，并评估了MK 0974的作用是否降低。结果：MK 0974可诱导EVI 1high AML细胞凋亡。在EVI 1high AML细胞系中，添加MK 0974减弱了ERK和p38的磷酸化。CRLR敲除也减弱了这些作用。结论：CGRP受体拮抗剂MK 0974可抑制CRLR/RAMP 1复合物并诱导细胞凋亡，有望成为CRLR/RAMP 1high AML的治疗药物。
Abstract Background/Aim: Acute myeloid leukemia (AML) with high expression of the oncogenic transcription factor ecotropic viral integration site-1 (EVI1) (EVI1high AML) is refractory, and there is an urgent need to develop treatment for EVI1high AML. We previously showed that calcitonin receptor-like receptor (CRLR)/receptor activity modifying protein 1 (RAMP1) is highly expressed in EVI1high AML and participates in calcitonin gene-related peptide (CGRP)-induced stress hematopoiesis. This study examined whether MK0974 (a CGRP antagonist) acts as a therapeutic agent in CRLR/RAMP1high AML cell lines. Materials and Methods: An in vitro experimental system was used to determine the effect of MK0974 on EVI1high AML cell lines. The expression of CRLR and RAMP1-3 in EVI1high and EVI1low AML lines was evaluated by reverse-transcription polymerase chain reaction (RT–PCR). Next, MK0974 was added to the AML cell lines, and cell proliferation, cell cycle and apoptosis assays were carried out using flow cytometry (FCM). Proteins were evaluated using western blot analysis. We also generated AML cell lines with CRLR knockdown and evaluated whether the effect of MK0974 was reduced. Results: Apoptosis was induced by adding MK0974 to the EVI1high AML cell line. In the EVI1high AML cell line, the addition of MK0974 attenuated the phosphorylation of ERK and p38. These effects were also attenuated by CRLR knockdown. Conclusion: MK0974, a CGRP receptor antagonist, inhibits the CRLR/RAMP1 complex and induces apoptosis, making it a potential therapeutic agent for CRLR/RAMP1high AML.