hsa_circ_0121582 inhibits leukemia growth by dampening Wnt/β-catenin signaling
hsa_circ_0121582 inhibits leukemia growth by dampening Wnt/β-catenin signaling
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DOI：
10.1007/s12094-020-02377-9
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发表时间：
2020-05-29
影响因子：
3.4
通讯作者：
Zhou, M.
中科院分区：
文献类型：
--
作者：
Chen, J-J.;Lei, P.;Zhou, M.
目的化疗后的AML患者预后较差，尤其是对化疗药物不敏感和耐药的患者。为了阐明AML的潜在发病机制，为临床治疗提供新的治疗靶点，我们探讨了circRNA在白血病中的作用。方法对白血病患者和健康捐献者进行高通量circRNA测序分析。 RT-qPCR 和蛋白质印迹分析用于确定 GSK3 beta 的表达。 RNA Pull-down 测定用于检测 hsa_circ_0121582 下拉的 miRNA。采用RNA免疫沉淀法评估TET1与hsa_circ_0121582之间的结合能力。结果发现一种新的、高度稳定的circRNA，其源自GSK3 beta外显子1至外显子7的反向剪接，并且hsa_circ_0121582在白血病细胞中下调。在功能获得实验中，上调的hsa_circ_0121582在体外和体内抑制白血病细胞的增殖。在细胞质中，hsa_circ_0121582可以充当miR-224的海绵，减弱miR-224对GSK3β的抑制作用，从而上调GSK3β的表达水平。此外，hsa_circ_0121582可以与细胞核中的GSK3 beta启动子结合，并招募DNA去甲基化酶TET1以确保GSK3 beta的转录。上调的GSK3β抑制Wnt/β-catenin信号通路，减少β-catenin在细胞核内的聚集，从而抑制白血病细胞的增殖。结论本研究发现hsa_circ_0121582参与抑制肿瘤增殖，并恢复hsa_circ_0121582 可能是白血病患者的有效治疗策略。
Purpose The prognosis of AML patients with chemotherapy is poor, especially those who are insensitive to and resistant to chemotherapy drugs. To clarify the underlying pathogenesis of AML and provide new therapeutic targets for clinical treatment, we explore the role of circRNA in leukemia.Methods High-throughput circRNA sequencing analysis was performed in patients with leukemia and healthy donors. RT-qPCR and western blot analysis were used to determine expression of GSK3 beta. RNA pull-down assay was used to detect miRNAs pulled down by hsa_circ_0121582. RNA immunoprecipitation assay was performed to evaluate the binding capacity between TET1 and hsa_circ_0121582.Results A new and highly stable circRNA was found, which was derived from the reverse splicing of GSK3 beta exon 1 to exon 7, and hsa_circ_0121582 was down-regulated in leukemia cells. In gain-of-function experiments, the up-regulated hsa_circ_0121582 inhibited the proliferation of leukemia cells in vitro and in vivo. In the cytoplasm, hsa_circ_0121582 could act as a sponge for miR-224, attenuate the inhibiting effect of miR-224 on GSK3 beta, and thus up-regulate the expression level of GSK3 beta. In addition, hsa_circ_0121582 could bind to GSK3 beta promoter in the nucleus, and recruit DNA demethylase TET1 to ensuring the transcription of GSK3 beta. The upregulated GSK3 beta inhibited the Wnt/beta-catenin signaling pathway, and reduced the aggregation of beta-catenin in the nucleus, thus inhibited the proliferation of leukemia cells.Conclusions This study found that hsa_circ_0121582 was involved in the inhibition of tumor proliferation, and the restoration of hsa_circ_0121582 could be an effective treatment strategy for patients with leukemia.