miR-29b-3p 通过靶向 HuR 抑制 NF-κB 和 JAK/STAT 信号通路来抑制 AML 细胞的恶性生物学行为。
miR-29b-3p suppresses the malignant biological behaviors of AML cells via inhibiting NF-κB and JAK/STAT signaling pathways by targeting HuR.
机构信息
Department of Laboratory Medicine, The School of Medical Technology and Engineering, Fujian Medical University, Fuzhou, 350004, Fujian, China.
Department of Dermatology, The Third People's Hospital of Hangzhou, Hangzhou, 310009, Zhejiang, China.
出版信息
BMC Cancer. 2022 Aug 20;22(1):909. doi: 10.1186/s12885-022-09996-1.
BACKGROUND
HuR/ELAVL1 (embryonic lethal abnormal vision 1) was a downstream target of miR-29b in some cancer cells. HuR protein exerts important prognostic effects of involving in the pathogenesis and development of acute myeloid leukemia (AML). This study aims to investigate the role of miR-29b-3p in biological behaviors of AML cells by targeting HuR and the involvement of the NF-κB and JAK/STAT signaling pathways.
METHODS
The expressions of HuR and miR-29b-3p in AML cells were determined using RT-qPCR and Western blot, and the association between them was analyzed using the Spearman method. Next, the target relationship between HuR and miR-29b-3p was predicted by biological information databases and verified by the dual-luciferase reporter gene assay. MTS, methyl cellulose, flow cytometry and transwell assay were employed to detect the cell proliferation, clone formation, cell cycle and apoptosis, invasion and migration respectively, the effect of miR-29b-3p targeted HuR on the biological behaviors of AML cells was explored after over- /down-expression of miR-29b-3p and rescue experiment. Then, immunofluorescence assay and western blot were employed to detect location expression and phosphorylation levels of NF-κB and JAK/STAT signaling pathways related molecules respectively.
RESULTS
HuR was negatively correlated with miR-29b-3p, and was the downstream target of miR-29b-3p in AML cells. When miR-29b-3p was overexpressed in AML cells, HuR was down-regulated, accompanied by cell viability decreased, cell cycle arrest, apoptosis increased, invasion and migration weakened. Moreover, the opposite result appeared after miR-29b-3p was down-regulated. The rescue experiment showed that miR-29b-3p inhibitor could reverse the biological effect of HuR down-regulation in AML cells. Molecular pathway results showed that miR-29b-3p could inhibit p65 expression in nucleus and phosphorylation levels of p65, IκBα, STAT1, STAT3 and STAT5.
CONCLUSION
miR-29b-3p can inhibit malignant biological behaviors of AML cells via the inactivation of the NF-κB and JAK/STAT signaling pathways by targeting HuR. miR-29b-3p and its target HuR can be used as a new potential molecular for AML treatment.
背景
在某些癌细胞中，HuR/ELAVL1（胚胎致死异常视觉 1）是 miR-29b 的下游靶标。HuR 蛋白在急性髓细胞白血病（AML）的发病机制和发展中具有重要的预后作用。本研究旨在通过靶向 HuR 探讨 miR-29b-3p 在 AML 细胞生物学行为中的作用，以及 NF-κB 和 JAK/STAT 信号通路的参与情况。
方法
采用 RT-qPCR 和 Western blot 检测 AML 细胞中 HuR 和 miR-29b-3p 的表达情况，采用 Spearman 法分析两者之间的相关性。然后，通过生物信息数据库预测 HuR 与 miR-29b-3p 的靶标关系，并通过双荧光素酶报告基因检测进行验证。采用 MTS、甲基纤维素、流式细胞术和 Transwell 实验分别检测细胞增殖、克隆形成、细胞周期和凋亡、侵袭和迁移情况，探讨过表达/下调 miR-29b-3p 及挽救实验后 miR-29b-3p 靶向 HuR 对 AML 细胞生物学行为的影响。然后，采用免疫荧光和 Western blot 检测 NF-κB 和 JAK/STAT 信号通路相关分子的定位表达和磷酸化水平。
结果
HuR 与 miR-29b-3p 呈负相关，且为 AML 细胞中 miR-29b-3p 的下游靶标。当 AML 细胞中 miR-29b-3p 过表达时，HuR 下调，细胞活力下降，细胞周期阻滞，凋亡增加，侵袭和迁移减弱。反之，下调 miR-29b-3p 时则出现相反的结果。挽救实验表明，miR-29b-3p 抑制剂可逆转 HuR 下调对 AML 细胞生物学作用。分子通路结果表明，miR-29b-3p 可抑制核内 p65 表达及其磷酸化水平，以及 IκBα、STAT1、STAT3 和 STAT5。
结论
miR-29b-3p 可通过靶向 HuR 抑制 NF-κB 和 JAK/STAT 信号通路的活化，抑制 AML 细胞的恶性生物学行为。miR-29b-3p 及其靶标 HuR 可作为 AML 治疗的新潜在分子。
Zotero 插件