miR-454-3p 通过靶向 ZEB2 并调节 AKT/mTOR 通路促进 AML 细胞凋亡和自噬。

MiR-454-3p promotes apoptosis and autophagy of AML cells by targeting ZEB2 and regulating AKT/mTOR pathway.

机构信息

Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing, People's Republic of China.

Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, People's Republic of China.

出版信息

Hematology. 2023 Dec;28(1):2223874. doi: 10.1080/16078454.2023.2223874.

DOI:10.1080/16078454.2023.2223874

PMID:37313984

Abstract

BACKGROUND

miR-454-3p is considered to have a crucial role in cancer progression, but the potential involvement in acute myeloid leukemia (AML) remains unclear.

METHODS

Expression of miR-454-3p and ZEB2 mRNA and protein were quantified in AML cell lines. Cells were transfected with miR-454-3p inhibitor or mimic and cell growth was assessed by colony formation and CCK-8 assays and the cell cycle, apoptosis and autophagy were investigated by Western blotting, flow cytometry, immunofluorescence and 3-methyladenine (3-MA) treatment.

RESULTS

miR-454-3p expression was attenuated in AML cells. miR-454-3p overexpression attenuated cell growth and stimulated cell cycle arrest, apoptosis and autophagy. Dual-luciferase reporter assays and bioinformatics analysis showed that AML progression was inhibited when miR-454-3p regulated ZEB2, an effect confirmed by rescue assays. 3-MA reduced the autophagy-inducing effect of ZEB2 knockdown and indicated that autophagy induced apoptosis. miR-454-3p downregulated p-mTOR/p-AKT levels in AML cells.

CONCLUSION

The novel role of miR-454-3p as a tumor inhibitor in AML via regulation of the ZEB2/AKT/mTOR axis was demonstrated, indicating miR-454-3p as a potential new molecular target for AML.

摘要

背景

miR-454-3p 被认为在癌症进展中具有关键作用，但它在急性髓系白血病（AML）中的潜在作用尚不清楚。

方法

在 AML 细胞系中定量检测 miR-454-3p 和 ZEB2 mRNA 和蛋白的表达。用 miR-454-3p 抑制剂或模拟物转染细胞，通过集落形成和 CCK-8 测定评估细胞生长，通过 Western blot、流式细胞术、免疫荧光和 3-甲基腺嘌呤（3-MA）处理研究细胞周期、凋亡和自噬。

结果

miR-454-3p 在 AML 细胞中的表达减弱。miR-454-3p 的过表达减弱了细胞生长并刺激了细胞周期停滞、凋亡和自噬。双荧光素酶报告基因检测和生物信息学分析表明，当 miR-454-3p 调节 ZEB2 时，AML 进展受到抑制，通过挽救实验证实了这一作用。3-MA 减少了 ZEB2 敲低诱导的自噬，并表明自噬诱导了凋亡。miR-454-3p 下调了 AML 细胞中 p-mTOR/p-AKT 水平。