环状MYC通过miR-516a-5p/AKT3轴调节急性髓系白血病中的线粒体呼吸和细胞活力。
CircMYC regulates the mitochondrial respiration and cell viability miR-516a-5p/AKT3 axis in acute myeloid leukemia.
作者信息
Zou Xueqin, Jiang Min
机构信息
Health Management Center, Shenzhen University General Hospital Shenzhen 518055, Guangdong Province, China.
Department of Pediatrics, Shenzhen University General Hospital Shenzhen 518055, Guangdong Province, China.
出版信息
Am J Transl Res. 2021 Sep 15;13(9):10112-10126. eCollection 2021.
OBJECTIVE
Acute myeloid leukemia (AML) is a hematological malignancy with an aberrant proliferation of immature myeloid cells. This study aimed at exploring the regulatory function of circMYC in AML progression.
METHODS
Expression levels of CircMYC, miR-516a-5p, AKT3 and apoptosis-related proteins were determined by RT-qPCR and western blot. Cell viability and proliferation were examined by CCK8 assay and EdU assay. Annexin V/PI staining was used to assess cell apoptosis. Mitochondrial respiration function was confirmed by oxygen consumption rate (OCR). The relationships among circMYC, miR-516a-5p and AKT3 were detected by dual-luciferase reporter (DLR) assay, RNA-pull down assay and RNA immunoprecipitation (RIP) assay, respectively.
RESULTS
CircMYC was positively correlated with poor prognosis in AML patients (all P<0.05). Knockdown of circMYC decreased cell viability and OCR but increased cell apoptosis rates (all P<0.05), and miR-516a-5p overexpression displayed the similar trend. Mechanistically, the oncogenic effects of circMYC were achieved by sponging miR-516a-5p and increasing AKT3.
CONCLUSION
Decreased expression of circMYC could suppress AML progression by regulating miR-516a-5p/AKT3, suggesting a new therapeutic target in AML treatment.
目的
急性髓系白血病（AML）是一种血液系统恶性肿瘤，其特征为未成熟髓系细胞异常增殖。本研究旨在探讨环状MYC（circMYC）在AML进展中的调控作用。
方法
采用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测circMYC、微小RNA-516a-5p（miR-516a-5p）、蛋白激酶B3（AKT3）的表达水平及凋亡相关蛋白的表达。采用细胞计数试剂盒8（CCK8）法和5-乙炔基-2'-脱氧尿苷（EdU）法检测细胞活力和增殖情况。采用膜联蛋白V/碘化丙啶（Annexin V/PI）染色法评估细胞凋亡。通过耗氧率（OCR）检测线粒体呼吸功能。分别采用双荧光素酶报告基因（DLR）实验、RNA下拉实验和RNA免疫沉淀（RIP）实验检测circMYC、miR-516a-5p和AKT3之间的关系。
结果
circMYC与AML患者的不良预后呈正相关（均P<0.05）。敲低circMYC可降低细胞活力和OCR，但增加细胞凋亡率（均P<0.05），miR-516a-5p过表达也呈现类似趋势。机制上，circMYC的致癌作用是通过吸附miR-516a-5p并上调AKT3实现的。
结论
circMYC表达降低可通过调节miR-516a-5p/AKT3抑制AML进展，提示其可作为AML治疗的新靶点。
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