环状非编码 RNA NFIX 敲低通过 miR-876-3p/TRIM31 轴抑制 AML 致瘤性。

CircNFIX knockdown inhibited AML tumorigenicity by the miR-876-3p/TRIM31 axis.

机构信息

Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

Department of Blood Transfusion, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

出版信息

Hematology. 2022 Dec;27(1):1046-1055. doi: 10.1080/16078454.2022.2115699.

DOI:10.1080/16078454.2022.2115699

PMID:36094501

Abstract

BACKGROUND

Acute myeloid leukemia (AML) is one of the most common malignant myeloid diseases in adults with a dismal prognosis. We aimed to explore the effects of circNFIX on the proliferation and apoptosis of AML cells.

METHODS

The expressions of circNFIX, miR-876-3p and tripartite motif (TRIM) 31 in the bone marrow specimens of AML patients and AML cell lines were detected by qRT-PCR or western blot. Cell proliferation was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-ethynyl-29-deoxyuridine (EdU) assays. Cell cycle and apoptosis were analyzed by flow cytometry. Western blot was used to detect protein expression. The relationship between miR-876-3p and circNFIX or TRIM31 was identified by dual-luciferase reporter assay or RNA pull-down assay.

RESULTS

The expression level of circNFIX was significantly increased in the bone marrow samples of AML patients and AML cells when compared with normal controls. CircNFIX silencing inhibited AML cell proliferation and promoted apoptosis. Inhibition of miR-876-3p reversed the effect of circNFIX knockdown on AML cell progression. In addition, circNFIX indirectly regulated TRIM31 through miR-876-3p. Further, TRIM31 overexpression counteracted the effect of circNFIX silencing on AML cell proliferation and apoptosis.

CONCLUSION

CircNFIX knockdown could suppress the proliferation and induce the apoptosis of AML cells by targeting the miR-876-3p/TRIM31 axis.

摘要

背景

急性髓系白血病（AML）是成人中最常见的恶性髓系疾病之一，预后不良。我们旨在探讨 circNFIX 对 AML 细胞增殖和凋亡的影响。

方法

通过 qRT-PCR 或 Western blot 检测 AML 患者和 AML 细胞系骨髓标本中 circNFIX、miR-876-3p 和三肽基（TRIM）31 的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）和 5-乙炔基-29-脱氧尿苷（EdU）检测细胞增殖。通过流式细胞术分析细胞周期和细胞凋亡。Western blot 用于检测蛋白表达。通过双荧光素酶报告基因检测或 RNA 下拉实验确定 miR-876-3p 与 circNFIX 或 TRIM31 的关系。

结果

与正常对照组相比，AML 患者和 AML 细胞的骨髓样本中 circNFIX 的表达水平显著升高。circNFIX 沉默抑制 AML 细胞增殖并促进凋亡。抑制 miR-876-3p 逆转了 circNFIX 敲低对 AML 细胞进展的影响。此外，circNFIX 通过 miR-876-3p 间接调节 TRIM31。进一步，TRIM31 过表达抵消了 circNFIX 沉默对 AML 细胞增殖和凋亡的影响。