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环状 RNA circRNA_0000094 吸附 microRNA-223-3p 并上调 F-box 和 WD 重复域蛋白 7 来抑制 T 细胞急性淋巴细胞白血病进展。

Circular RNA circRNA_0000094 sponges microRNA-223-3p and up-regulate F-box and WD repeat domain containing 7 to restrain T cell acute lymphoblastic leukemia progression.

机构信息

Department of Pediatrics, Zhongnan Hospital of Wuhan University, No. 169 Donghu Road, Wuchang District, Wuhan, 430071, Hubei, China.

Department of Pediatrics, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, No. 136 Jingzhou Street, Xiangcheng District, Xiangyang City, 441021, Hubei Province, China.

出版信息

Hum Cell. 2021 May;34(3):977-989. doi: 10.1007/s13577-021-00504-4. Epub 2021 Mar 6.

DOI:10.1007/s13577-021-00504-4
PMID:33677796
Abstract

Circular RNAs (circRNAs) exert crucial regulatory effects in the pathogenesis of multiple tumors. This work aimed to probe into the role of circ_0000094 in T cell acute lymphoblastic leukemia (T-ALL). In this work, quantitative real-time polymerase chain reaction (qRT-PCR) was applied to quantify circ_0000094, miR-223-3p, and F-box and WD repeat domain containing 7 (FBW7) mRNA expressions in lymph node samples from T-ALL patients; Western blot was adopted to examine FBW7 protein expression in T-ALL cells; cell proliferation was detected by cell counting kit-8 (CCK-8) experiment; apoptosis was examined by flow cytometry; Transwell experiments were applied to assess T-ALL cell migration and invasion; the interactions among circ_0000094 and miR-223-3p, and miR-223-3p and FBW7 were validated by bioinformatics prediction, dual-luciferase reporter gene assay, and RNA immunoprecipitation experiment. We reported that, circ_0000094 expression was markedly reduced in T-ALL and circ_0000094 was predominantly located in the cytoplasm; gain-of-function and loss-of-function assays verified that circ_0000094 overexpression remarkably suppressed T-ALL cell proliferation, migration, and invasion, and enhanced apoptosis while knocking down circ_0000094 enhanced the malignant phenotypes of T-ALL cells; "rescue experiments" implied that miR-223-3p mimics partly reversed the inhibitory effects on the malignant phenotype of T-ALL cells due to the circ_0000094 up-regulation; circ_0000094 was proved to be a molecular sponge for miR-223-3p, and it could up-regulate the expression of FBW7 via repressing miR-223-3p expression. Taken together, it was concluded that circ_0000094 impedes T-ALL progression by modulating the miR-223-3p/FBW7 axis.

摘要

环状 RNA(circRNAs)在多种肿瘤的发病机制中发挥着关键的调节作用。本研究旨在探讨 circ_0000094 在 T 细胞急性淋巴细胞白血病(T-ALL)中的作用。本研究采用实时定量聚合酶链反应(qRT-PCR)检测 T-ALL 患者淋巴结样本中 circ_0000094、miR-223-3p 和 F-box 和 WD 重复域包含 7(FBW7)mRNA 的表达;采用 Western blot 检测 T-ALL 细胞中 FBW7 蛋白的表达;采用细胞计数试剂盒-8(CCK-8)实验检测细胞增殖;采用流式细胞术检测细胞凋亡;Transwell 实验评估 T-ALL 细胞迁移和侵袭;通过生物信息学预测、双荧光素酶报告基因检测和 RNA 免疫沉淀实验验证 circ_0000094 与 miR-223-3p 以及 miR-223-3p 与 FBW7 之间的相互作用。我们报道,circ_0000094 在 T-ALL 中表达显著降低,circ_0000094 主要位于细胞质中;功能获得和功能丧失实验验证了 circ_0000094 过表达显著抑制 T-ALL 细胞增殖、迁移和侵袭,并促进凋亡,而敲低 circ_0000094 则增强了 T-ALL 细胞的恶性表型;“挽救实验”表明,miR-223-3p 模拟物部分逆转了由于 circ_0000094 上调而对 T-ALL 细胞恶性表型的抑制作用;circ_0000094 被证明是 miR-223-3p 的分子海绵,通过抑制 miR-223-3p 的表达而上调 FBW7 的表达。总之,circ_0000094 通过调节 miR-223-3p/FBW7 轴阻碍 T-ALL 的进展。

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