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环状 RNA-UBE2D2 通过调控 microRNA-376a-3p/真核翻译起始因子 4γ2 轴促进非小细胞肺癌细胞的增殖和转移。

Circular RNA-UBE2D2 accelerates the proliferation and metastasis of non-small cell lung cancer cells via modulating microRNA-376a-3p/Eukaryotic Translation Initiation Factor 4γ2 axis.

机构信息

Department of Cardiothoracic Surgery, Changzhou Second People's Hospital, Changzhou City, Jiangsu Province, China.

出版信息

Bioengineered. 2022 Mar;13(3):5942-5953. doi: 10.1080/21655979.2022.2027068.

Abstract

Non-small cell lung cancer (NSCLC) ranks first in the morbidity and mortality of malignant tumors in China. As reported, circular RNAs (circRNAs) are emerged in the progress of NSCLC. The study was to figure out the potential mechanism of circ-UBE2D2 in the progression of NSCLC. First, plasmid vectors intervening circ-UBE2D2, microRNA (miR)-376a-3p or Eukaryotic Translation Initiation Factor 4γ2 (EIF4G2) expression were transfected into NSCLC cells, and the expression of circ-UBE2D2, miR-376a-3p and EIF4G2 was detected by reverse transcription quantitative polymerase chain reaction or Western blot. Then, cell proliferation was detected by Cell counting kit-8 assay and plate cloning. Cell apoptosis was tested by flow cytometry. Plate scratches and Transwell were used to detect cell migration and invasion. Finally, the binding sites of circRNA UBE2D2, EIF4G2 and miR-376a-3p were verified by bioinformatics website starBase analysis and dual luciferase reporter gene assay. The results manifested the up-regulation of circ-UBE2D2 expression in NSCLC tissues and cells. Circ-UBE2D2 promoted the proliferation, migration and invasion, but repressed apoptosis of NSCLC cells. Interestingly, circ-UBE2D2 directly targeted miR-376a-3p and up-regulated miR-376a-3p restrained proliferation, migration and invasion, but accelerated apoptosis of NSCLC cells. More importantly, EIF4G2 was the target of miR-376a-3p, and overexpression of EIF4G2 reversed the effects of circ-UBE2D2 downregulation on proliferation, migration, invasion and apoptosis of NSCLC cells. These results suggest that circ-UBE2D2 promotes the proliferation, migration and invasion but restrains apoptosis of lung cancer cells by regulating miR-376a-3p/EIF4G2 axis.

摘要

非小细胞肺癌(NSCLC)在中国恶性肿瘤的发病率和死亡率中均位居首位。据报道,环状 RNA(circRNAs)在 NSCLC 的发生发展中逐渐显现。本研究旨在探讨 circ-UBE2D2 在 NSCLC 进展中的潜在机制。首先,将干预 circ-UBE2D2、microRNA(miR)-376a-3p 或真核翻译起始因子 4γ2(EIF4G2)表达的质粒载体转染入 NSCLC 细胞中,通过逆转录定量聚合酶链反应或 Western blot 检测 circ-UBE2D2、miR-376a-3p 和 EIF4G2 的表达。然后,通过细胞计数试剂盒-8 检测细胞增殖,平板克隆检测细胞增殖,流式细胞术检测细胞凋亡,平板划痕和 Transwell 检测细胞迁移和侵袭。最后,通过生物信息学网站 starBase 分析和双荧光素酶报告基因检测验证 circRNA UBE2D2、EIF4G2 和 miR-376a-3p 的结合位点。结果表明,circ-UBE2D2 在 NSCLC 组织和细胞中呈上调表达。Circ-UBE2D2 促进 NSCLC 细胞的增殖、迁移和侵袭,但抑制其凋亡。有趣的是,circ-UBE2D2 可直接靶向 miR-376a-3p,上调 miR-376a-3p 可抑制 NSCLC 细胞的增殖、迁移和侵袭,但促进其凋亡。更重要的是,EIF4G2 是 miR-376a-3p 的靶点,过表达 EIF4G2 可逆转 circ-UBE2D2 下调对 NSCLC 细胞增殖、迁移、侵袭和凋亡的作用。这些结果提示,circ-UBE2D2 通过调节 miR-376a-3p/EIF4G2 轴促进肺癌细胞的增殖、迁移和侵袭,但抑制其凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f282/8974110/ad67d2e86e0f/KBIE_A_2027068_F0001_OC.jpg

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