Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, 81, Meishan Road, Shushan District, Hefei City, 230000, Anhui Province, China.
Anhui Medical University School of Medicine, 81, Meishan Road, Shushan District, Hefei City, 230000, Anhui Province, China.
Mol Cell Biochem. 2022 Sep;477(9):2161-2171. doi: 10.1007/s11010-020-04026-9. Epub 2021 Jan 8.
Circular RNAs (circRNAs) have emerged as a multifunctional class of RNAs, while there is limited knowledge on their functions in the development of cancers. Herein, we performed the current study to probe into the regulatory mechanism of circ_0044516 in malignant behaviors of gastric cancer (GC) cells with the involvement of microRNA (miR)-149-5p/human antigen R (HuR) axis. Firstly, the expression levels of circ_0044516 in GC cell lines and normal gastric mucosal epithelial cells were determined by qRT-PCR, and GC cell lines with the highest expression of circ_0044516 were screened for further cell experiments. Subsequently, the biological functions of silenced circ_0044516 in GC were identified by CCK-8, colony formation, and transwell assays. Xenograft mouse models were established for in vivo verification. Furthermore, luciferase reporter, RIP, RNA pull-down assay and rescue experiments were performed to explore the sponge regulatory mechanism of circ_0044516. circ_0044516 was suggested to be highly expressed in GC cell lines, and circ_0044516 could promote GC cell proliferation, migration and invasion, as well as in vivo tumor growth. In addition, silenced circ-0044516 reversed the promotive roles in cell viability caused by overexpressed HuR. Furthermore, circ_0044516 mainly localized in the cytoplasm, which may act as a miR-149-5p sponge to modulate HuR expression, thereby playing an essential role in GC development. This study suggests that circ_0044516 may promote HuR expression through sponging miR-149-5p, thereby playing a part in GC progression.
环状 RNA(circRNAs)已成为一类多功能 RNA,但其在癌症发展中的功能知之甚少。在此,我们进行了当前的研究,以探究 circ_0044516 通过 microRNA(miR)-149-5p/人类抗原 R(HuR)轴在胃癌(GC)细胞恶性行为中的调控机制。首先,通过 qRT-PCR 测定 GC 细胞系和正常胃黏膜上皮细胞中 circ_0044516 的表达水平,并筛选出 circ_0044516 表达最高的 GC 细胞系进行进一步的细胞实验。随后,通过 CCK-8、集落形成和 Transwell 实验鉴定沉默 circ_0044516 在 GC 中的生物学功能。建立异种移植小鼠模型进行体内验证。此外,还进行了荧光素酶报告、RIP、RNA 下拉实验和挽救实验,以探索 circ_0044516 的海绵调控机制。提示 circ_0044516 在 GC 细胞系中高表达,circ_0044516 可促进 GC 细胞增殖、迁移和侵袭,以及体内肿瘤生长。此外,沉默 circ-0044516 逆转了过表达 HuR 引起的细胞活力促进作用。此外,circ_0044516 主要定位于细胞质,可能作为 miR-149-5p 的海绵,调节 HuR 表达,从而在 GC 发展中发挥重要作用。本研究表明,circ_0044516 可能通过海绵 miR-149-5p 促进 HuR 表达,从而在 GC 进展中发挥作用。