hsa_circ_0017639 的表达通过海绵吸附 miR-224-5p 和上调 USP3 促进胃癌的增殖和转移。

Hsa_circ_0017639 expression promotes gastric cancer proliferation and metastasis by sponging miR-224-5p and upregulating USP3.

机构信息

Department of Gastroenterology, The First Affiliated Hospital of Soochow University, 188 Shizi Road, Suzhou 215006, PR China; Department of Gastroenterology, Shanghai Gongli Hospital, the Second Military Medical University, 219 Miaopu Road, Pudong New Area, Shanghai 200135, PR China.

Shanghai Key Laboratory of Molecular Imaging, Shanghai University of Medicine and Health Sciences, Shanghai 201318, PR China.

出版信息

Gene. 2020 Aug 5;750:144753. doi: 10.1016/j.gene.2020.144753. Epub 2020 May 4.

DOI:10.1016/j.gene.2020.144753

PMID:32376451

Abstract

Gastric cancer (GC) is a common malignant tumor having poor prognosis globally. Circular RNA (circRNA) is a circular endogenous RNA generated by special selective splicing that occurs in various traits. Studies show that hsa_circ_0017639 is abnormally expressed and involved in tumorigenesis. Nevertheless, the hsa_circ_0017639 role in GC is unknown. This study detected hsa_circ_0017639 expression in a GC cell line using RT-qPCR. Subcellular localization of hsa_circ_0017639 was verified via FISH. We assessed correlations amongst miRNA, hsa_circ_0017639 and relative protein levels using luciferase reporter assays and RNA pulldown assays. The data illustrated that in hsa_circ_assays, expression was enhanced in GC cell. Downregulation of hsa_circ_0017639 decreased GC cell proliferation and migration in in vitro and in vivo experiments. RNA pulldown and RT-qPCR analysis verified that hsa_circ_0017639 sponged miR-224-5p. Bioinformatic and luciferase reporter assays validated that miR-224-5p and USP3 were downstream targets of hsa_circ_0017639. Upregulation of USP3 or downregulation of miR-224-5p restored proliferation and migration by MKN-28 and MGC-803 cells after hsa_circ_0017639 silencing. Upregulation of USP3 restored MKN-28 and MGC-803 cell proliferation and migration after overexpression of miR-224-5p. Our collective findings advised that hsa_circ_0017639 takes part in GC progression through regulating the miR-224-5p/USP3 axis, highlighting its potential as an effective GC therapeutic target.

摘要

胃癌（GC）是一种常见的恶性肿瘤，全球预后较差。环状 RNA（circRNA）是一种通过特殊的选择性剪接产生的环状内源性 RNA，存在于各种特征中。研究表明 hsa_circ_0017639 表达异常，并参与肿瘤发生。然而，hsa_circ_0017639 在 GC 中的作用尚不清楚。本研究通过 RT-qPCR 检测了 GC 细胞系中 hsa_circ_0017639 的表达。通过 FISH 验证 hsa_circ_0017639 的亚细胞定位。我们通过荧光素酶报告基因检测和 RNA 下拉实验评估了 miRNA、hsa_circ_0017639 和相对蛋白水平之间的相关性。结果表明，在 hsa_circ_0017639 表达增强的 GC 细胞中。在体外和体内实验中，下调 hsa_circ_0017639 可降低 GC 细胞的增殖和迁移。RNA 下拉和 RT-qPCR 分析证实 hsa_circ_0017639 可吸附 miR-224-5p。生物信息学和荧光素酶报告基因检测验证了 miR-224-5p 和 USP3 是 hsa_circ_0017639 的下游靶点。上调 USP3 或下调 miR-224-5p 可恢复 hsa_circ_0017639 沉默后 MKN-28 和 MGC-803 细胞的增殖和迁移。上调 miR-224-5p 可恢复 MKN-28 和 MGC-803 细胞的增殖和迁移。我们的研究结果表明，hsa_circ_0017639 通过调节 miR-224-5p/USP3 轴参与 GC 进展，提示其作为一种有效的 GC 治疗靶点的潜力。