Department of Thoracic Surgery, Jianhu Hospital Affiliated to Nantong University, Yancheng, China.
Information Center, Jianhu Hospital Affiliated to Nantong University, Yancheng, China.
Bioengineered. 2021 Dec;12(1):2326-2340. doi: 10.1080/21655979.2021.1935064.
Non-small cell lung cancer (NSCLC) is a common malignant tumor, with high morbidity and mortality. Circular RNA (circRNA) circ_0003028 was reported to be upregulated in NSCLC. This study is designed to explore the role and mechanism of circ_0003028 on NSCLC progression. In this work, circ_0003028, microRNA-1298-5p (miR-1298-5p), and glutamic oxaloacetic transaminase 2 (GOT2) level were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The localization of circ_0003028 was analyzed by subcellular fractionation assay. Cell proliferation, colony number, cell cycle progression, apoptosis, migration, invasion, and angiogenesis were measured by Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, transwell, and tube formation assays. Protein levels of Beclin1, light chain 3 (LC3)-II/LC3-I, GOT2, proliferating cell nuclear antigen (PCNA) were examined by western blot assay. The binding relationship between miR-1298-5p and circ_0003028 or GOT2 was predicted by circular RNA Interactome or starbase and then verified by dual-luciferase reporter, RNA Immunoprecipitation (RIP), and RNA pull-down assays. The biological role of circ_0003028 on NSCLC tumor growth was examined by the xenograft tumor model . We reported that circ_0003028 and GOT2 were upregulated, and miR-1298-5p was decreased in NSCLC tissues and cells. Moreover, circ_0003028 knockdown curbed cell proliferative ability, migration, invasion, angiogenesis, and facilitate apoptosis and autophagy in NSCLC cells . Mechanical analysis discovered that circ_0003028 regulated GOT2 expression by sponging miR-1298-5p. Circ_0003028 silencing hindered the cell growth of NSCLC . Taken together, circ_0003028 knockdown could suppress NSCLC progression partly by regulating the miR-1298-5p/GOT2 axis, providing an underlying therapeutic target for NSCLC.
非小细胞肺癌（NSCLC）是一种常见的恶性肿瘤，发病率和死亡率都很高。Circ_0003028 已被报道在 NSCLC 中上调。本研究旨在探讨 circ_0003028 对 NSCLC 进展的作用和机制。在这项工作中，通过实时定量聚合酶链反应（RT-qPCR）检测 circ_0003028、microRNA-1298-5p（miR-1298-5p）和谷草转氨酶 2（GOT2）的水平。通过亚细胞分离测定分析 circ_0003028 的定位。通过细胞计数试剂盒-8（CCK-8）、集落形成、流式细胞术、Transwell 和管形成测定法测量细胞增殖、集落数、细胞周期进程、细胞凋亡、迁移、侵袭和血管生成。通过 Western blot 测定法检测 Beclin1、轻链 3（LC3）-II/LC3-I、GOT2、增殖细胞核抗原（PCNA）的蛋白水平。通过 circRNA Interactome 或 starbase 预测 miR-1298-5p 与 circ_0003028 或 GOT2 的结合关系，然后通过双荧光素酶报告、RNA 免疫沉淀（RIP）和 RNA 下拉测定验证。通过异种移植肿瘤模型检测 circ_0003028 对 NSCLC 肿瘤生长的生物学作用。我们报道 circ_0003028 和 GOT2 在 NSCLC 组织和细胞中上调，而 miR-1298-5p 下调。此外，circ_0003028 敲低抑制 NSCLC 细胞的增殖能力、迁移、侵袭、血管生成，并促进细胞凋亡和自噬。机械分析发现 circ_0003028 通过海绵吸附 miR-1298-5p 调节 GOT2 表达。circ_0003028 沉默抑制 NSCLC 细胞的生长。总之，circ_0003028 敲低通过调节 miR-1298-5p/GOT2 轴可能抑制 NSCLC 的进展，为 NSCLC 提供了一个潜在的治疗靶点。
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