Clinical Medicine College, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Department of Clinical Medicine, North Sichuan Medical College, Nanchong, China.
Gut Liver. 2023 Mar 15;17(2):267-279. doi: 10.5009/gnl210542. Epub 2022 Sep 23.
BACKGROUND/AIMS: We aimed to investigate the role and working mechanism of Homo sapiens circular RNA_0003602 (hsa_circ_0003602) in colorectal cancer (CRC) development.
The expression of circ_0003602, miR-149-5p, and solute carrier family 38 member 1 (SLC38A1) was detected by quantitative real-time polymerase chain reaction. RNase R assays were conducted to determine the characteristics of circ_0003602. CCK-8 assays, flow cytometry analysis, transwell invasion assays, wound healing assays and tube formation assays were employed to evaluate cell viability, apoptosis, invasion, migration, and angiogenesis. All protein levels were examined by Western blot or immunohistochemistry assay. The glutamine metabolism was monitored by corresponding glutamine, α-ketoglutarate and glutamate assay kits. Dual-luciferase reporter assay was utilized to confirm the targeted combination between miR-149-5p and circ_0003602 or SLC38A1. A xenograft tumor model was established to analyze the role of circ_0003602 in CRC tumor growth in vivo.
Circ_0003602 was upregulated in CRC tissues and cell lines. Circ_0003602 silencing suppressed CRC cell viability, migration, invasion, angiogenesis, and glutaminolysis; induced cell apoptosis in vitro; and blocked tumor growth in vivo. Moreover, circ_0003602 directly interacted with miR-149-5p to negatively regulate its expression, and circ_0003602 knockdown suppressed the malignant behaviors of CRC cells largely by upregulating miR-149-5p. MiR-149-5p directly bound to the 3' untranslated region of SLC38A1 to induce its degradation, and miR-149-5p overexpression reduced the malignant potential of CRC cells largely by downregulating SLC38A1. Circ_0003602 positively regulated SLC38A1 expression by sponging miR-149-5p in CRC cells.
Circ_0003602 knockdown impedes CRC development by targeting the miR-149-5p/SLC38A1 axis, which provides a novel theoretical basis and new insights for CRC treatment.
背景/目的：本研究旨在探究人类环状 RNA_0003602（hsa_circ_0003602）在结直肠癌（CRC）发展中的作用和工作机制。
采用实时定量聚合酶链反应检测 circ_0003602、miR-149-5p 和溶质载体家族 38 成员 1（SLC38A1）的表达。采用 RNase R 分析检测 circ_0003602 的特征。通过 CCK-8 检测、流式细胞术分析、Transwell 侵袭实验、划痕愈合实验和管形成实验评估细胞活力、凋亡、侵袭、迁移和血管生成。通过 Western blot 或免疫组化检测所有蛋白水平。通过相应的谷氨酰胺、α-酮戊二酸和谷氨酸检测试剂盒监测谷氨酰胺代谢。利用双荧光素酶报告基因实验证实 miR-149-5p 与 circ_0003602 或 SLC38A1 之间的靶向结合。建立异种移植肿瘤模型，分析 circ_0003602 在体内 CRC 肿瘤生长中的作用。
circ_0003602 在 CRC 组织和细胞系中上调。circ_0003602 沉默抑制 CRC 细胞活力、迁移、侵袭、血管生成和谷氨酰胺分解；体外诱导细胞凋亡；体内阻断肿瘤生长。此外，circ_0003602 与 miR-149-5p 直接相互作用，负调控其表达，circ_0003602 沉默主要通过上调 miR-149-5p 抑制 CRC 细胞的恶性行为。miR-149-5p 直接结合 SLC38A1 的 3'非翻译区诱导其降解，miR-149-5p 过表达通过下调 SLC38A1 降低 CRC 细胞的恶性潜能。circ_0003602 通过海绵吸附 miR-149-5p 在 CRC 细胞中正向调控 SLC38A1 的表达。
circ_0003602 通过靶向 miR-149-5p/SLC38A1 轴抑制 CRC 发展，为 CRC 治疗提供了新的理论依据和新的见解。
