Li Peiyuan, Xiao Weisheng
Department of Gastroenterology, the First Affiliated Hospital of Hengyang Medical College, University of South China, China.
Department of Gastroenterology, the First Affiliated Hospital of Hengyang Medical College, University of South China, China.
Toxicol In Vitro. 2022 Dec;85:105454. doi: 10.1016/j.tiv.2022.105454. Epub 2022 Aug 12.
Circular RNAs (circRNAs) have been reported to have roles in the carcinogenesis of gastric cancer (GC). Circ_0005758 was discovered to be decreased in GC, here, the detailed functions and molecular mechanism of circ_0005758 in GC progression were investigated. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to measure the levels of genes and proteins. The biological functions of circ_0005758 on GC progression were investigated by using in vitro assays, including 5-ethynyl-2'-deoxyuridine (EDU), transwell, tube formation and flow cytometry, and in vivo murine xenograft model. The binding between miR-1229-3p and circ_0005758 or GCNT4 (Glucosaminyl (N-Acetyl) Transferase 4) was confirmed using dual-luciferase reporter assay and pull-down assay. Circ_0005758 expression was decreased in GC tissues and cells, re-expression of circ_0005758 induced apoptosis and suppressed proliferation, migration, invasion and angiogenesis in GC cells in vitro, and impeded xenograft tumor growth in nude mice. Mechanistically, circ_0005758 sequestered miR-1229-3p to release GCNT4 expression, indicating the circ_0005758/miR-1229-3p/GCNT4 competing endogenous RNA (ceRNA) network GC cells. Besides, an increased miR-1229-3p level and a decreased GCNT4 expression were observed in GC. Rescue experiments demonstrated that miR-1229-3p up-regulation or GCNT4 down-regulation attenuated the anticancer effects of circ_0005758 re-expression on GC cells. Circ_0005758 acts as a tumor suppressor to impede gastric cancer progression via miR-1229-3p/GCNT4 axis, implying that therapeutic targeting of circ_0005758 may better to prevent gastric cancer.
据报道，环状RNA（circRNAs）在胃癌（GC）的致癌过程中发挥作用。研究发现circ_0005758在GC中表达降低，在此，对circ_0005758在GC进展中的详细功能和分子机制进行了研究。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测基因和蛋白质水平。通过体外实验，包括5-乙炔基-2'-脱氧尿苷（EDU）、Transwell、管腔形成和流式细胞术，以及体内小鼠异种移植模型，研究circ_0005758对GC进展的生物学功能。使用双荧光素酶报告基因检测和下拉实验证实了miR-1229-3p与circ_0005758或GCNT4（氨基葡萄糖基（N-乙酰）转移酶4）之间的结合。circ_0005758在GC组织和细胞中表达降低，circ_0005758的重新表达诱导GC细胞凋亡，并抑制其体外增殖、迁移、侵袭和血管生成，同时抑制裸鼠体内异种移植肿瘤的生长。机制上，circ_0005758通过结合miR-1229-3p来释放GCNT4的表达，表明在GC细胞中存在circ_0005758/miR-1229-3p/GCNT4竞争性内源RNA（ceRNA）网络。此外，在GC中观察到miR-1229-3p水平升高和GCNT4表达降低。挽救实验表明，miR-1229-3p上调或GCNT4下调减弱了circ_0005758重新表达对GC细胞的抗癌作用。circ_0005758作为一种肿瘤抑制因子，通过miR-1229-3p/GCNT4轴阻碍胃癌进展，这意味着靶向circ_0005758进行治疗可能更有利于预防胃癌。
