Department of Thoracic Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei Province 050000, China.
Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei Province 050000, China.
J Immunol Res. 2022 Jul 15;2022:4996980. doi: 10.1155/2022/4996980. eCollection 2022.
BACKGROUND/AIM: Non-small-cell lung cancer (NSCLC) is the principal agent of cancer deaths globally. The goal of this study was to determine how circular RNA_0000518 (circ_0000518) regulates tumor progression. . circ_0000518 was selected as a study target involved in NSCLC from GEO (Gene Expression Omnibus) database. circ_0000518 level was gauged by qRT-PCR. It was confirmed as circRNA by actinomycin D inhibition and RNase R assay. Subcellular localization of circ_0000518 was identified by FISH. Cell function was determined by CCK-8, Transwell, and western blot. Glutamine metabolic factors were detected by ELISA. The target regulation relationship between genes was clarified by dual-luciferase reporter assay. In vivo models were established to evaluate the impact of circ_0000518 on tumor growth. Immunohistochemical staining for Ki67, vimentin, and E-cadherin was used to detect cell proliferation and metastasis, respectively.
circ_0000518 expression was enhanced in NSCLC. si-circ_0000518 inhibited cell proliferation, invasion, and glutamine metabolism. circ_0000518 functioned as a molecular sponge for miR-330-3p, and inhibition of miR-330-3p in cells markedly reversed circ_0000518 interference-mediated antitumor effects. miR-330-3p interacted with 3'-UTR of SLC1A5. miR-330-3p inhibitor-mediated protumor effect was remarkably reversed in cells after the knockdown of SLC1A5. circ_0000518 knockdown reduced glutamine, glutamate, and -KG by targeting miR-330-3p. Intertumoral injection of circ_0000518 shRNA adeno-associated virus effectively halted xenograft tumor growth.
The current study revealed that circ_0000518 may have a prooncogenic function in the formation and progression of NSCLC, which might be achieved through moderating the miR-330-3p/SLC1A5 axis.
背景/目的：非小细胞肺癌（NSCLC）是全球癌症死亡的主要原因。本研究的目的是确定环状 RNA_0000518（circ_0000518）如何调节肿瘤进展。 。从 GEO（基因表达综合数据库）数据库中选择环状 RNA_0000518 作为 NSCLC 的研究靶点。通过 qRT-PCR 测量 circ_0000518 水平。通过放线菌素 D 抑制和 RNase R 测定证实为 circRNA。通过 FISH 鉴定 circ_0000518 的亚细胞定位。通过 CCK-8、Transwell 和 Western blot 测定细胞功能。通过 ELISA 检测谷氨酰胺代谢因子。通过双荧光素酶报告基因测定阐明基因之间的靶标调节关系。建立体内模型以评估 circ_0000518 对肿瘤生长的影响。通过免疫组化染色 Ki67、波形蛋白和 E-钙黏蛋白分别检测细胞增殖和转移。
circ_0000518 在 NSCLC 中表达增强。si-circ_0000518 抑制细胞增殖、侵袭和谷氨酰胺代谢。circ_0000518 作为 miR-330-3p 的分子海绵发挥作用，并且细胞中 miR-330-3p 的抑制显著逆转了 circ_0000518 干扰介导的抗肿瘤作用。miR-330-3p 与 SLC1A5 的 3'-UTR 相互作用。在 SLC1A5 敲低后，细胞中 miR-330-3p 抑制剂介导的促肿瘤作用显著逆转。circ_0000518 通过靶向 miR-330-3p 减少谷氨酰胺、谷氨酸和 -KG。肿瘤内注射 circ_0000518 shRNA 腺相关病毒有效地阻止了异种移植肿瘤的生长。
本研究表明，circ_0000518 可能在 NSCLC 的形成和进展中具有致癌功能，这可能是通过调节 miR-330-3p/SLC1A5 轴实现的。
