目的:肺癌是全球死亡的主要原因之一。最近的研究表明,环状RNA在多种癌症中失调,但是肺癌的机制仍然不清楚。在我们的工作中,我们探讨了circLDLRAD3在肺癌中的作用机制。
方法:通过实时定量聚合酶链反应(RT-qPCR)检测肺癌中circLDLRAD3,microRNA-497-5p(miR-497-5p)和血小板型PFK(PFKP)的丰度。同时,通过蛋白质印迹定量PFKP水平.细胞计数试剂盒-8(CCK-8),5-乙炔基-2'-脱氧尿苷(EdU)测定,transwell分析,伤口愈合试验,流式细胞术,westernblot,进行免疫组织化学(IHC)测定和糖酵解代谢分析以进行功能分析。此外,miR-497-5p与circLDLRAD3或FKPF之间的相互作用通过双荧光素酶报告基因和RNA免疫沉淀(RIP)检测.最终,体内实验用于测量circLDLRAD3的作用。
结果:circLDLRAD3和PFKP水平升高。沉默circLDLRAD3抑制细胞活力,扩散,迁移,肺癌细胞的侵袭和糖酵解代谢及促进细胞凋亡。在机制上,circLDLRAD3作为miR-497-5p海绵调节PFKP水平。MiR-497-5p通过抑制PFKP抑制肺癌的进展。此外,circLDLRAD3敲低也抑制体内肿瘤生长。
结论:CircLDLRAD3通过调节miR-497-5p增加PFKP的表达促进肺癌的发展,这也为肺癌治疗提供了潜在的靶向治疗。
OBJECTIVE: Lung cancer is one of the leading causes of death worldwide. Recent studies have shown that circular RNAs are dysregulated in a variety of cancers, but the mechanism in lung cancer is still indistinct. In our work, we explored the action mechanism of circLDLRAD3 in lung cancer.
METHODS: The abundance of circLDLRAD3, microRNA-497-5p (miR-497-5p) and platelet-type PFK (
PFKP) was measured by real-time quantitative polymerase chain reaction (RT-qPCR) in lung cancer. Meanwhile, the level of
PFKP was quantified by western blot. Cell counting kit-8 (CCK-8), 5-Ethynyl-2\'-deoxyuridine (EdU) assay, transwell assay, wound healing assay, flow cytometry, western blot, immunohistochemical (IHC) assay and glycolysis metabolism analysis were performed for functional analyses. Furthermore, the interplay between miR-497-5p and circLDLRAD3 or FKPF was detected by the dual-luciferase reporter and RNA Immunoprecipitation (RIP) assays. Eventually, the in vivo experiments were applied to measure the role of circLDLRAD3.
RESULTS: The levels of circLDLRAD3 and
PFKP were increased. Silencing circLDLRAD3 inhibited cell viability, proliferation, migration, invasion and glycolysis metabolism and promoted cell apoptosis in lung cancer cells. In mechanism, circLDLRAD3 regulated
PFKP level as a miR-497-5p sponge. MiR-497-5p suppressed the progression of lung cancer by inhibiting
PFKP. In addition, circLDLRAD3 knockdown also inhibited tumor growth in vivo.
CONCLUSIONS: CircLDLRAD3 promoted the development of lung cancer through increasing PFKP expression by regulating miR-497-5p, which also provided a potential targeted therapy for lung cancer treatment.