Department of Cardio-Thoracic Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, No. 365 Renming East Road, Wucheng District, Jinhua, 321000, China.
Department of Traditional Chinese Medicine, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua, 321000, China.
Biochem Genet. 2022 Dec;60(6):1865-1880. doi: 10.1007/s10528-022-10191-x. Epub 2022 Feb 4.
We aimed to elucidate binding of microRNA-9-5p and STARD13 in lung adenocarcinoma (LUAD) cells and discuss their impact on malignant progression of LUAD, so as to provide evidence for identifying new therapeutic targets for LUAD. Bioinformatics analysis was introduced for analysis of differentially expressed miRNAs in LUAD tissue, and potential downstream target gene was predicted with TargetScan and other databases. MicroRNA-9-5p and STARD13 mRNA levels at cellular level was analyzed with qRT-PCR assay. Lipofectamine 2000 was applied for cell transfection. Proliferation, migration and invasion of LUAD cells were assayed with CCK-8, wound healing and Transwell assays, respectively. Protein expression of STARD13 was assessed with western blot. Binding of microRNA-9-5p and STARD13 was identified with dual-luciferase assay. Compared with normal human bronchial cells, microRNA-9-5p level in LUAD cells was noticeably increased, and STARD13 level was noticeably decreased. MicroRNA-9-5p could significantly promote malignant progression of LUAD cells, while forced STARD13 level markedly repress malignant progression of LUAD cells. Dual-luciferase gene assay showed that microRNA-9-5p had a direct targeting relationship with STARD13, and it was also found that microRNA-9-5p enhanced malignant behaviors of LUAD cells through modulating STARD13. STARD13 was a target of microRNA-9-5p in LUAD. MicroRNA-9-5p fostered malignant behaviors of LUAD cells by targeting STARD13. Therefore, microRNA-9-5p may become a new target for LUAD, and microRNA-9-5p inhibition may be a new treatment method.
我们旨在阐明 microRNA-9-5p 和 STARD13 在肺腺癌 (LUAD) 细胞中的结合，并探讨它们对 LUAD 恶性进展的影响，为寻找 LUAD 的新治疗靶点提供依据。采用生物信息学分析方法分析 LUAD 组织中差异表达的 miRNAs，利用 TargetScan 等数据库预测潜在的下游靶基因。采用 qRT-PCR 检测细胞水平 microRNA-9-5p 和 STARD13mRNA 水平。采用 Lipofectamine 2000 进行细胞转染。采用 CCK-8 法、划痕愈合实验和 Transwell 实验分别检测 LUAD 细胞的增殖、迁移和侵袭能力。采用 Western blot 检测 STARD13 蛋白表达。采用双荧光素酶报告基因实验鉴定 microRNA-9-5p 与 STARD13 的结合。与正常人支气管细胞相比，LUAD 细胞中 microRNA-9-5p 水平明显升高，STARD13 水平明显降低。microRNA-9-5p 可显著促进 LUAD 细胞的恶性进展，而强制 STARD13 水平则明显抑制 LUAD 细胞的恶性进展。双荧光素酶基因实验表明，microRNA-9-5p 与 STARD13 具有直接的靶向关系，还发现 microRNA-9-5p 通过调节 STARD13 增强 LUAD 细胞的恶性行为。STARD13 是 LUAD 中 microRNA-9-5p 的靶基因。microRNA-9-5p 通过靶向 STARD13 促进 LUAD 细胞的恶性行为。因此，microRNA-9-5p 可能成为 LUAD 的新靶点，抑制 microRNA-9-5p 可能成为 LUAD 的新治疗方法。
