Department of Thoracic Surgery, Cancer Center, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou 310014, China.
Zhejiang Provincial People's Hospital.
Crit Rev Eukaryot Gene Expr. 2024;34(4):13-23. doi: 10.1615/CritRevEukaryotGeneExpr.v34.i4.20.
Lung adenocarcinoma (LUAD) severely affects human health, and cisplatin (DDP) resistance is the main obstacle in LUAD treatment, the mechanism of which is unknown. Bioinformatics methods were utilized to predict expression and related pathways of AURKB in LUAD tissues, as well as the upstream regulated microRNAs. qRT-PCR assayed expression of AURKB and microRNA-486-5p. RIP and dual-luciferase experiments verified the binding and interaction between the two genes. CCK-8 was used to detect cell proliferation ability and IC50 values. Flow cytometry was utilized to assess the cell cycle. Comet assay and western blot tested DNA damage and γ-H2AX protein expression, respectively. In LUAD, AURKB was upregulated, but microRNA-486-5p was downregulated. The targeted relationship between the two was confirmed by RIP and dual-luciferase experiments. Cell experiments showed that AURKB knock-down inhibited cell proliferation, reduced IC50 values, induced cell cycle arrest, and caused DNA damage. The rescue experiment presented that high expression of microRNA-486-5p could weaken the impact of AURKB overexpression on LUAD cell behavior and DDP resistance. microRNA-486-5p regulated DNA damage to inhibit DDP resistance in LUAD by targeting AURKB, implying that microRNA-486-5p/AURKB axis may be a possible therapeutic target for DDP resistance in LUAD patients.
肺腺癌 (LUAD) 严重影响人类健康，而顺铂 (DDP) 耐药是 LUAD 治疗的主要障碍，其机制尚不清楚。本研究采用生物信息学方法预测 LUAD 组织中 AURKB 的表达及其相关通路，以及上游调控的 microRNA。qRT-PCR 检测 AURKB 和 microRNA-486-5p 的表达。RIP 和双荧光素酶实验验证了两个基因之间的结合和相互作用。CCK-8 检测细胞增殖能力和 IC50 值。流式细胞术检测细胞周期。彗星实验和 Western blot 分别检测 DNA 损伤和 γ-H2AX 蛋白表达。在 LUAD 中，AURKB 上调，而 microRNA-486-5p 下调。RIP 和双荧光素酶实验证实了两者之间的靶向关系。细胞实验表明，AURKB 敲低抑制细胞增殖，降低 IC50 值，诱导细胞周期停滞，并导致 DNA 损伤。挽救实验表明，microRNA-486-5p 的高表达可以减弱 AURKB 过表达对 LUAD 细胞行为和 DDP 耐药性的影响。microRNA-486-5p 通过靶向 AURKB 调节 DNA 损伤来抑制 LUAD 中的 DDP 耐药性，这表明 microRNA-486-5p/AURKB 轴可能是 LUAD 患者 DDP 耐药的潜在治疗靶点。
