Department of Cardiothoracic Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Department of Cardiothoracic Surgery, Lishui City People's Hospital, the Sixth Affiliated Hospital of Wenzhou Medical University, Lishui, China.
Technol Cancer Res Treat. 2021 Jan-Dec;20:1533033820985868. doi: 10.1177/1533033820985868.
We aimed to investigate the mechanism of the regulatory axis of miR-196b/AQP4 underlying the invasion and migration of lung adenocarcinoma (LUAD) cells.
LUAD miRNA and mRNA expression profiles were downloaded from TCGA database and then differential analysis was used to identify the target miRNA. Target gene for the miRNA was obtained via prediction using 3 bioinformatics databases and intersection with the differentially expressed mRNAs searched from TCGA-LUAD. Then, qRT-PCR and western blot were used to validate the expression of miR-196b and AQP4. Dual-luciferase reporter assay was performed to confirm the targeting relationship between miR-196b and AQP4. Transwell assay was used to investigate the migration and invasion of LUAD cells.
MiR-196b was screened out by differential and survival analyses, and the downstream target gene AQP4 was identified. In LUAD, miR-196b was highly expressed while AQP4 was poorly expressed. Besides, overexpression of miR-196b promoted cell invasion and migration, while overexpression of AQP4 had negative effects. Moreover, the results of the dual-luciferase reporter assay suggested that AQP4 was a direct target of miR-196b. In addition, we also found that overexpressing AQP4 could suppress the promotive effect of miR-196b on cancer cell invasion and migration.
MiR-196b promotes the invasion and migration of LUAD cells by down-regulating AQP4, which helps us find new molecular targeted therapies for LUAD.
本研究旨在探讨 miR-196b/AQP4 调控轴在肺腺癌（LUAD）细胞侵袭和迁移中的作用机制。
从 TCGA 数据库中下载 LUAD 的 miRNA 和 mRNA 表达谱，然后进行差异分析以鉴定靶 miRNA。通过 3 个生物信息学数据库进行预测，获得 miRNA 的靶基因，并与从 TCGA-LUAD 中搜索到的差异表达 mRNA 进行交集。然后，采用 qRT-PCR 和 Western blot 验证 miR-196b 和 AQP4 的表达。通过双荧光素酶报告基因实验证实 miR-196b 与 AQP4 之间的靶向关系。采用 Transwell 实验研究 LUAD 细胞的迁移和侵袭。
通过差异分析和生存分析筛选出 miR-196b，确定其下游靶基因 AQP4。在 LUAD 中，miR-196b 呈高表达，AQP4 呈低表达。此外，过表达 miR-196b 促进细胞侵袭和迁移，而过表达 AQP4 则产生抑制作用。双荧光素酶报告基因实验结果表明，AQP4 是 miR-196b 的直接靶基因。此外，我们还发现过表达 AQP4 可以抑制 miR-196b 对癌细胞侵袭和迁移的促进作用。
miR-196b 通过下调 AQP4 促进 LUAD 细胞的侵袭和迁移，这有助于我们为 LUAD 找到新的分子靶向治疗方法。
