Department of Respiratory Medicine, Hangzhou Dingqiao Hospital, Hangzhou, China.
Department of Emergency, Hangzhou Dingqiao Hospital, Hangzhou, China.
Cancer Biother Radiopharm. 2020 Nov;35(9):673-681. doi: 10.1089/cbr.2019.3258. Epub 2020 Mar 31.
Studies showed that miR-532-5p suppresses proliferation and induces apoptosis of lung cancer (LC) cells; its role in LC is not fully understood. Therefore, this research aimed to reveal the effect and mechanism of miR-532-5p on migration and invasion of LC cells. The transfection efficiencies of miR-532-5p mimic, inhibitor, and overexpressed were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The relationships between miR-532-5p and in A549 and SBC-5 cells were predicted by targetScan and verified by dual-luciferase reporter assay, Western blot, and qRT-PCR. Migration and invasion of cells transfected with miR-532-5p mimic, inhibitor, and were determined by scratch test and transwell assay, respectively. The levels of epithelial-to-mesenchymal transition (EMT)-related proteins (E-cadherin (E-Cad)), N-catenin (N-Cad), and vimentin) in cells were measured by Western blot. MiR-532-5p mimic suppressed migration and invasion, while miR-532-5p inhibitor promoted migration and invasion of cells. was a downstream target of miR-532-5p and both its protein and mRNA expressions were inhibited by miR-532-5p mimic, but promoted by miR-532-5p inhibitor. promoted migration, invasion, and EMT process, and such effects of were reversed by miR-532-5p mimic. MiR-532-5p functioned as a cancer suppressor by negatively regulating in LC cells, pointing to a potential protective mechanism of miR-532-5p to LC patients.
研究表明 miR-532-5p 可抑制肺癌(LC)细胞增殖并诱导其凋亡,但 miR-532-5p 在 LC 中的作用尚不完全清楚。因此,本研究旨在揭示 miR-532-5p 对 LC 细胞迁移和侵袭的影响及作用机制。通过实时定量聚合酶链反应(qRT-PCR)测定 miR-532-5p 模拟物、抑制剂和过表达载体的转染效率。通过靶标扫描预测 miR-532-5p 与 A549 和 SBC-5 细胞中 的关系,并通过双荧光素酶报告基因检测、Western blot 和 qRT-PCR 进行验证。划痕实验和 Transwell 实验分别检测转染 miR-532-5p 模拟物、抑制剂和 载体的细胞的迁移和侵袭能力。Western blot 检测细胞中上皮间质转化(EMT)相关蛋白(E-钙黏蛋白(E-Cad)、N-钙黏蛋白(N-Cad)和波形蛋白)的水平。miR-532-5p 模拟物抑制细胞迁移和侵袭,而 miR-532-5p 抑制剂则促进细胞迁移和侵袭。 是 miR-532-5p 的下游靶基因,miR-532-5p 模拟物抑制 蛋白和 mRNA 的表达,而 miR-532-5p 抑制剂则促进其表达。 促进迁移、侵袭和 EMT 过程,而 miR-532-5p 模拟物则逆转了 的这些作用。miR-532-5p 通过负向调控 LC 细胞中的 发挥抑癌作用,提示 miR-532-5p 对 LC 患者具有潜在的保护机制。