背景:MiR-21-5p是一种高表达的microRNA,在各种促进癌症的过程中起着重要作用,包括锚定独立生长,入侵,迁移转移,和肺癌的耐药性。研究表明,miR-21-5p可能通过促进上皮-间质转化(EMT)来促进这些过程。Ras同源基因家族成员B(RhoB),一个被miR-21-5p下调的基因,也与肺癌中的EMT有关。然而,miR-21-5p/RhoB轴在肺腺癌EMT调节中的作用尚不清楚.在这项研究中,我们旨在研究miR-21-5p/RhoB轴在EMT中的调节作用以及相关的体外功能特征,如迁移,入侵,顺铂耐药,和肿瘤球体的形成。
结果:用miR-21-5p抑制剂转染A549细胞,RhoBsiRNA,及其相应的阴性对照。伤口愈合,Transwell入侵,甲基噻唑四唑(MTT),和球体形成测定也进行了评估迁移,入侵,顺铂耐药,和A549细胞的锚定非依赖性生长。RT-qPCR用于确定EMT标志物的mRNA表达水平。MiR-21-5p敲低抑制迁移,入侵,顺铂耐药,和球体形成,同时上调E-cadherin和下调Slug。此外,RhoB沉默恢复了A549细胞的EMT和相关的体外功能特征。
结论:敲低miR-21-5p通过上调RhoB抑制EMT和相关的体外功能特征,提示miR-21-5p可能通过下调RhoB促进EMT。
BACKGROUND: MiR-21-5p is a highly expressed microRNA that plays an important role in various cancer-promoting processes, including anchorage-independent growth, invasion, migration metastasis, and drug resistance in lung cancer. Studies indicate that miR-21-5p may contribute to these processes by promoting epithelial-mesenchymal transition (EMT). Ras homolog gene family member B (RhoB), a gene downregulated by miR-21-5p, has also been linked to EMT in lung cancer. However, the role of the miR-21-5p/RhoB axis in EMT regulation in lung adenocarcinoma remains unclear. In this study, we aimed to investigate the regulatory role of the miR-21-5p/RhoB axis in EMT and related in vitro functional characteristics such as migration, invasion, cisplatin resistance, and the formation of tumor spheroids.
RESULTS: A549 cells were transfected with the miR-21-5p inhibitor, RhoB siRNA, and their corresponding negative controls. Wound healing, transwell invasion, Methyl thiazole tetrazolium (MTT), and sphere formation assays were also performed to evaluate the migration, invasion, cisplatin resistance, and anchorage-independent growth of A549 cells. RT-qPCR was used to determine the mRNA expression levels of EMT markers. MiR-21-5p knockdown inhibited migration, invasion, cisplatin resistance, and sphere formation while upregulating E-cadherin and downregulating Slug. Furthermore, RhoB silencing restored EMT and related in vitro functional characteristics in A549 cells.
CONCLUSIONS: Knockdown of miR-21-5p inhibits EMT and related in vitro functional characteristics by upregulating RhoB, suggesting that miR-21-5p may promote EMT through downregulation of RhoB.