Fig. 5: miR-596-3p suppresses the metastatic ability of PC9-BrM by inhibiting YAP1-mediated MMP-2 expression.

A A schematic diagram of in vitro BBB model. B PC9-BrM cells transfected with the miR-596-3p mimics, or both miR-596-3p mimics and YAP1 were cultured on the up chamber pre-coated with normal human astrocytes (NHA) and Human brain Microvascular endothelial cells (HBMEC) on both sides, and the migrated cells was measured after 24 h. C PC9 and PC9-BrM cells with the transfected with YAP1 were added on the up chamber and the number of invaded cells was measured after 24 h. D Transwell assay for PC9-BrM cells transfected with the miR-596-3p, or both miR-596-3p and YAP1 was performed. E CCK-8 assay for PC9-BrM cells transfected with the miR-596-3p, or both miR-596-3p and YAP1 was conducted. F MMP-2 expression was detected in PC9-BrM cells and PC9 cells that were transfected with miR-596-3p mimics or miR-inhibitor+YAP1 by ELISA. G MMP-2 mRNA expression was explored in PC9 cells that were transfected with miR-596-3p inhibitor/miR-inhibitor NC or pcDNA3.1-YAP1/vector or miR-inhibitor+YAP1 by qRT-PCR. H Kaplan–Meier analysis for brain metastasis-free survival of NSCLC patients using a combined GEO databases. Patients were divided into two groups based on the expression status of MMP-2 in their primary tumors. Data were presented as the means ± standard error of the mean of at least three independent experiments. ^*P < 0.05, ^**P < 0.01 and ^***P < 0.001.