Fig. 5

SNS-032 mitigates migration and invasion of human UM cells. a After treatment with SNS-032 for the indicated time periods, wound-healing scratch assay in 92.1 (top) and Omm2.3 (bottom) cells was performed. Scale bar: 200 μm. b SNS-032 suppressed the migration of UM cells examined by transwell assays. Migrated cells were counted in five randomly selected fields. **, P < 0.01, Student’s t test. Scale bar: 200 μm. c SNS-032 suppressed the invasion of UM cells examined by matrigel invasion assays. Invaded cells were counted in five randomly selected fields. Data represent mean ± SD. **, P < 0.01, Student’s t test. Scale bar: 200 μm. d SNS-032 inhibited the protein levels of MMP9. 92.1 and Omm2.3 cells were treated with various concentrations of SNS-032 for 48 h, MMP9 and MMP2 levels were detected by Western blot analysis. e SNS-032 suppressed the gene transcription of MMP9. The mRNA levels of MMP9 were measured after 24 h exposure to SNS-032 by qRT-PCR. *, P < 0.05; **, P < 0.01; ***, P < 0.001, one-way ANOVA, post hoc intergroup comparisons, Tukey’s test. f Analysis of a set of UM patients (n = 77) from TCGA database showed that higher MMP9 expression predicted shorter overall survival (P < 0.001). g and h YAP was critical in SNS-032-mediated decrease of MMP9. 92.1 cells were transfected with Vector and YAP (PIP-HA-YAP) constructs (left) or transduced with lentiviral control (Scramble) and shRNA against YAP (right), then treated with SNS-032 for 24 h. Overexpression or knockdown of YAP was confirmed. The protein and mRNA levels of MMP9 were examined by Western blot and qRT-PCR analysis, respectively. *, P < 0.05; **, P < 0.01, Student’s t test. i SNS-032 reduced YAP binding to the gene promoter of MMP9. Mel270 cells were treated with SNS-032 (0.5 μM) for 24 h, the recruitment of YAP on the gene promoter of MMP9 was measured by ChIP assay. IgG was used as negative control. ***, P < 0.001, Student’s t test