Figure 2.

Stimulation of NK‐1R could enhance the proliferative, migrative and invaded effects in vitro. GBC‐SD cells (A) or NOZ cells (B) were seeded into 96‐well plates (6.0 × 10³ cells per well) and then treated with various concentrations of SP. The cell viability was determined using MTT assay at the indicated times. GBC‐SD cells (C) or NOZ cells (D), were treated with L703606 at the indicated concentrations for different times. The cell viability was measured using MTT assay. Effects of SP/NK‐1R system on clone formation ability in GBC‐SD and NOZ cells (E). The invasion of GBC‐SD and NOZ cells were examined by transwell assay. After pretreatment with L703606, cells were stimulated with SP, and the cells that invaded across the membrane of the transwell were stained with hematoxylin and eosin (F). GBC‐SD cells were scraped with a pipette tip and then treated with SP or L703606 for different times. The migrating cells were detected using a light microscope (G). NOZ cells were scraped and migration was determined by microscope (H). Data were shown as means ± SEM (n = 6). Statistical analysis was performed using one‐way ANOVA coupled with a post hoc test. Significant differences were indicated as *P < 0.05 versus Vehicle, **P < 0.01 versus Vehicle