Fig. 4

Effect of ghrelin and In1-ghrelin overexpression on PCa pathophysiological processes. a. Cell proliferation of empty (mock), ghrelin and In1-ghrelin vectors stably transfected PC-3 and VCaP cell lines for 24-48 h (n ≥ 3); b. Cell migration of mock, ghrelin and In1-ghrelin stably transfected PC-3 cell line after 12 h by wound-healing assay (n ≥ 3). Representative images showing the migration capacity of PC-3 cells transfected with mock, ghrelin and In1-ghelin are also indicated; c. phospho-ERK and phospho-AKT basal activation in PC-3 stably transfected cells (n ≥ 3). Blots are representative of one cell passage with three technical replicates; d. Growth rate of subcutaneously inoculated mock, ghrelin and In1-ghrelin-transfected PC-3-derived tumors in nude mice (n = 5) followed up to 12 weeks after inoculation. Statistical significance was evaluated by two-way ANOVA (***p < 0.001 indicate differences between In1-ghrelin and mock, while #p < 0.001 indicates differences between In1-ghrelin and ghrelin stably transfected cells); e. % of necrosis in xenografted PC-3-derived tumors. Representative images of hematoxylin–eosin (H/E) staining are depicted. f. N° of mitosis/10 fields in xenografted PC-3-derived tumors. Data represent mean ± SEM (n ≥ 3). Asterisks (***p < 0.001; **p < 0.01; *p < 0.05) indicate values that significantly differ from the mock control