Moreover, treatment with NOZ and GBC-SD cells with rh-TSP-4 alone also facilitated their invasion and cancer stem-like properties (Additional file 5: Fig. S2E-S2G), and the presence of rh-TSP-4 prominently promoted the expression of EMT and cancer stemness markers in NOZ and GBC-SD cells in a dose-dependent manner, as confirmed by western blot analysis (Additional file 5: Fig. S2H). This evidence concerns the gene THBS4 and cancer.