Fig. 5. Functional effects of ALCAM downregulation in GCTB cells.
a Knockdown of ALCAM inhibited the proliferative rate of GCTB28 cells. Independent experiments were repeated three times. Data were presented as mean ± s.d. (***p < 0.001; **p < 0.01; *p < 0.05). b Knockdown of ALCAM reduced the size and number of spheres in GCTB28 cells. (Scale bar = 20 µm). c, d Knockdown of ALCAM in GCTB28 cells resulted in significant decrease in (c) invasive and (d) migratory capacities, respectively. Ten high power fields were selected for comparison. Data were presented as mean ± s.d. (***p < 0.001; **p < 0.01; *p < 0.05). (Scale bar = 50 µm). e, f ALCAM knockdown (sh ALCAM) GCTB cells exhibited reduced tumor-forming incidence when compared with NTC cells. ALCAM knockdown (sh ALCAM) GCTB28 cells and NTC cells with different gradient (1 × 103 cells, 5 × 103 cells, 2.5 × 104 cells and 1 × 105 cells; n = 5 in each gradient) were injected subcutaneously into flank of nude mice. See also TableS4. e Representative images of sh ALCAM and NTC cells induced tumor formation at 8 weeks after 1 × 105 cells injection. White arrow indicates tibia destruction. f H&E staining of paraffin-embedded tumor tissue from mice after orthotopic injection of NTC cells. The histological features of tumor xenograft induced by the NTC cells are comparable to bone tissue by the sh ALCAM cells (Scale bar = 20 µm)