Figure 6

Brachyury silencing reduces the tumorigenicity and metastasis of ACCS-M GFP cells. A. ACCS-M GFP derivatives transfected with shRNAs (untreated control, ACCS-M GFP [a, d, g]; Brachyury shRNA, M sh.Bra [b, e, h]; SOX2 shRNA, M sh.SOX2 [c, f, i]) were injected into the tongues of nude mice and examined to detect tumors in tongues (a–f) and spontaneous metastases in submandibular lymph nodes (g–i). Observations with the naked eye (a–c) and the excitation of GFP (d–i) are shown. Sites of lymph node metastasis are labeled with arrowheads. Note that GFP enables the detection of micro-metastasis in the lymph nodes. B. The primary tumor volumes were measured weekly, calculated as the length × width × thickness, and mice were sacrificed when the primary tumor volume reached 100 mm³. Tumor growth curves for ACCS derivatives ACCS-M GFP (diamond), sh.Bra (square), sh.SOX2 (triangle), and sh cont. (control shRNA, circle) are shown. Experiments were performed in 6 mice for each ACCS derivative, and the values were averaged. Bars indicate the standard deviation. Data significances between ACCS-M GFP and other ACCS derivatives were analyzed by Student’s t-test. *P < 0.05, **P < 0.01.