Fig. 2. ACSL6 promotes cell proliferation and migration in vitro and facilitates liver cancer growth and metastasis in vivo.

(A and B) Colony formation (A) and CCK-8 assays (B) of shNT or shACSL6 expressed MHCC97H and Huh7 cells. OD_{450 nm}, optical density at 450 nm. (C and D) Transwell (C) and wound healing assays (D) in MHCC97H and Huh7 cells expressing shNT or shACSL6. (E to G) Subcutaneous injection of shNT, shACSL6, and shACSL6 rescued with resistant ACSL6 wild-type (rACSL6 WT) or enzymatically dead (ED) MHCC97H cells into nude mice. Tumor volumes (E), tumor weights (F), and Ki-67 staining (G) in the xenograft model. (H to J) Subcutaneous injection of vector, ACSL6 WT–, or ED-overexpressing HepG2 cells into nude mice. Tumor volumes (H), tumor weights (I), and Ki-67 staining (J) in the xenograft model. (K to M) Assessment of the effect of high or low ACSL6 expression in tumors obtained from patients with liver cancer on PDX mouse models. Tumor volumes (K), tumor weights (L), and relative proliferative cells (M) in the PDX mouse model. (N and O) Representative images of hematoxylin and eosin (H&E) staining (N) and statistical analysis (O) of metastatic lung nodules from mice injected with shNT, shACSL6, and shACSL6 rescued with rACSL6 WT or ED MHCC97H cells via the tail vein. (P and Q) Representative images of H&E staining (P) and statistical analysis (Q) of metastatic lung nodules from mice injected with vector, ACSL6 WT–, or ED-overexpressed HepG2 cells via the tail vein. [(E) to (Q)] n = 5. *P < 0.05, **P < 0.01, and ***P < 0.001. Student’s t test [(A), (C), (D), (L), and (M)], two-way ANOVA [(B), (E), (H), and (K)], one-way ANOVA [(F), (G), (I), (J), (O), and (Q)].