Fig. 4

HJ-4 suppresses angiogenesis and tumor growth in colon cancer models. Colon cancer spheroids derived from HCT116 and SW480 cells were implanted into the vascular-rich region of the CAM. HJ-4 (8, 16, and 32 µM) or the positive control Piperine (32 µM) was topically applied to the tumor area daily for 7 consecutive days. Tumors were then excised, weighed, and vascular-like structures were recorded. (A) Representative images of vascular-like structures formed after treatment with HJ-4 (8, 16, 32 µM) or Piperine (32 µM). (B–C) Quantitative analysis of vascular-like structure formation in HCT116 (B) and SW480 (C) cells. HJ-4 significantly inhibited angiogenesis in a dose-dependent manner compared to the control group. Data are presented as mean ± SD (n = 3). (D–E) Representative images of xenograft tumors derived from HCT116 (D) and SW480 (E) cells treated with HJ-4 or Piperine. (F–G) Quantitative analysis of tumor weights in HCT116 (F) and SW480 (G) xenograft models. HJ-4 significantly reduced tumor weight in a dose-dependent manner and demonstrated greater efficacy than Piperine at 32 µM. Data are presented as mean ± SD (n = 6). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001.