Myristoyl-CM4 Exhibits Direct Anticancer Activity and Immune Modulation in Hepatocellular Carcinoma: Evidence from In Vitro and Mouse Model Studies.

Hepatocellular carcinoma (HCC) is a major clinical challenge due to limited treatment options, More therapy candidates with confirmed anticancer effects are urgently needed. Antimicrobial peptide myristoyl-CM4 exhibits effective anticancer activity against leukemia and breast cancer cells. However, its therapeutic potential in HCC remains unexplored. The objective of the present study was to evaluate the anticancer activity of myristoyl-CM4 against cultured HCC cells and HCC xenograft tumors in mice. Cell viability, apoptosis, proliferation, epithelial-mesenchymal transition, migration, and invasion were assessed using standard assays. Mechanistic studies focused on its effects on macrophages utilized western blotting, immunofluorescence staining, and immunohistochemistry assays in HCC/macrophage co-culture models. The study results showed that myristoyl-CM4 induced apoptosis in HCC cells by targeting mitochondria. It also inhibited HCC cell migration and invasion in both HCC monoculture and HCC/macrophage co-culture systems. Notably, myristoyl-CM4 also promoted M1 macrophage polarization and suppressed M2 polarization in co-culture models both in vitro and in vivo. It also demonstrated effective antitumor activity both in PLC-PRF-5 xenograft and PLC-PRF-5/macrophage co-xenograft mouse models. Collectively, these findings highlighted the therapeutic potential of myristoyl-CM4 in HCC treatment.