Antitumor effect of the antimicrobial peptide GLI13-8 derived from domain of the avian β-defensin-4

We previously reported that GLI13-8, one of cationic antimicrobial peptides from linear avian β-defensin-4 (RL38) analogs, exhibited high antimicrobial activities against both Gram-negative and Gram-positive bacteria. In the present study, we reported the in vitro cytotoxicity of GLI13-8 using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results showed that the cytotoxicity of GLI13-8 in three human carcinoma cells (HepG2, SGC7901, and A375) was in a dose-dependent manner. When the concentration of GLI13-8 is <128 μM, it had no toxicity towards the normal human fibroblasts (MRC-5). The Annexin-V-FITC/PI staining assay, the Hoechst 33258/PI staining assay, the permeability of fluorescein macromolecules and scanning electron microscope assays, mitochondrial membrane potential assay, caspases-3 and poly ADP-ribose polymerase (PARP) assays have been carried out. Results indicated that apoptosis was induced by GLI13-8 in HepG2 cells, and demonstrated that GLI13-8 induced loss of mitochondrial membrane potential, disruption of HepG2 cell membranes, and activation of caspase-3 and PARP. These findings suggested that GLI13-8 may be an effective agent for HepG2 cells.