Growth Arrest and Apoptosis of the Human Hepatocellular Carcinoma Cell Line Bel-7402 Induced by Melittin

Abstract

Objective: The aim of this study was to investigate the cellular effects of melittin on the growth and apoptosis of human hepatocellular carcinoma (HCC) cells and to provide the molecular mechanism for potential application of a recombinant adenovirus carrying the melittin gene (Ad-rAFP-Mel) in the treatment of liver cancer. Methods: Human HCC cells (BEL-7402) were infected with Ad-rAFPMel at different times. In vitro cell growth was determined by MTT assay. Cellular apoptosis was evaluated quantitatively and qualitatively by phase-contrast microscopy, transmission electron microscopy, DNA ladder electrophoresis, TUNEL staining and flow cytometry. Results: Ad-rAFP-Mel infection had an inhibitory effect on the proliferation of BEL-7402 cells. The morphological changes of apoptosis were confirmed by microscopy and DNA electrophoresis. The ultrastructural characteristics of apoptotic cells, such as chromatin condensation and nuclear fragmentation, were also observed by electron microscopy in the Ad-rAFP-Mel-infected cells. Ad-rAFPMel infection markedly induced cellular apoptosis, and Fas expression on Bel-7402 cells infected by Ad-rAFPMel was up-regulated. Conclusion: The fact that melittin can induce apoptosis of the HCC cell line BEL-7402 leads us to consider adenovirus-mediated delivery of melittin as a promising approach for the treatment of HCC. However, the underlying mechanism needs to be further investigated.

High content analysis to determine cytotoxicity of the antimicrobial peptide, melittin and selected structural analogs

“…It increased IC in osteoblast-like cells [32], fibroblasts [33], hepatocytes [16,34] and mast cells [35]. In human osteoblasts, melittin increased IC levels only when calcium was present extracellularly and not from intracellular stores [32].…”

Bee venom suppresses PMA-mediated MMP-9 gene activation via JNK/p38 and NF-κB-dependent mechanisms

“…Bee venom consists of many biologically active enzymes, peptides and biogenic amines, such as melittin (the major active ingredient of bee venom), apamin, adolapin and mast cell-degranulating peptide (Park et al, 2004;Tu et al, 2008). Bee venom was reported to cause growth arrest or exhibited cytotoxic effects in hepatocellular carcinoma cells (Chu et al, 2007;Li et al, 2006). Also, proliferation of melanoma cells, activity of apoptotic enzyme (bcl-2 and caspase-3) in leukemic cells and ERK and Akt signaling pathway of renal cancer cells were all regulated and/or suppressed by bee venom (Moon et al, 2006;Son et al, 2007;Liu et al, 2002;Jang et al, 2003).…”

Experimental models of hepatocellular carcinoma: developments and evolution

“…Overexpression of cytokeratin 19 (CK19) correlates with HCC metastasis. The serum CK19 level correlated well with lung metastasis of the animal model, and the increased serum level of CK19 was also associated with the incidence of portal vein tumor embolus in patients with HCC (Ding et al 2004;Li et al 2006a, b). Metastasis is the result of cancer cell adaptation to a tissue microenvironment at a distance from the primary tumor (Albini 2008).…”