Anti-human hepatocellular carcinoma effects of tumor necrosis factor-related apoptosis-inducing ligand in vivo and in vitro

AIM: To investigate the effect of over-expression of Bcl-2 protein on Trail protein-induced apoptosis in human hepatoma cells, and the cytotoxicity of Trail protein on human hepatoma cells in vitro and in vivo. METHODS: The T rail gene was cloned and expressed in E coli. The cytotoxicity of the recom binant Trail protein was assayed on human hepatoma cells in vitro and in vi vo. The cell viability was assessed by trypan blue exclusion. The stable hu man hepatoma cells clone in which Bcl-2 protein over-expressed was establis hed by transfecting eukaryotic expression plasmid pcDNA3-Bcl-2 into BEL-74- 04 human hepatoma cells, and was selected with G418 400 mg (.) L-1. RESULTS : The recombinant Trail protein actively killed human hepatoma cells tested in this study such as BEL-7404, BEL-7402, and SMMC-7721. Over-expression o f Bcl-2 protein could inhibit apoptosis induced by Trail in BEL-7404 human hepatoma cells in vitro. It was obvious that the purified recombinant Trail protein could inhibit tumor formation of BEL-7404. human hepatoma cells in nude mice. CONCLUSION: The recombinant Trail protein could kill human hepa toma cells in vitro and in vivo. Overexpression of Bcl-2 protein could inhi bit Trail-induced apoptosis in BEL-7404. human hepatoma cells. The results suggested that Trail might be a potential agent for the liver cancer therap y.