Anti-human hepatocellular carcinoma effects of tumor necrosis factor-related apoptosis-inducing ligand in vitro & in vivo
Abstract
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 Trail gene was cloned and expressed in E coli. The cytotoxicity of the recombinant Trail protein was assayed on human hepatoma cells in vitro and in vivo. The cell viability was assessed by trypan blue exclusion. The stable human hepatoma cells clone in which Bcl-2 protein over-expressed was established by transfecting eukaryotic expression plasmid pcDNA3-Bcl-2 into BEL-7404 human hepatoma cells, and was selected with G418 400 mg/L.
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 of 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 hepatoma cells in vitro and in vivo. Over-expression of Bcl-2 protein could inhibit Trail-induced apoptosis in BEL-7404 human hepatoma cells. The results suggested that Trail might be a potential agent for the liver cancer therapy.
Keywords:
Methods: The Trail gene was cloned and expressed in E coli. The cytotoxicity of the recombinant Trail protein was assayed on human hepatoma cells in vitro and in vivo. The cell viability was assessed by trypan blue exclusion. The stable human hepatoma cells clone in which Bcl-2 protein over-expressed was established by transfecting eukaryotic expression plasmid pcDNA3-Bcl-2 into BEL-7404 human hepatoma cells, and was selected with G418 400 mg/L.
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 of 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 hepatoma cells in vitro and in vivo. Over-expression of Bcl-2 protein could inhibit Trail-induced apoptosis in BEL-7404 human hepatoma cells. The results suggested that Trail might be a potential agent for the liver cancer therapy.