肿瘤坏死因子相关凋亡诱导配体的体内体外抗人肝细胞癌的作用

目的：研究Bcl-2蛋白的过表达对Trail诱导肝癌细胞凋亡的影响，以及Trail蛋白体内和体外对人肝癌细胞的杀伤作用。方法：克隆Trail基因并在大肠杆菌中表达Trail重组蛋白。检测Trail重组蛋白在体内和体外对肿瘤的杀伤作用。用台盼蓝拒染法检测细胞的凋亡率。将真核细胞表达质粒pcDNA3-Bcl-2转染到人肝癌细胞BEL－7404细胞中，并通过G418 400mg/L筛选得到Bcl-2蛋白稳定表达的细胞株。结果：重组蛋白Trail能够有效地杀死人肝癌细胞（包括BEL－7404，SMMC－7721，和BEL－7402等细胞）。在体外，过量表达的Bcl-2蛋白能抑制Trail重组蛋白对人肝癌细胞BEL－7404的杀伤作用。重组蛋白Trail能够有效地抑制人肝癌细胞BEL－7404在裸鼠中形成实体瘤。结论：在体内和体外，Trail重组蛋白能够杀死肝癌细胞，Bcl-2蛋白过表达可以抑制Trail重组蛋白对肝癌细胞BEL－7404的杀伤作用。Trail蛋白是一种潜在的肝癌治疗剂。

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 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.