抗人死亡受体5单链抗体ZF1对鼠H22肝癌细胞的作用分析

Analysis of single-chain antibody against death receptor 5 (ZF1) on murine H22 hepatocellular carcinoma cell

目的:研究抗人死亡受体5单链抗体ZF1对H22肝癌细胞体内外抑制增殖作用。方法:MTT法检测ZF1对H22肝癌细胞体外杀伤作用,流式细胞术检测ZF1诱导H22的凋亡率,建立H22移植瘤模型,随机分为PBS组、ZF1组、EPI组和ZF1/EPI联合组四组。观察肿瘤生长和小鼠体重变化情况。治疗13天后分离肿瘤组织,进行HE染色检查、TUNNEL法检测细胞凋亡。结果:体外实验显示:ZF1可抑制H22细胞的增殖,呈剂量依赖性,抑制率最高为84.5%。体内实验结果显示单独应用ZF1或联合应用ZF1/EPI时,肿瘤增长受到明显抑制。HE染色和TUNNEL分析结果表明ZF1可有效诱导肝癌肿瘤凋亡,ZF1/EPI联合组效果更明显,而对正常肝细胞无毒性。结论:单链抗体ZF1具有良好抑制H22细胞增殖的作用。ZF1和EPI联合应用效果更明显。

Objective：The purpose of this study is to evaluate the effects of a single-chain antibody against death receptor 5 （ZF1） on tumor growth and survival in murine H22 hepatocellular carcinoma tumor model.Methods：Killing effect of ZF1 on H22 cells was analyzed by MTT assay in vitro. The apoptosis rate of H22 cells induced by ZF1 was detected using Flow Cytometry assay. The transplanted model of H22 tumor was developed in mice. The mice were randomly divided into four groups, PBS group, ZF1 group, EPI group and combined treatment group of ZF1/EPI. Tumor growth and body weight changes were observed. After treatment over 13 days, the tumor tissue for HE staining and TUNNEL assay was performed to detect apoptosis.Results：The results showed that ZF1 could inhibit growth of H22 cells in a dose dependent manner. The growth inhibition rate was up to 84.5%. The results showed that ZF1 alone or in combination with ZF1/EPI, the tumor growth was significantly inhibited. HE staining and TUNNEL analysis showed that ZF1 could effectively induce apoptosis of tumor cells without toxic effects, especially in ZF1/EPI combined treatment group.Conclusion：It is showed that ZF1 induces a good inhibition on the proliferation of H22 cell, especially in combined treatment group of ZF1/EPI.