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2016-11-29 发布于河北
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Cytotoxic activity induced by dendritic cell vaccine plus gemcitabine to the hepatocellular carcinom.doc

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Cytotoxic activity induced by dendritic cell vaccine plus gemcitabine to the hepatocellular carcinom

树突状细胞瘤苗联合吉西他滨对肝癌干细胞的杀伤效应王建，任雷，郭源，滕木俭（山东大学附属千佛山医院肝胆外科，济南250014）【摘要】：目的探讨负载CD133+肝癌细胞抗原的树突状细胞（DC）诱导的免疫治疗联合吉西他滨（GEM）在体外对肝癌干细胞的杀伤效应。方法取对数生长期的人肝癌细胞系Huh-7以CD133作为分子标志进行流式分选,得到肝癌干细胞。将人外周血单个核细胞（PBMC）在体外诱导分化为树突状细胞（DC）。DC负载Huh-7细胞和CD133+细胞抗原后与T细胞共育得到特异性细胞毒性T细胞，将CD133+细胞作为靶细胞进行细胞毒性试验。实验组按处理因素分为：1.GEM组；2.CD133+-CTL组； 3. GEM+CD133+-CTL组；4.Huh7-CTL组；5.GEM+Huh7-CTL组。CCK-8法检测杀伤率，然后比较各组间差异。结果对CD133+细胞的杀伤效应以GEM+CD133+-CTL组最强，与其他各组比较差异均有统计学意义（P＜0.05）。而单独就DC瘤苗杀伤率而言，CD133+-CTL组要高于Huh7-CTL组（P＜0.05）。结论 CD133+肝癌干细胞裂解产物致敏的DC瘤苗联合化疗药物可以有效杀伤肝癌干细胞，进而可能降低肝癌术后和肝癌肝移植后的转移和复发率，为临床治疗提供新的思路。关键词：树突状细胞瘤苗；吉西他滨；肝癌干细胞中图分类号: R735.7 文献标志码: A Cytotoxic activity induced by dendritic cell vaccine plus gemcitabine to the hepatocellular carcinoma cancer stem cell WANG Jian, REN Lei, GUO Yuan, TENG Mu-jian (Department of Hepatobiliary Surgery, Qianfoshan Hospital Affiliated to Shandong University, Jinan 250014, China) [Abstract] Objective To investigate the cytotoxicity of gemcitabine(GEM) combined with immunotherapy mediated by dendritic cells(DCs) loaded with CD133+ liver cancer cell antigens to hepatocellular carcinoma cancer stem cells(HCC CSCs) in vitro. Methods The HCC CSCs were sorted from Huh-7 cells in logarithmicgrowth phase by flow cytometery via the CD133 molecular mark. Human peripheral blood mononuclear cells(PBMCs) were induced into dendritic cells in vitro. Then theDCs were sensitized by Huh-7 and CD133+ cancer cell antigens. After antigen loading-DCs were co-cultured with T cells，specific cytotoxic T lymphocytes were obtained. CD133+ cells were taken as target cells for cytotoxicity tests. HCC CSCs were divided into five groups according to treatment factors: GEM group, CD133+-CTL group, GEM+CD133+-CTL group, Huh7-CTL group and GEM+Huh7-CTL group. The cytotoxic effects were assessed by CCK-8 assay. Then variances were compared and analyzed. Results The GEM+CD133+-CTL group have the strongest cytotoxic effect to target cells, when compared with other groups. And the differences were st