肝癌细胞对树突状细胞线粒体功能的影响.docVIP

肝癌细胞对树突状细胞线粒体功能的影响 曾 柱1，2，龙金华3 （550004 贵阳，贵阳医学院生物技术教研室1；100083北京，北京大学基础医学院生物物理系2；550004 贵阳，贵州省肿瘤医院3） 【摘 要】 目的 探讨肝癌细胞分泌的可溶性细胞因子营造的微环境对树突状细胞（Dendritic cells，DCs）线粒体功能状态的影响，以探索肿瘤的免疫逃逸机制。方法 用免疫磁珠从人外周血分离CD14＋单核细胞，加入粒-巨噬细胞集落刺激因子（Recombinant human granulocyte-macrophage CSF, rhGM-CSF）、白介素4（Recombinant human interleukins 4, IL-4）将单核细胞诱导分化为未成熟DCs（immature DCs，imDCs），利用肿瘤坏死因子α（Tumor necrosis factor-α, TNF-α）将imDCs诱导为成熟DCs（mature DCs, mDCs），分别将imDCs和mDCs与肝癌细胞（Hepatocellular carcinoma cells, HCC）在Transwell中共培养48 h，正常培养和撤生长因子培养的DCs作为对照，利用免疫荧光和MTT比色法等方法研究HCC对DCs的胞内钙离子浓度、线粒体膜电位和酶活力的影响。结果 与正常培养和撤生长因子培养的DCs相比，与HCC共培养DCs的胞内钙离子浓度增加、线粒体膜电位和酶活力受到了显著的抑制。结论 HCC能够通过抑制DCs的线粒体功能损伤其免疫功能。 【关键词】 树突状细胞；肝癌细胞；线粒体功能 【中图分类号】 Q6-3 【文献标志码】 A 【基金项目】 贵州省省长基金（黔省专字2009-79）；贵州省自然科学基金（黔省合J字2008-2274） 【通信作者】曾 柱，E-mail：zengzhu100@ The Effect of Hepatocellular Carcinoma Cells on the Mitochondrial Function of Dendritic Cells Zeng Zhu1,2, Long Jinhua3 （1Department of Biotechnology, Guiyang Medical College, Guiyang, Guizhou, 550004；2Department of Biophysics, Health Science Center, Peking University, Beijing, 100083,；3Tumor Hospital of Guizhou, Guiyang, Guizhou, 550004, China） 【Abstract】：Objective The progression of tumor generation is accompanied by a marked suppression of the effect units of immune system. DCs are most powerful antigen presenting cells as now known and play significant roles in the initiations of innate and acquired immune response. This study focused on the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCC) on the functions of mitochondrions in DCs, and exploring the mechanisms of tumor immune escape. Methods: The monocytes were then cultured in RPMI 1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 days to develop into immature DCs (imDCs). Maturation was induced by addition of 20 ng/ml rhTNF-α to imDCs for another 3 days of culture. Acquired imDCs and mDCs were respectively co-cultured with human HC