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肝癌细胞对树突状细胞线粒体功能的抑制作用
引用本文:曾柱,龙金华.肝癌细胞对树突状细胞线粒体功能的抑制作用[J].第三军医大学学报,2010,32(9).
作者姓名:曾柱  龙金华
作者单位:1. 550004 贵阳,贵阳医学院基础医学院生物技术教研室;100083,北京,北京大学基础医学院生物物理系血液流变学研究中心
2. 贵州省肿瘤医院头颈肿瘤科,贵阳,550004
基金项目:贵州省省长基金,贵州省自然科学基金 
摘    要:目的探索肝癌细胞(hepatocellular carcinoma cells,HCCs)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,以进一步理解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(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(recombinant human tumor necrosisfactor-α,rhTNF-α)将imDCs诱导为成熟DCs(mature DCs,mDCs),分别将imDCs和mDCs与肝癌细胞在Transwell中共培养48h,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比色法研究HCCs对DCs的线粒体膜电位、酶活力和胞内钙离子浓度的影响。结果与HCCs共培养后,流式细胞仪的检测结果显示:imDCs和mDCs的线粒体膜电位分别从(659.991±17.052)和(473.741±11.676)下降到(482.681±7.935)和(407.189±5.051)(P0.05),imDCs和mDCs胞内钙离子浓度分别从(427.983±3.358)和(232.587±3.815)增加到(517.308±0.249)和(413.062±2.981)(P0.05,P0.01),MTT比色法的检测结果显示imDCs和mDCs的酶活力分别从(0.764±0.089)和(0.708±0.019)下降到(0.291±0.019)和(0.218±0.019)(P0.01)。结论HCCs可能通过抑制DCs的线粒体功能来损伤其免疫功能。

关 键 词:树突状细胞  肝癌细胞  线粒体功能

Hepatocellular carcinoma cells suppress mitochondrial function of dendritic cells in vitro
Zeng Zhu,Long Jinhua.Hepatocellular carcinoma cells suppress mitochondrial function of dendritic cells in vitro[J].Acta Academiae Medicinae Militaris Tertiae,2010,32(9).
Authors:Zeng Zhu  Long Jinhua
Institution:Zeng Zhu1,2,Long Jinhua3 (1Department of Biotechnology,School of Basic Medical Science,Guiyang Medical College,Guiyang,Guizhou Province,550004,2Hemorheological Research Center,Department of Biophysics,Beijing University,Beijing,100083,3Department of Head and Neck Tumor,Tumor Hospital of Guizhou Province,China)
Abstract:Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the functional status of mitochondria in dendritic cells (DCs) in order to investigate the mechanisms of tumor immune escape.Methods CD14+ monocytes,which were isolated with immune magnetic beads from fresh peripheral blood of healthy human,were then cultured in RPMI1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DC...
Keywords:dendritic cells  hepatocellular carcinoma cells  mitochondrial function  
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