[1]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].陆军军医大学学报(原第三军医大学学报),2010,32(09):930-933.
 Zeng Zhu,Long Jinhua.Hepatocellular carcinoma cells suppress mitochondrial function of dendritic cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):930-933.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第09期
页码:
930-933
栏目:
论著
出版日期:
2010-05-15

文章信息/Info

Title:
Hepatocellular carcinoma cells suppress mitochondrial function of dendritic cells in vitro
作者:
曾柱龙金华
贵阳医学院基础医学院生物技术教研室;北京大学基础医学院生物物理系血液流变学研究中心;贵州省肿瘤医院头颈肿瘤科
Author(s):
Zeng Zhu Long Jinhua
Department of Biotechnology, School of Basic Medical Science, Guiyang Medical College, Guiyang,  Guizhou Province, 550004; Hemorheological Research Center, Department of Biophysics, School of Basic Medical Science, Beijing University, Beijing, 100083; Department of Head and Neck Tumor, Tumor Hospital of Guizhou Province, Guiyang,  Guizhou Province, 550004, China
关键词:
树突状细胞肝癌细胞线粒体功能
Keywords:
dendritic cells hepatocellular carcinoma cells mitochondrial function
分类号:
R392.12;R730.23;R735.7
文献标志码:
A
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 necrosis factor -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)(P<0.05),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)(P<0.05, P<0.01),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)(P<0.01)。   结论   HCCs可能通过抑制DCs的线粒体功能来损伤其免疫功能。
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 DCs (imDCs). Maturation was induced by addition of 20 ng/ml rhTNF-α to imDCs for another 3 d of culture. Acquired imDCs and mDCs were respectively co-cultured with human HCCs for 48 h in Transwell chamber. These treated cells were investigated for mitochondrial membrane potential,   enzyme activity  and intracellular Ca2+ concentration ([Ca2+]in) in the cytoplasma by immune fluorescence and MTT assay. Those untreated cells served as  control.    Results   After co-cultured with HCCs, flow cytometry showed that the mitochondrial membrane potentials of imDCs and mDCs were respectively decreased from (659.991±17.052)and(473.741±11.676)to(482.681±7.935)and(407.189±5.051)(P<0.05) , [Ca2+]in of imDCs and mDCs were respectively increased from (427.983±3.358)and(232.587±3.815)to(517.308±0.249)and(413.062±2.981)(P<0.05, P<0.01). MTT assay showed that the enzyme activities of mitochondrions of imDCs and mDCs were respectively decreased from (0.764±0.089) and (0.708±0.019)to(0.291±0.019)and(0.218±0.019)(P<0.01).    Conclusion   HCCs might suppress immune function of DCs by inhibiting functional status of mitochondrion.

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更新日期/Last Update: 2010-05-07