龙金华,曾柱
LONG Jinhua1,ZENG Zhu2(1.the Affiliated Cancer Hospital of Guiyang Medical College

• 1:贵阳医学院附属肿瘤医院
• 2:贵阳医学院生物技术教研室

摘要(Abstract)：

目的:探索血管内皮生长因子(vascular endothelial growth factor,VEGF)对成熟树突状细胞(maturedendritic cells,mDCs)蛋白质表达谱的影响,以进一步理解肿瘤的免疫逃逸机制,为改善基于DCs的抗肿瘤免疫的临床治疗效率寻找新的线索。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),VEGF作用于mDCs,用基于质谱的蛋白质组技术研究VEGF对mDCs的蛋白质表达谱的影响。结果:VEGF可上调或下调mDCs的细胞骨架及其相关蛋白、迁移相关蛋白、免疫功能相关蛋白和抗氧化相关蛋白的表达。结论:VEGF可能通过重组mDCs的细胞骨架来损伤其运动能力和免疫学功能,这对进一步理解DCs的生物学功能和肿瘤的免疫逃逸机制具有重要意义。
Objective: To investigate the effect of vascular endothelial growth factor(VEGF) on protein expression profile of mature dendritic cells(mDCs),and so as to further understanding of the immune escape mechanism of tumor and looking for the new clue for improvement of clinical efficiency of DCs-based immune therapy against cancer.Methods: CD14+ monocytes were isolated from peripheral blood by immunomagnetic beads.The monocytes were then cultured with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF) and recombinant human interleukins 4(rhIL-4) for 7 days to develop into immature DCs(imDCs).Tumor necrosis factor-α(rhTNF-α) was used to induce imDCs into mCDs.The mDCs were treated with VEGF.The protein expression profiles of mCDs were investigated by mass spectrometry-based proteomics technique.Results: Some cytoskeletal proteins and their related proteins,migration-associated proteins,immune function-related proteins and antioxidation-associated proteins in mCDs were up or down-regulated by VEGF.Conclusions: The motility and immune function of mDCs might be impaired by VEGF through cytoskeletal reorganization,which is significant for further understanding of biological function of mDCs and immune escape mechanism of tumor.

关键词(KeyWords)： 树突细胞;成熟树突状细胞;血管内皮生长因子;蛋白质组学
dendritic cells;mature dendritic cells,vascular endothelial growth factor;proteomics

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(11162003);; 教育部科学技术研究重点项目(210196);; 贵州省优秀青年科技人才支持计划(2011-24);; 贵州省社会发展科技攻关项目(黔科合SY字[2011]3065);; 贵州省省长专项基金(黔省专合字[2009]-79);; 贵州省科学技术基金(黔科合J字2008-2274);; 贵阳市科学技术项目(2010-筑科农合同字第1-社-12)

作者(Author): 龙金华,曾柱
LONG Jinhua1,ZENG Zhu2(1.the Affiliated Cancer Hospital of Guiyang Medical College

DOI: 10.19367/j.cnki.1000-2707.2012.04.003

参考文献(References)：

• [1]Steinman RM,Banchereau J.Taking dendritic cells intomedicine[J].Nature,2007(449):419-426.
• [2]Steinman,RM.Decisions about dendritic cells:past,pres-ent,and future[J].Annu Rev Immunol,2012(30):1-22.
• [3]Schreiber RD,Old LJ,Smyth MJ.Cancer immunoedit-ing:integrating immunity's roles in cancer suppressionand promotion[J].Science,2011(331):1565-1570.
• [4]Rabinovich GA,Gabrilovich D,Sotomayor EM.Immuno-supressive strategies that are mediated by tumor cells[J].Annu Rev Immunol,2007(25):267-296.
• [5]Johnson BF,Clay TM,Hobeika AC,et al.Vascular en-dothelial growth factor and immunosuppression in cancer:current knowledge and potential for new therapy[J].Ex-pert Opin Biol Ther,2007(7):449-460.
• [6]Masoumi MS,Amini A,Morris DL,et al.Significance ofvascular endothelial growth factor in growth and peritonealdissemination of ovarian cancer[J].Cancer MetastasisRev,2012(31):143-62.
• [7]Steinman RM.Dendritic cells in vivo:a key target for a newvaccine science[J].Immunity,2008(29):319-324.
• [8]Zeng Z,Xu XF,Zhang YY,et al.Tumor-derived factorsimpaired motility and immune functions of dendritic cellsthrough derangement of biophysical characteristics and re-organization of cytoskeleton[J].Cell Motilty and Cy-toskeleton,2007(64):186-98.
• [9]Zeng Z,Yao WJ,Xu XF,et al.Hepatocellular carcino-ma cells deteriorate the biophysical properties of dendriticcells[J].Cell Biochem Biophys,2009(55):33-43.
• [10]Xu XF,Zeng Z,Wang XW,et al.Biomechanical altera-tions of dendritic cells by co-culturing with K562 CMLcells and their potential role in immune escape[J].J Bi-omechanics,2010(43):2339-2347.
• [11]Mimura K,Kono K,Takahashi A,et al.Vascular endo-thelial growth factor inhibits the function of human maturedendritic cells mediated by VEGF receptor-2[J].CancerImmunol Immunother,2007(56):761-70.
• [12]Freeman SA,Lei V,Dang-Lawson M,et al.Cofilin-me-diated F-actin severing is regulated by the Rap GTPaseand controls the cytoskeletal dynamics that drive lympho-cyte spreading and BCR microcluster formation[J].J Im-munol,2011(187):5887-5900.
• [13]Ishaq M,Lin BR,Bosche M,et al.LIM kinase 1-de-pendent cofilin 1 pathway and actin dynamics mediate nu-clear retinoid receptor function in T lymphocytes[J].BMC Mol Biol,2011(12):41-53.
• [14]Yun SP,Ryu JM,Jang MW,et al.Interaction of profi-lin-1 and F-actin via a beta-arrestin-1/JNK signalingpathway involved in prostaglandin E(2)-induced humanmesenchymal stem cells migration and proliferation[J].JCell Physiol,2011(226):559-571.
• [15]Machaidze G,Sokoll A,Shimada A,et al.Actin fila-ment bundling and different nucleating effects of mouseDiaphanous-related formin FH2 domains on actin/ADFand actin/cofilin complexes[J].J Mol Biol,2010(403):529-545.
• [16]Wang WA,Groenendyk J,Michalak M.Calreticulin sig-naling in health and disease[J].Int J Biochem Cell Bi-ol,2012(44):842-846.
• [17]Choi HJ,Choi WS,Park JY,et al.SIGN-R1,a C-typelectin,binds to Bip/GRP78 and this interaction mediatesthe regurgitation of T-cell-independent type 2 antigen dex-tran through the endoplasmic reticulum[J].Immunobiol-ogy,2011(216):437-446.
• [18]Fang H,Wu Y,Huang X,et al.Toll-like receptor 4(TLR4)is essential for Hsp70-like protein 1(HSP70L1)to activate dendritic cells and induce Th1response[J].J Biol Chem,2011(286):30393-30400.
• [19]Fricke I.Vascular endothelial growth factor-trap over-comes defects in dendritic cell differentiation but does notimprove antigen-specific immune responses[J].ClinCancer Res,2007(13):4840-4848.
• [20]Xu J,Li T,Wu H,et al.Role of thioredoxin in lung dis-ease[J].Pulm Pharmacol Ther,2012(25):154-162.
• [21]Branco V,Canário J,Lu J,et al.Mercury and seleniuminteraction in vivo:effects on thioredoxin reductase andglutathione peroxidase[J].Free Radic Biol Med,2012(52):781-793.
• [22]Verma-Gaur J,Hauser J,Grundstrm T.Negative feedbackregulation of antigen receptors through calmodulin inhibitionof E2A[J].J Immunol,2012(188):6175-83.
• [23]Smith AL,Ganesh L,Leung K,et al.Leukocyte-specif-ic protein 1 interacts with DC-SIGN and mediates trans-port of HIV to the proteasome in dendritic cells[J].JExp Med,2007(204):421-30.
• [24]Pan X,Mao X,Cheng T,et al.Macrophage migrationinhibitory factor:a regulator of MMP13 and inflammationin titanium particles-stimulated air pouch in vivo[J].Mol Cell Biochem,2011(357):313-321.
• [25]Heinrichs D,Knauel M,Offermanns C,et al.Macro-phage migration inhibitory factor(MIF)exerts antifibroticeffects in experimental liver fibrosis via CD74[J].ProcNatl Acad Sci USA,2011(108):17444-17449.
• [26]Wittkowski H,Kuemmerle-Deschner JB,Austermann J,et al.MRP8 and MRP14,phagocyte-specific danger sig-nals,are sensitive biomarkers of disease activity incryopyrin-associated periodic syndromes[J].Ann RheumDis,2011(70):2075-2081.
• [27]Maiseyeu A,Badgeley MA,Kampfrath T,et al.In vivotargeting of inflammation-associated myeloid-related pro-tein 8/14 via gadolinium immunonanoparticles[J].Ar-terioscler Thromb Vasc Biol,2012(32):962-970.
• [28]Garcia-Mata R,Boulter E,Burridge K.The'invisiblehand:regulation of Rho GTPases by RhoGDIs[J].NatRev Mol Cell Biol,2011(22):493-504.

文章评论(Comment)：