张启芳;吴昌学;禹文峰;官志忠;柏华;
ZHANG Qifang;WU Changxue;YU Wenfeng;GUAN Zhizhong;BAI Hua;Key Laboratory of Molecular Biology,Guizhou Medical University;Medical Experimental Center,Third Affiliated Hospital of Guizhou Medical University;

• 1:贵州医科大学分子生物学重点实验室
• 2:贵州医科大学第三附属医院医学实验中心

摘要(Abstract)：

目的:研究放疗联合抑制信号转导与转录激活因子3(STAT3)表达对肿瘤浸润树突状细胞(TIDCs)成熟的影响。方法:在BALB/c小鼠大腿皮下接种小鼠恶性B细胞淋巴瘤A20细胞形成肿瘤,在肿瘤内注射STAT3基因特异的shRNA质粒并进行放疗;从肿瘤中富集TIDCs,采用荧光定量聚合酶链式反应(q PCR)方法检测TIDCs中STAT3 mRNA表达水平,蛋白染色方法检测磷酸化STAT3蛋白表达水平和DCs表面分子MHCII、CD40及CD80的表达,用流式细胞仪收集细胞染色数据。结果:放疗联合抑制STAT3表达显著降低肿瘤浸润TIDCs中STAT3 mRNA表达水平和STAT3蛋白磷酸化水平;与单独放疗相比,放疗联合抑制STAT3表达显著增加肿瘤浸润TIDCs表面分子MHCII、CD40、CD80及CD86的表达水平。结论:放疗联合抑制STAT3治疗表达能促进肿瘤浸润TIDCs的成熟。
Objective: To investigate the effect of combination of radiotherapy with STAT3 silencing on the maturation of tumor-infiltrating dendritic cells. Methods: Murine malignant B cell cell lymphoma A20 cells were subcetaneously implanted in BALB / c mice to grow tumors. Tumors were treated with a plasmid expressing STAT3-specific shRNA by intratumor injection and local radiation. Tumorinfiltrating dendritic cells( TIDCs) were enriched from tumors. STAT3 mRNA level in TIDCs was determined by q PRC,phospho-STAT3 level by intracellular staining and subsequent data collected by flow cytometry. The expression of MHCII,CD40,CD80 and CD86 were determined by extracellular staining and subsequent data collected by flow cytometry. Results: Combination of radiotherapy with STAT3 silencing inhibited STAT3 mRNA and decreased phosphorylation level of STAT3. Compared to radiotherapy alone,STAT3 silencing plus radiotherapy significantly upregulated the expression of MHCII,CD40 and CD80. Conclusions: Radiotherapy combined STAT3 inhibition promotes the maturation of tumor-infiltrating dendritic cells.

关键词(KeyWords)： 信号转导与转录激活因子3;放射疗法;淋巴瘤,B细胞;树突状细胞;肿瘤浸润
signal transducer and activator of transcription;radiotherapy;lymphoma,B cell;dendritic cells;tumor-infiltration

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81560482);; 贵州省“2011协同创新中心”支助项目[黔教合协同创新中心(2014)06];; 贵州省科技厅重大专项[黔科合计Z字(2012)4010];; 贵州省科技厅科技计划项目[黔科合LG字(2012)009]

作者(Author): 张启芳;吴昌学;禹文峰;官志忠;柏华;
ZHANG Qifang;WU Changxue;YU Wenfeng;GUAN Zhizhong;BAI Hua;Key Laboratory of Molecular Biology,Guizhou Medical University;Medical Experimental Center,Third Affiliated Hospital of Guizhou Medical University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2015.11.007

参考文献(References)：

• [1]Williams ME,Dreyling MH,Kahl BS,et al.Mantle cell lymphoma:report of the 2009 mantle cell lymphoma consortium workshop[J].Leuk Lymphoma,2010(3):390-398.
• [2]Martin PA,Chadburn,Christos P,et al.Intensive treatment strategies may not provide superior outcomes in mantle cell lymphoma:overall survival exceeding 7 years with standard therapies[J].Ann Oncol,2008(7):1327-1330.
• [3]Dreyling M,Amador V,Callana M,et al.Update on the molecular pathogenesis and targeted approaches of mantle cell lymphoma(MCL)-summary of the 12 annual conference of the EUROPEAN MCL NETWORK[J].Leuk Lymphoma,2014(4):1-26.
• [4]Chen Y,Wang M,Romaguera J.Current regimens and novel agents for mantle cell lymphoma[J].Br J Haematol,2014(1):3-18.
• [5]Foran JM,Cunningham D,Coiffier B,et al.Treatment of mantle-cell lymphoma with rituximab(chimeric monoclonal anti-CD20 antibody):analysis of factors associated with response[J].Ann Oncol,2000(Suppl 1):117-121.
• [6]Sabado RL,Bhardwaj N.Cancer immunotherapy:dendritic-cell vaccines on the move[J].Nature,2015(7543):300-301.
• [7]Palucka AK,Ueno H,Fay JW,et al.Taming cancer by inducing immunity via dendritic cells[J].Immunol Rev,2007(220):129-150.
• [8]Nagaraj S,Ziske C,Schmidt-Wolf IG.Dendritic cell,the immunotherapeutic cell for cancer[J].Ind J Med Resh,2004(4):133-138.
• [9]Palucka K,Hideki U,Joseph F,et al.Dendritic cells and immunity against cancer[J].Intern Med,2011(1):64-73.
• [10]Yu H,Kortylewski M,Pardoll D.Crosstalk between cancer and immune cells:role of STAT3 in the tumour microenvironment[J].Nat Rev Immunol,2007(1):41-51.
• [11]Steinbrink K,Wolfl M,Jonuleit H,et al.Induction of tolerance by IL-10-treated dendritic cells[J].J Immunol,1997(10):4772-4780.
• [12]Wang T,Niu G,Kortylewski M,et al.Regulation of the innate and adaptive immune responses by Stat-3 signaling in tumor cells[J].Nat Med,2004(1):48-54.
• [13]Williams L,Bradley L,Smith A,et al.Signal transducer and activator of transcription 3 is the dominant mediator of the anti-inflammatory effects of IL-10 in human macrophages[J].J Immunol,2004(1):567-576.
• [14]Gabrilovich DI,Chen HL,Girgis KR,et al.Production of vascular endothelial growth factor by human tumors inhibits the functional maturation of dendritic cells[J].Nat Med,1996(1):1096-1103.
• [15]Nefedova Y,Huang M,Kusmartsev S,et al.Hyperactivation of STAT3 is involved in abnormal differentiation of dendritic cells in cancer[J].J Immunol,2004(1):464-474.
• [16]Kitamura H,Kamon H,Sawa S,et al.IL-6-STAT3 controls intracellular MHC class II alphabeta dimer level through cathepsin S activity in dendritic cells[J].Immunity,2005(5):491-502.
• [17]Wang P,Xue Y,Han Y,et al.The STAT3-binding long noncoding RNA lnc-DC controls human dendritic cell differentiation[J].Science,2014(6181):310-313.
• [18]Galluzzi L,Kepp O,Kroemer G.Immunogenic cell death in radiation therapy[J].Onco Immunology,2013(10):26536.
• [19]Kroemer G,Galluzzi L,Kepp O,et al.Immunogenic cell death in cancer therapy[J].Annu Rev Immunol,2013(10):51-72.
• [20]Gao C,Kozlowska A,Nechaev S,et al.TLR9 signaling in the tumor microenvironment initiates cancer recurrence after radiotherapy[J].Cancer Res,2012(17):4440-4448.

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