Angiostatin(K1-3)和endostatin裸DNA联合治疗人脑胶质瘤的实验研

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基本信息

  • 批准号:
    39970854
  • 项目类别:
    面上项目
  • 资助金额:
    11.0万
  • 负责人:
  • 依托单位:
  • 学科分类:
    H3505.抗肿瘤药物药理
  • 结题年份:
    2002
  • 批准年份:
    1999
  • 项目状态:
    已结题
  • 起止时间:
    2000-01-01 至2002-12-31

项目摘要

Aim In order to investigate the feasibility and characteristic of antiangiogenesis, the research of treatment of glioma were conducted. Method: Human angiostatin K(1-3) and endostatin gene fragments were cloned and sequenced by RT-PCR methods. Construction of angiostatin K(1-3) or endostatin eukaryotic expressed vector,which angiostatin K(1-3) and endostatin cDNA were linked with human Ig Kappa-chain secretive singal by PCR, the PCR product was cloned into eukaryotic expressed vector to construct pcDNA-SAK(1-3) or pcDNA-S- endostatin.The above eukaryotic expression vectors were transfected into human SHG44 glioma cell by lipofectamine method and the positive clones were selected by G418. The biological characters of glioma cells in experimental group, empty vector group and control group were examined by electron microscope, flow cytometry and cell growth rate counting, and there were no differences of cell cycles in these three groups. Immunohistochemistry, immunofluorescence,western blot and hybridization in situ confirmed that the protein of angiostatin K(1-3) and endostatin were expressed in the experimental group. The SHG44 cells were planted under the skin of nude mice to eastablish nude mice model and the tumorigenesis of the cells were observed and compared among these three groups.The nude DNA packaged by lipofactamine was injected to the tumor,the effects of angiostatin K(1-3) and endostatin protein to the nude mice were discussed. Results:Angiostatin K(1-3) and endostatin gene fragements were got and sequenced right. The pcDNA-SAK(1-3) or pcDNA-S-endostatin were constructed, which were correct by sequenceing and restriction enzyme analysis. The expressed angiostaitn K(1-3) or endostatin can inhibit chicken-embryo angiogenesis in vivo and didn't affect the biological activities of SHG44 gliomas cells. The tumorigenesis of experimental group cells compared with that of empty vector group or control group was greatly reduced. The tumor necrosis rate of experimental group was obviously higher than that of empty vector group or control group.The blood vessel numbers of experimental group were lower than those of empty vector group or control group. Conclusion:The above results illustrated that it is feasible to treat the human glioma under the skin of nude mice with the eukaryotic expressed angiostatin K(1-3) or endostatin nude cDNA. The tumor growth suppression in vivo was caused by angiostatin K(1-3) or endostatin due to its antiangiogenesis and tumor necrosis. Our experimental studies lay the preliminary foundation for further treatment of glioma and the other solid tumors by antiangiogenesis methods.
将构建在真核表达载体上的angiostatin(K1-3)和endostatin分别或联合注入荷脑胶柿雎闶蟾骨弧⒕猜龊土鎏迥冢τ妹庖咦榛⒃辉咏弧CR和Southern Blot检测其体内泶铮教致鉊NA注射后对体内器官的影响和抑瘤性,检测血液内各项生化指标的改变,为抗血管生成基因药物治疗人脑胶质瘤奠定理论和实践基础。这方面研究有重要临床应用价值。

结项摘要

项目成果

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其他文献

南昌市2014—2017年活禽市场禽流感病毒监测
  • DOI:
    --
  • 发表时间:
    2020
  • 期刊:
    中华实验和临床病毒学杂志
  • 影响因子:
    --
  • 作者:
    冯道斌;宋文涛;倪贤生;吴景文;王艳晴;陈胜恩;陈海婴;刘明斌
  • 通讯作者:
    刘明斌
南昌市活禽市场禽流感病毒及从业人员特征分析
  • DOI:
    --
  • 发表时间:
    2016
  • 期刊:
    中华疾病控制杂志
  • 影响因子:
    --
  • 作者:
    谢文静;胡茂红;宋文涛;吴景文;夏文;陈盛恩;陈海婴;刘明斌
  • 通讯作者:
    刘明斌

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课题项目:调控A型流感病毒诱导IFN-β表达的机制研究

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本研究聚焦于TRIM2蛋白在A型流感病毒诱导的IFN-β表达中的调控机制。A型流感病毒是全球性健康问题,其感染可导致严重的呼吸道疾病。IFN-β作为关键的抗病毒因子,其表达水平对抗病毒防御至关重要。然而,TRIM2如何调控IFN-β的表达尚未明确。本研究假设TRIM2通过与病毒RNA或宿主因子相互作用,影响IFN-β的产生。我们将采用分子生物学、细胞生物学和免疫学方法,探索TRIM2与A型流感病毒诱导IFN-β表达的关系。预期结果将揭示TRIM2在抗病毒免疫反应中的作用,为开发新的抗病毒策略提供理论基础。该研究对理解宿主抗病毒机制具有重要科学意义,并可能对临床治疗流感病毒感染提供新的视角。

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科学问题:TRIM2如何调控A型流感病毒诱导的IFN-β表达?
前期研究:已有研究表明TRIM2参与抗病毒反应,但其具体机制尚不明确。
研究创新点:本研究将深入探讨TRIM2在IFN-β表达中的直接作用机制。
技术路线:包括病毒学、分子生物学、细胞培养和免疫检测技术。
关键技术:TRIM2与病毒RNA的相互作用分析,IFN-β启动子活性检测。
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        graph TD
          A[研究起始] --> B[文献回顾与假设提出]
          B --> C[实验设计与方法学准备]
          C --> D[A型流感病毒感染模型建立]
          D --> E[TRIM2与病毒RNA相互作用分析]
          E --> F[TRIM2对IFN-β启动子活性的影响]
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          H --> I[数据收集与分析]
          I --> J[结果解释与科学验证]
          J --> K[研究结论与未来方向]
          K --> L[研究结束]
      
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