【作者】 王晶；

【导师】 李凯； 魏熙胤；

【作者基本信息】 天津医科大学 ， 肿瘤学， 2007， 硕士

【摘要】 背景：近年来，全世界肺癌发病率、死亡率持续上升，已经排在各种恶性肿瘤的第一位。非小细胞肺癌(Non-small cell lung cancer，NSCLC)占其中的80％左右，临床确诊时50％的患者已发生远处转移，且相当一部分是常规检查难以发现的，造成术后很快复发。目前，治疗远方转移的主要方法为化疗，但治疗效果差，容易发生耐药。因而靶向治疗成为当今研究的热点，这之中以内皮抑素较为理想，许多国内、外学者尝试将其用于临床治疗。但对其作用机理及疗效的预测都还知之甚少。目的：观察经不同剂量Endostatin干预后，移植瘤的生长及与肿瘤增殖、转移、凋亡及新生血管形成等方面相关的生物标志物的表达和变化，初步探讨Endostatin的体内作用机制、并确立肿瘤组织与外周血的联合检测标记物群，期望在临床上更易实施和动态观察。方法：给予荷瘤鼠不同剂量Endostatin干预后进行以下检测：1．应用免疫组织化学方法检测肿瘤组织中Survivin、VEGF、COX-2、C-erbB2、CD105蛋白表达及MVD数量。2．应用流式细胞术检测外周血内皮细胞相关标记(CD146、CD105单标记及双标记的表达)；Real-time PCR法检测CD146、CD105mRNA的含量。结果：Endostatin干预后，Survivin、VEGF、COX-2、C-erbB2、CD105蛋白表达及MVD数量一致性下降，并在一定范围内具剂量依赖性；外周血中内皮细胞数量随之下降，其程度在一定范围内也具剂量依赖性，其相应的mRNA呈现一致变化，并具正相关性；免疫组化指标与外周血指标间呈现明显正相关性。结论：1．Endostatin可抑制肿瘤生长并可降低移植瘤中的VEGF、Survivin、C-erbB2、COX-2、CD105的表达和MVD的数量，说明Endostatin可通过作用信号传到通路中多个环节，达到抑制新生血管形成的最终效果。2．经Endostatin干预后动物外周血中内皮细胞数量随之下降，下降程度在一定范围内呈剂量依赖性，其mRNA呈一致性变化。这可能与Endostatin抑制血管内皮细胞活化并诱导其凋亡有关。3．循环血管内皮细胞与其相应的瘤组织中Survivin等指标间呈正相关性，提示其可能替代肿瘤组织中的部分指标检测，作为预测疗效果和预后指标应用于临床。更多还原

【Abstract】 Background:Recently, the incidence and mortality of lung cancer in whole worldcontinue to increase and be highest in all of malignancy. About 80% ofthem are no small cell lung cancer. And 50% of patients given the diagnosisof NSCLC belong to advanced disease. They usually have several metastasiswithout discovered by routine medical examination. Thus, the NSCLCrelapse quickly after operation. Now, chemotherapy plays a key role intreatment of advanced NSCLC. But, efficacy of chemotherapy is verylimited. The phenomenon of drug resistant in NSCLC is very common. So,the target drugs become the hotpoint of research domestically andinternationally. Endostatin is the most promising drug among them, manydoctors try to use it in clinic. But we know a litter about its michenismand estimating efficacy.Objective:Observing the growth of xenograft tumor of Calu-6 in mice and the changeof biological marker which reflect proliferation、metastasis、apoptosis、angiogenesis of tumor treated by different dose Endostatin. Investigatethe mechanism of Endostatin and establish associated testing with tissueand peripheral blood in order to put it in clinical work and kinesisobservation.Methods: Treated with different dose Endostatin on xenograft tumor of Calu-6in mice, then detect markers like followings:1. To examine tumor tissue. Such as Survivin、VEGF、COX-2、C-erbB2 byimmunohistochemistry.2. circulating endothelial cells in peripheral blood by flow cytometry;its content of RNA by RT-PCR、real-time PCR.Recult:After teated with Endostatin, the express of Survivin protein、C-erbB2protein、COX2 protein、VEGF protein and MVD descend consistent, and thisdegression is dose dependent and the CECs in peripheral blood are diminish,this decreasion is dose dependent. Positive correlation are foundbetweenits circulating endothelial cells and its mRNA, circulatingendothelial cells and immunohistochemistry marker.Conclusions:1. Endostatin can inhibit tumors’ growth and diminish expression ofSurvivin protein、C-erbB2 protein、COX2 protein、VEGF protein andMVD.2. The CECs in peripheral blood are diminish, this decreasion is dosedependent. Positive correlation are found betweenits circulatingendothelial cells and its mRNA.更多还原