【作者】 陈静；

【导师】 俞磊；

【作者基本信息】 华东师范大学 ， 生物化学与分子生物学， 2014， 硕士

【摘要】 肿瘤微环境指的是肿瘤局部浸润的间质细胞,免疫细胞和它们所分泌的活性介质与肿瘤细胞共同组成的局部的内环境。肿瘤特殊的微环境导致肿瘤有其独特的特征,如抵抗细胞死亡,细胞能力异常,基因组不稳定和持续增殖的信号,回避生长抑制,免疫逃逸,无限的复制能力,特异的血管生成,浸润和转移等。这些特征导致肿瘤的传统治疗方法如手术切除,放疗和化疗效果不理想,病人预后较差,易复发和转移。肿瘤细胞能促进血管的新生,有着入侵和转移的能力,这些肿瘤行为受到肿瘤微环境中的非肿瘤细胞的调节。肿瘤内部混乱的血管构造导致很多区域的缺氧坏死,血管对肿瘤药物的吸收和运输存在重要影响,所以目前已有针对肿瘤的微环境如抑制血管新生等治疗手段。而成纤维细胞是肿瘤间质细胞中的一大类,对肿瘤细胞起着至关重要的支撑作用。相对于复杂的肿瘤细胞,有着稳定遗传物质的成纤维细胞显然更容易成为研究肿瘤药物的靶点。所以本研究选取三种不同的肿瘤模型,研究其血管密度和胶原纤维含量的差异。肿瘤在生长过程中,不同的体积也将对药物的运输产生影响。肿瘤体积过大,则往往内部更多的处在缺氧缺血的状态,而这种缺血缺氧导致的酸性,炎症环境,都将会对药物的运输有着重要的影响。人体内肿瘤的倍增时间往往与病人的生存时间,手术预后等有着明显的关系,裸鼠皮下移植瘤的生长速度也各有不同。这些不同的生长速度同样影响着肿瘤的内部形态和组成,所以本研究也将评估三种肿瘤模型中肿瘤的不同体积和生长速度。本研究采用的方法为在裸鼠皮下注射成系肿瘤细胞悬液的方法来构建肿瘤模型。分别用HT29细胞、U87-MG细胞、A549细胞皮下移植瘤模拟结肠癌、脑胶质瘤、非小细胞肺癌这三种肿瘤的微环境。通过将CD31免疫组化染色、胶原纤维MASSON三色染色方法直观的观察肿瘤内部的血管和胶原纤维含量,再通过MVD法和图像处理软件将图片半定量,从而得到较为精确的数据。再将数据进行统计学分析,从而得出结论：这三种肿瘤模型在生长速度、微血管密度、胶原纤维含量上面都有显著差异。为使用这三种肿瘤模型的药物筛选实验提供指导。更多还原

【Abstract】 Tumor microenvironment refers to regional invasion interstitial cells of tumor, immunocytes and the local inner environment made by their secretion of active medium and the tumor cells.Special tumor microenvironment of tumor lead to unique characteristics, such as resistance to the death of cell, abnormal cell capacity, the signal is not stable and continuous appreciation of the genome, avoid growth inhibition, immune escape, unlimited replication ability, specific angiogenesis, invasion and metastasis, and so on.These characteristics, leading to the traditional treatment of tumors such as surgery, radiotherapy and chemotherapy effect is not ideal, patient prognosis is poorer, easy recurrence and metastasis.Tumor cells can promote the blood vessels of the newborn, has the ability of invasion and metastasis, these behaviors of tumor always adjust by cells in the tumor microenvironment without tumor cells.Inside tumor vascular structure led to many areas of necrosis by the lack of oxygen, blood vessels has an important influence in absorption and transportion of tumor drug, so there are some treatments to against tumor microenvironment such as inhibiting angiogenesis.The fibroblast is one of the interstitial cells of tumor, it plays an important supporting roles of the tumor cells.Relative to the complex of tumor cells, it has a stable genetic material into a fibrous cells obviously, it’s clearly that it is easy to be the target of research on the tumor drugs.So the experiment choose three kinds of different tumor model, have a study on the density of blood vessels and collagen fiber content differences. During the growth of tumor, different volume will also impact on the transport of drugs.When the volume is too large, it means the internal of tumor is more often in a state of hypoxia ischemia, and the acidic and inflammation of the environment caused by it always has an important influence on the transport of drugs.The doubling time of tumors in the human body always has obvious relationship with the survival time of patients and surgery prognosis,the subcutaneous transplantation tumor growth speed of nude mice is vary too.These different growth speed also affect internal morphology and composition of the tumor, so this study will also study the different size and growth speed of the three tumor model.This study adopts the methods in nude mice subcutaneously into cell suspension method to build the model of the tumor.Use the subcutaneous transplantation tumor of HT29, U87-MG and A549to simulated colon cancer, glioma, non-small cell lung cancer of the three tumor microenvironment.Use the methods of CD31immunohistochemical staining and MASSON three color dyeing to intuitive look inside the tumor blood vessels and collagen fiber content, then the MVD method and image processing software will be semi-quantitative images, to get more accurate data. Again the data will be used statistical analysis, finally draw the conclusion:the three tumor model have significant differences all in the growth speed, the microvascular density and the collagen fiber content, The results of this study will guide the others who use these three kinds of tumor drug screening experiment.更多还原