基于小鼠肺脱细胞基质人乳腺癌肺转移体外三维培养模型的建立

Breast cancer is a serious threat to women's life and health, and lung metastasis is the leading cause of breast cancer-related death. Lack of an ideal model for mimicking the breast cancer lung metastasis is still a bottleneck restraint for the study of its mechanism. We plan to use the mouse lung acellular matrix to mimic the pre-metastatic lung microenvironment of human breast cancer in vitro, and then engraft human breast cancer cell line originated from the MDA-MB-231 with high metastasis potential onto the mouse matrix through trachea or pulmonary artery inoculation followed by culture in perfusion bioreactor to establish a novel human tumor metastasis model. At the same time, we establish the classic human breast cancer metastasis model in SCID mouse. And then we compare the cell proliferation kinetics, tumor histomorphology, ultra-microscopic structure of tumor, invasive ability of tumor cells and the metastasis associated proteins profiling between the two models. Furthermore we are going to compare the gene-expression profiling of migratory tumor cells and drug susceptibility of lung metastasis in the two models in order to evaluate the practical application value of this new mouse lung acellular matrix based tumor metastasis model. This work will provide a new tool for studying breast cancer in an ex vivo three-dimensional environment that closely simulates the actual tumor metastatic microenvironment, this model can be used to improve our understanding of the tumor microenvironment and mechanism in breast tumor development.

乳腺癌严重着威胁女性生命健康，肺转移是患者死亡的最主要原因。目前尚缺乏用于乳腺癌肺转移研究的理想实验模型。我们拟利用小鼠肺脱细胞基质在体外模拟人乳腺癌肺转移前微环境，经气管和经肺动脉注射两种途径，把高转移潜能的人乳腺癌MDA-MB-231细胞接种在小鼠肺脱细胞基质中，并在带有氧合装置的连续灌注培养反应器内培养，建立人乳腺癌细胞株在小鼠肺脱细胞基质中成瘤模型，同时建立经典的SCID小鼠肺转移动物模型。并对两个模型中转移瘤细胞增殖动力学、组织形态学、超微结构、细胞侵袭性以及与肿瘤转移相关蛋白的表达水平进行比较。以基因表达谱芯片检测肿瘤细胞在两个模型中基因表达情况，通过经典抗乳腺癌药物敏感实验，观察各化疗方案对肿瘤细胞在两个模型中转移瘤成瘤能力的影响，评估基于小鼠肺脱细胞基质人乳腺癌肺转移模型在乳腺癌肺转移体外研究中的应用价值，为进一步探讨乳腺癌肺转移的生物学机制提供理想的体外三维培养模型。

乳腺癌严重威胁着女性健康，肺转移是其主要致死因素，目前尚缺乏乳腺癌肺转移理想的体外研究模型。为探索人乳腺癌细胞在脱细胞化的鼠肺中成瘤的可行性，一定程度上模拟乳腺癌肺转移，我们取成年SD大鼠心肺组织，对离体的鼠肺行灌注法脱细胞处理，建立、完善了大鼠肺灌注脱细胞装置及基本操作流程，并对大鼠肺脱细胞基质制备参数进行了优化。进而将人乳腺癌细胞株Mcf-7经气管种植于脱细胞化鼠肺基质中，并将其置于特制的生物反应器中灌注培养1-2周，在体外培养出了真正的三维人乳腺癌组织。我们进一步经肺动脉注射途径将人乳腺癌细胞Mcf-7接种在脱细胞化的鼠肺中，期望模拟人乳腺癌血行转移在肺内成瘤过程，目前尚未获得满意结果。在该研究中我们还将人肺癌细胞H460经气管接种在脱细胞鼠肺基质中，也成功在体外培养出了三维人肺癌组织。这一研究结果说明鼠肺脱细胞基质可以作为一种较为理想的体外肿瘤组织三维培养基质支架。目前已有2篇中文文章及1篇英文文章在投。