基于脱细胞支架的肝癌三维动态培养模型及个体化药物敏感谱的建立

Interaction between the hepatoma cells and microenvironment plays an important role in malignant phenotype of the liver cancer. However, the existing in vitro liver cancer models are unable to simulate the 3D microenvironment in vivo, resulting in poor intracorporal replication of research findings especially the sensitivity of anti-cancer drugs, and significantly reducing the research efficiency. Therefore, to establish a more emulational 3D liver cancer model in vitro is very important for basic and clinical research of liver cancer. Our previous research and other literature has demonstrated that decellularized liver scaffolds, which provide natural extracellular matrix component, three-dimensional porous structure and microvascular network for liver tissue reconstruction in vitro, could be the ideal material for liver tissue engineering. This project intends to establish a 3D dynamic liver cancer model on the basis of decellularized liver scaffold. This model will potentially provide a more emulational research platform for development, invasion, metastasis, and drug resistance of liver cancer. This model will then be used for cultivation of patients’ primary liver cancer cells, and establishing an individualized drug sensitive spectrum through anti-cancer drug sensitive test. This project chould provide experiment basis for the individualized treatment of liver cancer.

肿瘤微环境对肝癌的恶性表型具有重要作用。现有的体外肝癌模型对体内三维微环境的仿真性不足，导致研究结果的体内复制性差，极大降低了研究效率。因此，建立一套仿真度更高的体外肝癌模型，对于优化肝癌基础及临床研究具有重要意义。项目组前期研究及文献均证实，肝脏脱细胞支架具有天然的细胞外基质成分、三维多孔结构和微血管网络，生物相容性良好，是体外重建肝脏组织的理想材料。我们设想利用肝脏脱细胞支架能够构建高度仿真的肝癌三维动态培养模型，并以此为平台建立个体化的抗癌药物敏感谱。为此，本项目拟：（1）研究确定关键技术参数，构建基于肝脏脱细胞支架的肝癌三维动态培养模型；（2）通过对肝癌发生发展、侵袭转移、耐药性等生物学行为进行研究，评价模型的可行性和优势；（3）利用成功构建的肝癌三维动态模型培养患者原代肝癌细胞，结合药敏试验建立个体化的抗癌药物敏感谱，为实现肝癌的个体化治疗奠定实验基础。

肿瘤微环境对肝癌的恶性表型具有重要作用。现有的体外肝癌模型对体内三维微环境的仿真性不足，导致研究结果的体内复制性差，极大降低了研究效率。因此，建立一套仿真度更高的体外肝癌模型，对于优化肝癌基础及临床研究具有重要意义。本项目组首次提出利用肝脏脱细胞支架构建高度仿真的肝癌三维动态培养模型。研究证实：1）成功制备了大鼠肝脏、脾脏及人肝脏脱细胞支架，验证了脱细胞支架用于细胞体外三维培养的可行性。2）完成肝癌细胞的体外三维动态培养模型的构建，为肝癌生物学行为研究提供仿真度更高的体外研究平台。3）证实非酒精性脂肪肝细胞外基质Collagen I 调控Integrin β1/FAK 信号通路促进肝癌细胞增殖。4）采用光化学交联法优化肝脏脱细胞支架的生物力学性能，提高支架的抗降解能力，增加脱细胞支架内血管生成。本项目的成功实施，为肝癌发生发展、侵袭转移、耐药性等生物学行为研究，以及抗癌药物筛选提供仿真度更高的体外研究平台；也为组织工程肝脏的体外构建和体内植入奠定了实验基础。