抗非小细胞肺癌细胞及其干细胞的功能性脂质体构建及治疗机制研究

Scientific problems: Lung cancer is one of the world's highest morbidity and mortality of malignant tumor. Patients with lung cancer in 80%-85% is non-small cell lung cancer, and lost the chance of operation due to the initial diagnosis in patients with most of the time have been in the last stage. However, the effect of radiotherapy and chemotherapy to treat lung cancer is not ideal. Radiotherapy is difficult to eradicate the invasion and metastasis of lung cancer cells. The chemotherapy without selectivity kill cancer cells and cancer stem cells at the same time also caused damage to normal cells. The results of literature investigations indicated that how to improve the anticancer drug targeting to lung tumor tissue and lung tumor stem cell still remained to be the major clinical difficult issues, and therefore, to be the unsolved scientific questions. Objectives: A novel kind of targeting anticancer liposomes is prepared by conjugating a ligand onto the liposome surface for enhancing permeability of drug to lung cancer tissue, and enhance anticancer drug targeting to lung caner cell and increase the drug concentration onto lung cancer cells and lung cancer stem cells. Research contents: (1) To develop a novel kind of targeting liposomes and characterization; (2) To culture lung tumor cells and lung tumor stem cells; (3) To assay recognition, appraisement and charactrisition on lung tumor stem cells; (4) Anti-proliferation and targeting function on the lung cancer cells and lung cancer stem cells; (6) Research on molecular regulation mechanism; (5) To evaluate anti-tumor effects in vivo on bearing A549 lung cells animal model. Significance: It is theory and clinical significance to explore the therapy strategies on curing refractoriness tumor and relapse of treated lung cancers.

科学问题：肺癌发病率和死亡率高，病人生存期短，且多数患者初次诊断时已处于晚期，失去手术机会，然而放疗化疗难以根除浸润转移的肺癌细胞，在杀伤癌细胞的同时不能根除肺肿瘤干细胞。因此，如何使化疗药物在到达肺组织后，靶向浓集于肺肿瘤并能诱导肺肿瘤细胞及其干细胞凋亡，同时降低副作用是仍待解决的重要科学问题。目的：以新疆天山雪莲中抗肿瘤有效成分粗毛豚草素为药物，以叶酸为肺癌靶向功能配体修饰到脂质体表面上，表没食子儿茶素没食子酸酯为肺肿瘤干细胞诱导凋亡剂包载在脂质体内，构建的全新的具有肺癌靶向功能的载药脂质体，可靶向浓集于肺肿瘤诱导肺肿瘤细胞及其干细胞的凋亡，同时降低副作用。内容：通过（1）脂质体构建与表征；（2）肺肿瘤细胞及其干细胞的培养；（3）肺肿瘤干细胞鉴定与表征；（4）对肺肿瘤细胞及其干细胞毒性与靶向性研究 ；（5）治疗作用机制研究；（6）在动物体内疗效评价。意义：具有重要的理论意义。

项目背景：肺癌发病率和死亡率高，病人生存期短，且多数患者初次诊断时已处于晚期，失去手术机会，然而放疗化疗难以根除浸润转移的肺癌细胞，在杀伤癌细胞的同时不能根除肺肿瘤干细胞。因此，如何使化疗药物在到达肺组织后，靶向浓集于肺肿瘤并能诱导肺肿瘤细胞及其干细胞凋亡，同时降低副作用是仍待解决的重要科学问题。.主要研究内容：（1）天山雪莲抗癌成份筛选及对肺癌细胞EMT调控作用机制的研究；（2）粗毛豚草素肺靶向脂质体的构建及药效学评价研究；（3）天山雪莲功能化纳米载体构建及其对肺癌EMT的抑制作用研究；（4）抗肺肿瘤靶向脂质体的构建及其效应评价研究。.重要结果及关键数据：（1）运用系统药理学方法筛选了天山雪莲活性活性化合物并对其机制进行剖析，实验证明4种成分均能抑制A549细胞、C6细胞及PC-3细胞增殖，其机制可能是抑制了PI3K-Akt通路，抑制Akt的磷酸化。（2）发现天山雪莲粗毛豚草素对A549细胞上皮间质转化具有抑制作用，在此基础上，以叶酸为肺癌靶向功能配体修饰到载体表面上，表没食子儿茶素没食子酸酯为肺肿瘤干细胞诱导凋亡剂包载在脂质体内，构建具有肺癌靶向功能的全新载药脂质体。所得制剂可靶向浓集于肺肿瘤诱导肺肿瘤细胞及其干细胞的凋亡，同时降低副作用。实验中建立粗毛豚草素及EGCG体外含量测定方法学稳定可靠。（3）构建天山雪莲功能化纳米载体，发挥出显著的抗增殖效果和促凋亡作用，并能够显著抑制细胞内活性氧的表达和A549细胞的侵袭转移，阐明天山雪莲功能化纳米载体通过调节N-cadherin、Vimentin与E-cadherin蛋白的表达来发挥抑制A549肺癌细胞上皮间质转化的作用。（4）制备得到紫花牡荆素靶向脂质体，显示出更有效渗透进肿瘤球内部的能力，进而提升抑制A549细胞增殖效果。.科学意义：本项目所构建的靶向递药载体以及抗肺肿瘤的EMT作用机制研究结果，为临床治疗肺肿瘤提供更多的实验数据及研究基础，为进一步开发天山雪莲奠定良好基础和临床前景。