特异性抑制肺癌干细胞化合物的作用机制研究

Even after modern cancer treatments, lung cancer can recur as a result of the presence of lung cancer stem cells. Although the scientists has discovered lung cancer stem cell surface markers and differentiation, proliferation regulatory factors, such as CD133, c-kit, the Wnt, Notch, Oct4 which are the same with normal stem cells, we have not been unable to design drugs to target these markers. So,we need to screen compound libraries with lung cancer stem cell-specific target compounds which can then be used as clues to eluciate lung cancer stem cell specific differentiation and proliferation regulation mechanisms. We have successfully generate lung cancer stem cells by using will induced pluripotent stem cell (iPS) technology. Taking advantage of this cell model, we got compounds which can specificlly inhibit lung cancer stem cells. Based on the findings, we plan to use two-dimensional protein electrophoresis combined with protein mass,compounds labeled with biotin and co-precipitation technology to find out the targets of the compounds. And then by using downstream gene expression analysis and RNA interference technology to clarify its mechanism and revealing the new regulation mechanis of lung cancer stem cell differentiation and proliferation. Finally we will test the compounds' anti-tumour ability by using animal model and patients' primary lung cancer stem cells. This study is expected to provide new lung cancer stem cell specific targets and the new mechanisms of lung cancer stem cell differentiation and proliferation.

肺癌干细胞的残存是肺癌复发的根源。迄今发现的肺癌干细胞表面标记和增殖调控因子如，CD133,c-kit,Wnt,Notch,Oct4都与正常干细胞相同，无法以这些已知靶点设计药物。因此需要从化合物库中筛选具有肺癌干细胞特异性靶点的化合物，并以此为线索发现新的肺癌干细胞特殊的分化，增殖调控机制。申请者利用诱导性干细胞（iPS）技术建立了肺癌干细胞系，通过高通量筛选,得到了不抑制肺癌细胞和正常胚胎干细胞而特异性抑制肺癌干细胞的化合物。本研究以此为基础，拟通过二维蛋白电泳结合蛋白质谱，化合物同位素标记和共沉淀等技术，明确所得化合物的作用靶点。再通过对下游基因的表达分析，干扰RNA等技术阐明其作用机制,最后以病人原代肺癌干细胞和小鼠肺癌转移模型证明所得化合物的抗肿瘤功能。本研究有望发现新的肺癌干细胞的作用靶点，并揭示调控肺癌干细胞分化、增殖的新机制,为基于肺癌干细胞的药物治疗提供理论依据。

肺癌的复发是肺癌治疗的瓶颈，在这个过程中，肺癌干细胞起了很重要的作用。本研究首先利用诱导性干细胞（iPS）技术将肺癌肿瘤细胞诱导成肺癌肿瘤干细胞的方法，其次利用诱导性肺癌干细胞样细胞hLCSC-A549和hLCSC-CH27成功构建了GFP稳定表达的诱导性hLCSC细胞株，并进一步利用诱导的肺癌干细胞筛选得到6种有效化合物，研究了化合物LCSC-K1调节hLCSC分化和增殖的机制，着重研究细胞自噬诱导剂LCSC-K1对细胞自嗜关键信号分子mTOR和MAPK介导的信号途径的影响，并阐明Atg5非依赖性自嗜调控线粒体清除和细胞重编程重要机制，为基于肺癌干细胞的药物治疗提供理论依据。. 在寻找新抗肿瘤靶点的研究过程中，肿瘤干细胞的发现给肿瘤治疗带来了新的曙光，但关于肿瘤干细胞的来源及调控机制还不清楚。研究证明了CREPT在干细胞中的作用以及对肿瘤干细胞的影响，并研究了CREPT在肺癌发生中的作用机制，为临床治疗提供更多思路。