nm23基因调控”EMT-CTC-OSM”级联抑制肺癌器官特异性转移的分子机制

课题组前期构建了2株不同转移潜能和4株不同器官特异性转移肺癌细胞株，证明nm23-H1抑制肺癌EMT，减少CTCs。本项目应用上述6株肺癌细胞株建立肺癌器官特异性转移瘤动物模型；应用ClearCell○RCX 循环肿瘤细胞分选回收系统，分选回收上述肺癌细胞株荷瘤鼠血液CTCs；应用miRNA和表达谱芯片筛选鉴定上述肺癌细胞株、CTC、移植瘤及器官特异性转移瘤组织中差异miRNA及差异基因并应用RT-PCR及Western blot验证；应用RT-PCR，Western blot检测上述肺癌细胞株、CTCs、移植瘤及器官特异性转移瘤组织中EMT相关基因miRNA及蛋白表达；应用生物信息学分析nm23-H1基因调控“EMT-CTC-OSM”抑制肺癌器官特异性转移的相关性。本项目对于阐明nm23-H1基因调控肺癌器官特异性转移的分子机制，发现逆转肺癌器官特异性转移的分子靶点提供理论基础及实验依据

This project plans to utilize the 6 lung cancer cell lines, which we previously established, to establish nude mice models with organ-specific metastasis； to use ClearCell® CX system to search and identify CTCs from the nude mice planted with the parent and organ-specific metastasis lung cancer cell lines； to use miRNA and gene array to detect the differential miRNAs and genes that relate to organ-specific metastasis of lung cancer from the lung cancer cell lines, CTCs, planted tumor tissues and the organ-specific metastasis tumor tissues from the nude mice bearing the lung cancer cells. We will further detect the key genes and proteins expression by RT-PCR, Western blot，which relate to EMT and organ-specific metastasis in the lung cancer cell lines, CTCs, planted tumor tissues and the organ-specific metastasis tissues from the nude mice bearing the lung cancer cells. Finally, we will use bioinformatics to analyze the relationship how the nm23-H1 gene regulate “EMT-CTC- OSM ”cascade, and reverse organ-specific metastasis of lung cancer, and verify the key roles of nm23-H1 gene suppress lung cancer organ-specific metastasis. Overall, findings from the project will not only provide the novel molecular mechanisms of organ-specific metastasis, but also identify novel targets in clinical in the future.