循环肝癌细胞关键耐药靶点的筛选及其干预

我们前期研究发现，肝癌患者外周血中的循环肝癌细胞（Circulating Liver Cancer Cells，CLCC）具有异质性，与肝癌耐药及转移复发密切相关；筛选CLCC耐药亚群，找到特异性耐药靶点是克服肝癌耐药的关键。阿司匹林可显著减少部分肝癌患者CLCC，并与索拉菲尼及化疗药（5-FU/ADM）具有协同或增敏作用，其机制不明。本课题将在大量前期工作基础上，1. 利用我们自主研发的免疫磁珠微流控CLCC分选系统，精确筛选化疗及索拉菲尼耐药肝癌病人CLCC，利用单细胞转录组扩增测序技术，探索其共性耐药分子标签，并通过基因转染、RNAi、基因敲除动物、组织芯片等方法明确验证。2. 筛选阿司匹林干预后耐药逆转CLCC，检测耐药相关靶点及信号通路变化并验证。通过本研究得到的特异性肝癌耐药分子可用于临床化疗及索拉菲尼敏感人群的筛选，并为今后阿司匹林逆转肝癌耐药的大规模临床试验奠定理论基础。

In our previous studies, we found circulating liver cancer cells (CLCC) resisted to chemotherapy or Sorafenib, and may lead to the failure of treatments and poor prognosis. The heterogeneity of CLCC has been confirmed, and screening the subpopulations of CLCC and its key genes of chemo-resistance could be very helpful to the treatments of liver cancer patients. Aspirin can decrease CLCC ratio in some patients and increase the effects of sorafenib. In this study, we plan to 1, Selecting CLCC from patients under different treatments like sorafenib or combined chemotherapy (with/or without Aspirin) using our solid screen system based on immunomagnetic beads microfluidic chip. Exploring the key gene expression profiling of drug-resistance using Single-Cell genomics system, and then confirming the results in vitro and in vivo. 2) Reveal the underlying mechanism in anti-drug resistance of Aspirin. In summary, we try to create a novel stratagem to predict liver cancer patients sensitive to chemo-therapy or sorafenib, and to help some of those non-sensitive patients.