sCLU通过CDC25A诱导胆管癌化疗耐药机制及其作为联合治疗靶点的潜在价值研究

吉西他滨是胆管癌化疗的首选药物，但临床存在广泛耐药且机制不明，推测可能与肿瘤细胞周期阻滞的修复相关。我们前期基于CRISPR/Cas9的sgRNA文库筛选发现，sCLU为吉西他滨联合抑制胆管癌细胞增殖的潜在靶点，sCLU与细胞周期关键调控“开关”蛋白CDC25A存在互作，结合多组学分析提示sCLU可能通过稳定CDC25A，阻碍吉西他滨对细胞周期的抑制，从而诱导化疗耐药。为验证上述假设，本项目拟通过细胞实验验证sCLU与CDC25A的互作关系及对胆管癌细胞恶性表型和吉西他滨耐药的影响，从分子水平揭示sCLU通过CDC25A促进周期转化进程诱导吉西他滨耐药的机制，采用PDX动物模型探索sCLU特异性抑制剂OGX-011联合治疗胆管癌的有效性，明确sCLU作为联合治疗靶点的潜在价值，阐明胆管癌化疗耐药机制，为逆转胆管癌吉西他滨化疗耐药提供新的思路。

Although gemcitabine is the preferred agent of cholangiocarcinoma chemotherapy, the mechanism underlying extensive drug resistance is unknown. It is reported that repairing of the tumor cell cycle arrest may play a role. Our previous CRISPR / Cas9 sgRNA library screening revealed sCLU is a potential marker that can synergistically facilitate the anti-proliferation effect of gemcitabine, by interacting with CDC25A, which is a key cell cycle regulator "switch" protein. Results from multi-omics analysis suggest that through stabilizing CDC25A, sCLU diminishes the cell cycle-inhibiting effect of gemcitabine, thus inducing drug resistance. In order to verify this hypothesis, in vitro study will be applied to demonstrate the interaction pattern of sCLU and CDC25A: by interacting with CDC25A, sCLU can promote the cell cycle progress which can induce gemcitabine resistance. Corresponding cancer cell malignant phenotype change, as well as gemcitabine resistance, will be studied. Lastly, the PDX animal model will be used to study the effectiveness of sCLU-specific inhibitor OGX-011, to further clarify whether sCLU is a promising target, hence providing new therapeutic strategy for gemcitabine-resistant cholangiocarcinoma.