shRNA干扰新型肿瘤抗原Eps8对白血病TKIs耐药的逆转作用及其机制研究

白血病患者对酪氨酸激酶抑制剂（TKIs）产生耐药使其临床应用价值受到限制。本课题组收集白血病患者骨髓标本进行研究，发现表皮生长因子受体通路底物8（Eps8）的表达水平与化疗药物的疗效呈负性相关，提示Eps8有可能参与白血病耐药性的发生。Eps8作为一种新型肿瘤抗原参与多种实体瘤的恶变过程，为预后不良的指标之一。我们在前期工作中成功构建了含有异常Abl基因的前B淋巴细胞白血病细胞株，该细胞株对TKIs STI571耐药，并且Eps8基因高表达，是研究白血病对TKIs耐药的良好模型。为了验证Eps8与白血病TKIs耐药之间的关系我们以上述白血病耐药细胞为靶细胞，应用shRNA靶向沉默Eps8基因表达，观察其逆转TKIs耐药的效应，并进一步探讨Eps8调控的PI3K/AKT通路和细胞自噬在逆转TKIs耐药中的作用；为阐明Eps8调控TKIs耐药提供理论和实验依据,为逆转TKIs耐药提供新靶点。

The resistance to tyrosine kinase inhibitors(TKIs) in leukemia patients has restricted its clinical application. Our previously study of bone marrow samples from leukemia patients showed a reverse relationship between the expression level of epidermal growth factor receptor pathway substrate 8(Eps8) and the CR rate of the leukemia patients after one course of chemotherapy, suggested that Eps8 could be involved in the occurrence of drug resistance of leukemia. Eps8 is a new tumor antigen, which involves in the oncogenesis and progression of many solid tumors, and it prompts a bad prognosis. The pre-B lymphocytes leukemic cell line with abnormal Abl genes have been successfully constructed in our former work, which resists to TKIs STI571 and highly expresses Eps8, and as a good model for researching the drug resistance of TKIs in leukemia. In this study, we choose this cell model as the target cells, silence the expression of Eps8 genes by shRNA technology to observe the reversal effect to TKIs, and furtherly investigate the changes of PI3K/AKT pathway and autophagy controlled by Eps8. The research results will provide a theoretical and experimental basis for studying the roles of Eps8 in the resistance of TKIs, and obtain a new target for the reversal of TKIs resistance in leukemic cells.