基于化学生物学的耐药白血病治疗新靶点的发现

项目名称： 基于化学生物学的耐药白血病治疗新靶点的发现

项目编号： No.30873091

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 熊冬生

作者单位： 中国医学科学院

项目金额： 34万元

中文摘要： 多药耐药是指肿瘤细胞对一种化疗药物出现耐药的同时,对其他结构不同、作用靶位不同的化疗药物亦产生抗药性，是肿瘤化疗失败和复发的根源。我们以吲哚环为母核合成了其多样性衍生物库，筛选到具有抑制多种多药耐药肿瘤细胞和其亲代药物敏感细胞的新化合物PH II-7，以PH II-7为探针，从K562/A02细胞中钓取出两个与PH II-7相互作用的蛋白，并在分析PHII-7作用前后多药耐药K562/A02 及其亲代K562细胞基因表达谱差异的基础上，证明PH II-7通过作用于细胞周期相关基因和凋亡相关基因导致K562/A02 和K562细胞阻滞在S期，并诱导细胞凋亡；证明PH II-7通过PKCA下调MDR1基因的表达，从而逆转K562/A02 细胞的耐药性；在国内外首次发现sorcin 基因的表达与临床白血病耐药密切相关。证明PH II-7下调多药耐药HL-60/ADR细胞MRP基因的表达，从而逆转其对ADR的耐药性。首次发现ABCB5在白血病细胞表达，并与临床患者耐药密切相关。首次构建了IL-3与LDM融合蛋白，证明其具有靶向杀伤CD123阳性肿瘤细胞的作用。该研究为耐药白血病的治疗奠定了基础。

中文关键词： 白血病;耐药;基因表达谱;凋亡;细胞周期

英文摘要： Although the concepts and technologies used for cancer therapy have evolved greatly, most clinical cancer treatments still rely on chemotherapy, especially for metastatic cancer and hematological malignancies. However, tumor cells treated with chemotherapeutic agents commonly develop resistance to a variety of structurally and mechanistically unrelated drugs; this phenomenon is called multi-drug resistance (MDR),Which is the leading cause of chemotherapy false and relapse. Using indole ring as the parent nucleus,we synthesized its diversity derivative library and screened out the new anti-tumor agent PH II-7, which inhibited several MDR tumor cell lines and their parent drug-sensitive cell lines. Using PH II-7 as a probe, two proteins intereacted with PH II-7 were baited from K562/A02 cell line. Meanwhile，we analyzed the genome profiling changes in PH II-7 treated K562 and K562/A02 cells and found that PH II-7 can induce apoptosis and S phase cell cycle arrest by up or down-regulating the expression of cell cycle or apoptosis-associated genes and effectively inhibits the expression of MDR1 and impairs the drug efflux function of P-gp by reducing the expression level of PKCA. We discovered that the over expression of sorcin is associated with a MDR human leukemia cell line K562/A02 induced by DOX, and for the first time discovered the significant association between the sorcin over expression and drug resistance in leukemia in clinic.We also confirmed that PH II-7 down-regulated MRP and significantly sensitized HL-60/ADR cells to chemotherapeutic drugs mediated cytotoxicity. We discovered that the overexpression of ABCB5 may be responsible both for the progression and chemotherapeutic refractoriness of advanced acute leukemia.Additionally, we slao constructed and expressed IL3-LDM fusion protein, and found that IL3-LDM exerted potent cytotoxicity against CD123+ cell lines in vitro.The study lay the experimental foundationg for treatment of drug resistance in leukemia.

英文关键词： leukemia;drug resistance;cell cycle; genome profiling;apoptosis