两种小分子化合物抗癌作用的发现与机制研究

发布时间：2018-06-05 01:46

本文选题：DNA损伤反应 + 5-碘代杀结核菌素　；参考：《上海交通大学》2014年博士论文

【摘要】：许多化疗药物是基因毒性药物,通过激活DNA损伤应答来抑制细胞增殖或诱导细胞死亡。p53是一种重要的抑癌蛋白,也是抗癌药物研发的重要靶点。我们以是否激活p53为指标,筛选激酶和磷酸酶抑制剂寻找新化疗药物。发现嘌呤衍生物5-Iodotubercidin(Itu,腺苷激酶抑制剂)和双吲哚马来酰亚胺家族衍生物Bisindolylmaleimide IX(蛋白激酶C抑制剂)具有激活p53的作用。第一部分,我们在几个细胞系中证实了Itu可诱导p53的表达。我们发现它会导致DNA损伤,诱导DNA断裂,产生γH2AX和TopBP1灶,激活Atm、Chk2、p53。虽然作为腺嘌呤类似物的Itu是腺苷激酶(ADK)的抑制剂,但我们证明它激活p53与抑制ADK的活性无关。Itu以p53依赖的方式使细胞周期阻滞在G2/M期,以p53依赖和非依赖性两种方式诱导细胞死亡。DNA断裂可能是由于Itu代谢产物的掺入所引起。Itu显示抗肿瘤活性,以p53依赖和非依赖性方式抑制裸鼠异种移植模型中的肿瘤的生长。第二部分,通过比较Bisindolylmaleimide IX对多种肿瘤细胞系的杀伤效果,发现它具有治疗慢性粒细胞白血病(CML,包括伊马替尼药物抗性)的潜力。BCR-ABL抑制剂治疗CML往往由于耐药性的出现使其效果欠佳。我们发现Bisindolylmaleimide IX是一种DNA拓扑异构酶抑制剂,激活DNA损伤反应。相比于其它测试的癌细胞系,它对BCR-ABL阳性细胞靶向作用好,而BCR-ABL是高效诱导基因组不稳定的激酶。Bisindolylmaleimide IX对BCR-ABL阳性细胞的DNA损伤较强,减少其拓扑异构酶的表达,使细胞周期停滞于G2/M期。同时它也抑制Raf和BCR-ABL(包括耐药突变T315I的BCR-ABL)下游成瘾途径的癌基因。我们的实验证明Bisindolylmaleimide IX也是双吲哚马来酰亚胺家族众多衍生物中最有效的一个。裸鼠肿瘤模型实验表明,它有效抑制BCR-ABL(包括T315I耐药突变)诱导的肿瘤的生长,延长CML模型裸鼠的生存期。Bisindolylmaleimide IX通过激活BCR-ABL依赖的基因毒性应激反应和抑制成瘾途径相关癌基因来治疗包括具有药物抗性的CML,是一种有潜力的治疗药物。
[Abstract]:Many chemotherapeutic drugs are genotoxic drugs. By activating DNA damage response to inhibit cell proliferation or induce cell death. P53 is an important tumor suppressor protein and an important target for the development of anticancer drugs. We looked for new chemotherapeutic agents by screening kinase and phosphatase inhibitors based on whether p53 was activated. It was found that the purine derivative 5-Iodocidinine (adenosine kinase inhibitor) and the diindole maleimide family derivative Bisindolylmaleimide IX (protein kinase C inhibitor) could activate p53. In the first part, we confirmed that Itu could induce p53 expression in several cell lines. We found that it could cause DNA damage, induce DNA breakage, produce 纬 H2AX and TopBP1 foci, and activate Atmna Chk2P53. Although Itu, an adenine analogue, is an inhibitor of adenosine kinase (ADK), we have shown that its activation of p53 has nothing to do with inhibiting the activity of ADK. ITU blocks the cell cycle in G 2 / M phase in a p53 dependent manner. Both p53 dependent and non-dependent inducing cell death. DNA breakage may be due to the incorporation of Itu metabolites, and inhibit tumor growth in nude mice xenotransplantation model by p53 dependent and independent manner. In the second part, by comparing the killing effect of Bisindolylmaleimide IX on various tumor cell lines, It was found that BCR-ABL inhibitor has the potential to treat chronic myeloid leukemia (CML), including imatinib drug resistance. BCR-ABL inhibitors often have poor efficacy in the treatment of CML due to the emergence of drug resistance. We found that Bisindolylmaleimide IX is a DNA topoisomerase inhibitor that activates the DNA damage response. Compared with other cancer cell lines tested, it has a good targeting effect on BCR-ABL positive cells, while BCR-ABL is a highly genome-induced genomic unstable kinase. Bisindolylmaleimide IX has stronger DNA damage and reduces the expression of topoisomerase in BCR-ABL positive cells. Cell cycle arrest at G 2 / M phase. It also inhibits downstream addictive pathway oncogenes of Raf and BCR-ABL (including BCR-ABL with drug-resistant mutation T315I). Our experiments show that Bisindolylmaleimide IX is also the most effective one of the many derivatives of the indole-maleimide family. The tumor model of nude mice showed that BCR-ABL (including T315I resistant mutation) induced tumor growth. Prolonging the survival of nude mice with CML. Bisindolylmaleimide IX is a potential therapeutic drug by activating BCR-ABL dependent genotoxic stress response and inhibiting addictive pathway related oncogenes including drug-resistant CMLs.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R96

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