ENHANCED APOPTOSIS BY TARGETING PROTEIN REDOX STATUS

抗癌新药艾罗富芬和奥沙利铂在肿瘤中是一致和有效的细胞凋亡诱导剂 细胞。伊罗富芬和奥沙利铂可与细胞DNA和蛋白质巯基结合。虽然两者都是DMA 加合物和蛋白质加合物是重要的细胞凋亡刺激物，是蛋白质的潜在致命性后果 艾罗富芬和奥沙利铂的反应性仍未得到充分研究。 这项建议的目的是检验伊罗富文和奥沙利铂靶向关键的假设 氧化还原控制蛋白和随后细胞蛋白氧化还原状态的扭曲增强 癌细胞中的细胞凋亡。蛋白质氧化还原动态平衡主要受抗细胞凋亡的控制 硫氧还蛋白(TRX)系统。Trx被认为是治疗干预的潜在靶点，因为许多 肿瘤通过提高其TRX水平获得生长优势。肿瘤细胞不断处于异常状态 在氧化条件下，可能对阻碍Trx功能的药物特别敏感。 初步数据表明，TRX系统与艾罗富芬和奥沙利铂的作用有关。 第一个目的是研究艾罗富芬和奥沙利铂与Trx和其他关键的氧化还原调节的结合。 纯化形式的蛋白质和癌细胞中的蛋白质。第二个目标是确定伊罗富文的疗效。 和奥沙利铂对细胞氧化还原蛋白的功能和细胞氧化还原状态的扭曲。 全球细胞氧化还原平衡的调制将在所演示的 正常细胞对伊罗富芬和奥沙利铂的耐药性。第三个目标是建立一种因果关系 靶向Trx系统、蛋白质氧化还原状态的全球扭曲和诱导细胞凋亡 伊罗富文和奥沙利铂。这将包括评估氧化还原依赖的ASK1介导的和 低氧条件下依赖TRX的HIF-1信号转导途径。重点是 前列腺癌模型，这与这些药物的临床特性高度相关。成功 完成这些目标将确定TRX系统作为伊罗富文和奥沙利铂靶标的作用， 可以显著改善这些药物的临床使用情况。归根结底，结果应该建立一个 将氧化还原控制蛋白靶向与其他特定抗癌药物相结合的原理 最大限度地提高对DNA损伤剂具有抵抗力的癌细胞的凋亡率。

The novel anticancer drugs irofulven and oxaliplatin are consistent and potent apoptosis inducers in tumor cells. Irofulven and oxaliplatin bind to cellular DNA as well as to protein sulfhydryls. While both DMA adducts and protein adducts are important apoptotic stimuli, the potentially lethal consequences of protein reactivity of irofulven and oxaliplatin remain understudied. The purpose of this proposal is to test the hypothesis that irofulven and oxaliplatin target key redox-controlling proteins and the subsequent distortion of the redox status of cellular proteins enhances apoptosis in cancer cells. Protein redox homeostasis is predominantly controlled by the anti-apoptotic thioredoxin (Trx) system. Trx is recognized as a potential target for therapeutic intervention, as many tumors gain growth advantage by elevating their Trx levels. Tumor cells are constantly under abnormally oxidative conditions and, therefore, may be particularly sensitive to drugs impeding the functions of Trx. Preliminary data implicate the Trx system in the effects of irofulven and oxaliplatin. The first aim is to investigate the binding of irofulven and oxaliplatin to Trx and other key redox regulating proteins in their purified forms and in cancer cells. The second aim is to determine the effects of irofulven and oxaliplatin on the functions of cellular redox proteins and the distortion of the redox status of the cell. Modulation of the global cellular redox balance will be examined in the context of the demonstrated resistance of normal cells to irofulven and oxaliplatin. The third aim is to establish a causative link between the targeting of the Trx system, the global distortion of protein redox status, and the induction of apoptosis by irofulven and oxaliplatin. This will include assessing the roles of redox-dependent ASK1 -mediated and AlF-mediated pathways, as well as Trx-dependent HIF-1 signaling under hypoxic conditions. The focus is on prostate cancer models, which are highly relevant to the clinical properties of these drugs. Successful completion of these aims will define the role of the Trx system as a target for irofulven and oxaliplatin, which could significantly improve the clinical use of these drugs. Ultimately, the outcomes should establish a rationale for combining targeting of redox controlling proteins with other specific anti-cancer drugs to maximize apoptosis in cancer cells that are resistant to DNA damaging agents.