Proteomic and Functional Studies of Mitochondrial Proteins Involved in ROS Metabo

描述(申请人提供)：导致胰腺癌病死率高的主要因素之一是大多数胰腺癌患者对包括放射治疗和化疗在内的治疗反应不佳。我们这个项目的长期目标是了解胰腺癌耐药的分子机制。诱导靶细胞的凋亡是放化疗诱导细胞杀伤的关键机制。线粒体和线粒体产生的活性氧(ROS)在细胞死亡途径中起着重要作用。在线粒体依赖的凋亡途径中，在细胞色素c等细胞色素c等凋亡因子释放后，下游的生化途径似乎被不同的细胞所共享。我们推测，参与产生或清除ROS的线粒体蛋白决定了线粒体中的ROS水平，可能是导致胰腺细胞对治疗产生抵抗力的原因。因此，我们建议使用定量蛋白质组学方法系统地筛选对ROS诱导药物敏感性不同的胰腺癌细胞中参与ROS产生和清除的线粒体蛋白(特定目标1)。在蛋白质组筛选后，将使用RNAi或过表达技术来操纵已鉴定的目标蛋白在细胞中的表达，并将确定目标蛋白的表达变化对细胞ROS积累和细胞凋亡的影响(特异性目标2)。识别与胰腺癌细胞对抗癌药物耐药密切相关的蛋白质因子，可能为设计新的治疗策略以克服胰腺癌对药物的耐药性提供有价值的信息。 公共卫生相关性：该项目的目标是确定可能导致胰腺癌化疗耐药的新的蛋白质靶点。新的定量蛋白质组学方法将被用来系统地比较化疗敏感和耐药胰腺癌细胞中的线粒体氧化还原蛋白。分子和细胞生物学技术将被用来评估识别的目标蛋白的生物学功能。

DESCRIPTION (provided by applicant): One of the major factors contributing to the high fatality of pancreatic cancer is the poor response of most pancreatic cancer patients to therapies including radiation therapy and chemotherapy. Our long- term objective of this project is to understand the molecular mechanisms underlying the resistance of pancreatic cancer to therapies. Induction of apoptosis in target cells is a key mechanism by which radiation and chemotherapy induces cell killing. Mitochondrion and mitochondrion-generated reactive oxygen species (ROS) play essential roles in the cell death pathways. In the mitochondria dependent apoptotic pathways, after the release of the apoptotic factors, such as cytochrome c, the downstream biochemical pathways appear to be shared by different cells. We hypothesize that the mitochondrial proteins involved in ROS production or scavenging, which determine the ROS levels in mitochondria, may be responsible for the resistance of pancreatic cells to therapies. Accordingly, we propose to use quantitative proteomic methods to systematically screen mitochondrial proteins that are involved in ROS production and scavenging in pancreatic cancer cells with different sensitivity to ROS inducing drugs (Specific Aim 1). After the proteomic screening, RNAi or overexpression techniques will be used to manipulate the expression of the identified target proteins in cells, and the effects of the altered expression of the target proteins on cellular ROS accumulation, and cellular apoptosis will be determined (Specific Aim 2). Identifications of the protein factors that are closely associated with the resistance of pancreatic cancer cells to anti-cancer drugs may provide valuable information for designing new novel therapeutic strategies to overcome the resistance of pancreatic cancer to therapies. PUBLIC HEALTH RELEVANCE: The objective of this project is to identify novel protein targets that may be responsible for chemo- resistance of pancreatic cancer. Novel quantitative proteomic methods will be used to systemically compare mitochondrial redox proteins in chemo-sensitive and chemo-resistant pancreatic cancer cells. Molecular and cell biological techniques will be used to assess the biological functions of the identified target proteins.