HEAT SHOCK PROTEIN FUNCTION AS A THERAPEUTIC TARGET

描述：调查人员最近的研究表明， 格尔达霉素(GA)与特定细胞热的选择性相互作用 休克蛋白在介导抗肿瘤活性中的作用 这种相互作用在临床相关肿瘤中的分子后果 车型仍不清楚。在这一应用中，提出了以下研究 将检查GA对两个热休克的组装和功能的影响 已知的含有蛋白质的多分子络合物在生物化学中具有重要作用 许多乳腺癌的生物学；即雌激素受体(ER)和 突变型p53。用网织红细胞裂解液Will进行重建实验 用于鉴定ER和P53复合体的特定成分 并评估药物对复杂组装动力学的影响。在 全细胞水平，GA对稳态表达的影响，亚细胞 两者的定位和转录反式激活活性 目标将被检查。最后，检测P53和ER的表达 在GA处理的SCID小鼠体内生长的乳腺肿瘤异种移植瘤中。这些 实验应该为药物作用定义有用的生物终点 活体并将允许评估分子的意义 组织培养对动物致瘤性影响的观察。

DESCRIPTION: The investigator's recent studies have implicated the selective interaction of geldanamycin (GA) with certain cellular heat shock proteins in mediating anti-tumor activity, but the specific molecular consequences of this interaction in clinically relevant tumor models remain unclear. In this application, studies are proposed which will examine GA effects on the assembly and function of two heat shock protein-containing multimolecular complexes known to be important in the biology of many breast cancers; namely, the estrogen receptor (ER) and mutant p53. Reconstitution experiments using reticulocyte lysate will be used to identify the specific components of the ER and p53 complexes and assess drug effects on the dynamics of complex assembly. At the whole-cell level, GA effects on steady state-expression, subcellular localization and transcriptional transactivating activity of the two targets will be examined. Lastly, p53 and ER expression will be examined in breast tumor xenografts growing in SCID mice treated with GA. These experiments should define useful biologic endpoints for drug action in vivo and will allow assessment of the significance of the molecular effects observed in tissue culture for tumorigenicity in animals.