HEAT SHOCK PROTEINS AND CELL RESISTANCE TO HYPERTHERMIA

热休克蛋白和细胞对高温的抵抗力

• 批准号: 3302137
• 负责人: LEE A WEBER
• 金额: $ 12.93万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3302137
• 关键词: DNA repair; RNA biosynthesis; autoradiography; cell death; cytoskeleton; densitometry; fluorescence microscopy; gel electrophoresis; gene expression; genetic transcription; genetic translation; hamsters; human tissue; hybrid cells; hyperthermia therapy; laboratory mouse; molecular site; neoplasm /cancer therapy; nucleic acid hybridization; phosphorylation; protein biosynthesis; protein metabolism; scintillation counter; site directed mutagenesis; stress proteins; tissue /cell culture; transfection

The goal of this study is to understand how cells resist killing by heat and how they develop increased levels of thermotolerance after surviving hypothermia. In a collaborative project between the Weber and Landry laboratories, we have found that human gene encoding the 27 kDa heat shock protein is expressed constitutively in rodent cells and produces a heat resistant phenotype. Using specific antibody to the human hsp27 protein, we find a good correlation between the amount of human hsp27 in stably transformed hamster and mouse cell lines and the degree of thermal resistance. These cell lines will be used in the present study to learn whether overexpression of hsp27 in the absence of the other heat shock proteins can inhibit formation of accelerate the repair of specific lesions in cell function and structure that are known to be caused by heat. These include inhibition of transcription, RNA processing, translation, increased protein catabolism, and alterations in the structure of the cytoskeleton. We will also investigate the importance of phosphorylation of he hsp27 protein in determining thermal resistance in response to thermoprotecting and thermosensitizing agents. The sites of phosphorylation in the protein will be determined and the significance of each site for thermoprotection following heat shock will be evaluated by site-directed mutagenesis studies. Finally, the high constitutive activity of the human hsp27 gene in rodent cells will be used to develop transformed cell lines that overexpress other genes and cDNAs encoding different heat shock proteins. The phenotype of each type cell line will be studied with respect to thermoresistance and protection from known heat induced lesions. Cell lines that overexpress multiple heat shock proteins will also be developed to test for functional interactions between the different proteins. Information obtained from this study will provide a better understanding of how cells respond the hyperthermia which could lead to more rational approaches to thermotherapy in cancer treatment.

这项研究的目的是了解细胞如何抵抗热杀死 以及它们在存活后如何提高耐热性 体温过低 在韦伯和兰德里的一个合作项目中 在实验室中，我们发现编码27 kDa热休克的人类基因 蛋白质在啮齿类动物细胞中组成型表达， 抗性表型 利用针对人hsp27蛋白的特异性抗体， 我们发现稳定表达的人HSP 27的量与稳定表达的人HSP 27的量之间有很好的相关性。 转化的仓鼠和小鼠细胞系和热转化的程度 阻力 这些细胞系将用于本研究，以了解 是否在没有其他热休克的情况下过度表达HSP 27 蛋白质可以抑制特定损伤的形成， 在细胞功能和结构中的作用。 这些 包括抑制转录、RNA加工、翻译、增加 蛋白质催化剂和细胞骨架结构的改变。 我们还将研究hsp27磷酸化的重要性。 蛋白质在确定响应于热保护的热阻中的作用 和热敏剂。 蛋白质中磷酸化的位点 将确定和意义，每个网站的热防护 热休克后将通过定点诱变进行评价 问题研究 最后，人hsp27基因的高组成活性 将用于开发转化细胞系， 过表达其他基因和编码不同热休克蛋白的cDNA。 将研究每种类型细胞系的表型， 耐热性和对已知热诱导损伤的保护。 细胞 还将开发过表达多种热休克蛋白的品系 来测试不同蛋白质之间的功能性相互作用。 从这项研究中获得的信息将有助于更好地了解 细胞如何对高温做出反应， 癌症热疗的方法。