HEAT AND RADIATION INDUCED CHANGES IN THE NUCLEAR MATRIX

核矩阵对于定义核结构和 调节代谢过程，如DNA和RNA合成，并可能 DNA修复。我们建议确定核矩阵是否是 参与热损伤表现的关键结构和 热辐射增敏。我们将测试假设中的变化 核基质蛋白与热诱导的改变有关(或 核基质结构的恢复与细胞毒性有关。 使用针对特定核基质蛋白的抗体，我们将 定量测定核层粘连蛋白和内部基质的热诱导变化 蛋白质，我们将量化热诱导的核层和核层的变化。 内部基质蛋白，我们将量化热诱导的变化 用流式细胞术原位检测核层粘连蛋白和内基质蛋白 测定细胞内这些蛋白质的相对丰度 周而复始。固定细胞的结构变化将通过共聚焦进行评估 显微镜下观察，并与染色质分布和 生死存亡。因为核矩阵的变化可能会被认为是 有助于在热休克过程中观察到DNA链断裂修复的抑制 放射增敏，我们还将确定时间和空间模式 在受热、受辐射的细胞中进行DNA修复。流式细胞仪定量检测 核基质蛋白的变化可能具有临床意义 对抗癌治疗反应的预测分析，如 体温过高，因为我们的初步数据表明，热导致 几小时内层蛋白B在凋亡坏死细胞中的增溶作用 暖气。核基质蛋白的增溶可能被证明是一种 快速替代集落形成试验预测热敏感度。 热疗单独或联合作用机制的阐明 有了辐射，可能会导致癌症治疗的优化和 优先根除肿瘤细胞。

The nuclear matrix is important for defining nuclear structure and regulating metabolic processes such as DNA and RNA synthesis, and possibly DNA repair. We propose to determine whether the nuclear matrix is a critical structure involved in the manifestation of thermal damage and heat-radiosensitization. We will test the hypotheses that alterations in nuclear matrix proteins are related to heat-induced alterations in (or recovery of) nuclear matrix structure are correlated with cytotoxicity. Using antibodies against specific nucleic matrix proteins, we will quantitate heat-induced changes in nuclear lamin and interior matrix proteins, we will quantitate heat-induced changes in nuclear lamina nd interior matrix proteins, we will quantitate heat-induced changes in nuclear lamin and interior matrix proteins in situ using flow cytometry to determine the relative abundance of these proteins throughout the cell cycle. Structural changes in fixed cells will be evaluated with confocal microscopy and correlated with change in chromatin distribution and survival. Since changes ina the nuclear matrix might be expected to contribute to inhibition of DNA strand break repair observed during heat- radiosensitization, we will also determine temporal and spatial patterns of DNA repair in heated, irradiated cells. Flow cytometric quantitation of changes in nuclear matrix proteins may have clinical relevance as a predictive assay for response to anticancer treatments such as hyperthermia, since our preliminary data suggests that heat causes solubilization of lamin B in apoptotic an necrotic cells within hours after heating. Solubilization of nuclear matrix proteins could prove to be a rapid alternative to colony-forming assays for predicting heat sensitivity. Elucidation of the mechanisms of action of hyperthermia, alone or combined with radiation, may lead to optimization of cancer treatments and the preferential eradication of tumor cells.