NF-KB PROTEINS AND CELL SURVIVAL

描述：（改编自研究者摘要） 研究NF- B/Rel蛋白在细胞存活中的作用。 我打算利用最近产生的NF-kB缺陷小鼠， IkB RelA亚基在保护性死亡信号中的作用 TNFR家族与癌基因诱导的细胞凋亡的保护作用 1)TNFalpha 对NF-kB缺陷细胞的毒性。 无法执行的基础 RelA-/-巨噬细胞和成纤维细胞在细胞因子的存在下存活。 将研究促炎细胞因子TNF α。 调控 将研究RelA-/-细胞中推定的抗凋亡基因。 的 RelA的特异性结构域和其他NF-κ B亚单位如p50 和c-Rel对TNF α细胞毒性的保护作用将在 RelA-/-、p50-/-RelA-/-和c-Rel-/-RelA-/-细胞。 肿瘤细胞系 将分析对TNF α天然敏感的细胞，以确定NF-kB是否是一种 保护免受TNF α细胞毒性的主要决定因素。 2)作用 RelA保护Fas和TNFR 2细胞毒性。 的灵敏度 RelA-/-T淋巴细胞对Fas和TNFR 2介导的细胞死亡的反应将被抑制。 研究了 这些研究将确定RelA是否在 这些细胞内的抗凋亡或促凋亡能力。 神经酰胺 已经提出在TNF α和Fas-配体刺激后产生， 介导其凋亡作用。 这将由以下人员直接测试： 测定RelA-/-细胞对神经酰胺的敏感性。 3)表达 在RelA-/-成纤维细胞中转化癌基因。 将开展研究， 为了确定致癌性ras或src的能力降低的基础， 转化RelA-/-3T3细胞。 特别是，将进行实验 以确定这些癌基因是否诱导RelA-/-成纤维细胞中的细胞死亡。 将RelA重新引入这些细胞中，以测试是否原位存在RelA。 这种蛋白质是转化和/或防止细胞死亡所必需的。

DESCRIPTION: (adapted from the investigator's abstract) The aim of this investigation is to study the role of NF- B/Rel proteins in cell survival. I intend to take advantage of recently generated mice deficient in NF-kB and IkB RelA subunit in protection death signals generated by members of the TNFR family and in protection from oncogene-induced apoptosis. 1) TNFalpha toxicity to NF-kB deficient cells. The basis for the inability of RelA-/-macrophages and fibroblasts to survive in the presence of the proinflammatory cytokine TNFalpha will be investigated. The regulation of putative anti-apoptotic genes in RelA-/- cells will be studies. The specific domains of RelA and the role of other NF-kB subunits such as p50 and c-Rel in protection from TNFalpha cytotoxicity will be investigated in RelA-/-, p50-/-RelA-/-, and c-Rel-/-RelA-/- cells. Tumor cell lines naturally sensitive to TNFalpha will be analyzed to determine if NF-kB is a primary determinant of protection from TNFalpha cytotoxicity. 2) Role of RelA in protection from Fas and TNFR2 cytotoxicity. The sensitivity of RelA-/-T lymphocytes to Fas and TNFR2 mediated cell death will be investigated. These studies will determine whether RelA functions in an anti-apoptotic or pro-apoptotic capacity within these cells. Ceramide generated following TNFalpha and Fas-ligand stimulation has been proposed to mediate their apoptotic affects. This will be directly tested by determining the sensitivity of RelA-/- cells to ceramide. 3) Expression of transforming oncogenes in RelA-/- fibroblasts. Studies will be carried out to determine the basis for decreased capability of oncogenic ras or src to transform RelA-/-3T3 cells. In particular, experiments will be carried out to determine if these oncogenes induce cell death in RelA-/-fibroblasts. RelA will be reintroduced in these cells to test if the in situ presence of this protein is required for transformation and/or preventing cell death.