MOLECULAR AND CYTOGENETIC STUDIES OF HUMAN CELL AGING

人类细胞衰老的分子和细胞遗传学研究

• 批准号: 6471199
• 负责人: OLIVIA M. PEREIRA-SMITH
• 金额: $ 33.78万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6471199
• 关键词: HeLa cells; cell age; cell bank /registry; cell cycle; cell growth regulation; cell line; cell population study; cell senescence; cell transformation; chromosomes; clone cells; cytogenetics; gene expression; genetic library; genetic manipulation; genetic mapping; human tissue; hybrid cells; molecular cloning; monoclonal antibody; neoplasm /cancer genetics; nucleic acid probes; nucleic acid sequence; polymerase chain reaction; southern blotting

Normal human cells in vitro exhibit a stringent limitation of division capacity. In contrast, tumor-derived and virus-, carcinogen- or irradiation-transformed cells can divide indefinitely (immortal). We do not yet understand the mechanisms that limit the division potential of normal human cells or the changes occurring to yield immortal cells. However, from cell hybrid studies we have found that the immortal phenotype results from recessive changes in normal cell growth control. We have exploited this fact to separate twenty four different immortal cell lines into four complementation groups for indefinite division. This indicates that there are at least four paths to immortality and that at least four genes or sets of genes are involved in normal cell growth control. Using the technique of microcell mediated chromosome transfer, we have determined that the introduction of a normal human chromosome 4 into three immortal human cell lines that assign to complementation group B, results in loss of proliferation. This is a specific effect to the group B cell lines, because the division capability of immortal cell lines assigned to the other groups is not affected by this chromosome, and a normal chromosome 11 has no effect on any of the immortal cell lines tested. These results indicate that there is a cell senescence related gene(s) on human chromosome 4. We have also found that a normal chromosome 1 induces loss of proliferation in one immortal human cell line that assigns to group C., In this proposal, we plan to take multipronged approaches to clone the gene on chromosome 4 and study regulation of its expression during senescence and immortalization. We will also determine whether chromosome 1 affects the proliferation potential of other immortal human cell lines that assign to group C. If it does, this would indicate that another cellular senescence gene(s) is present on this chromosome and we will attempt to localize the region on chromosome 1 that is involved.

正常的人类细胞体外表现出严格的分裂限制 容量。 相反，肿瘤衍生和病毒，致癌或 辐照转换的细胞可以无限期分裂（不朽）。 我们做 尚不了解限制分裂潜力的机制 正常的人类细胞或发生的变化以产生不朽的细胞。 但是，从细胞杂种研究中，我们发现不朽 表型是由于正常细胞生长控制的隐性变化而导致的。 我们已经利用了这一事实将二十四个不同的不朽 细胞系分为四个不确定分裂的互补组。 这表明至少有四个永生途径， 至少四个基因或一组基因参与正常细胞生长 控制。 使用Microcell介导的染色体转移技术， 我们已经确定引入正常的人类染色体4 分为分配给互补组的三种不朽的人类细胞系 B，导致增殖的丧失。 这是对 B组细胞系，因为不朽细胞的分裂能力 分配给其他组的线不受此染色体的影响， 正常染色体11对任何不朽的细胞没有影响 测试线。 这些结果表明有一个细胞衰老 人4染色体上的相关基因。我们还发现正常 染色体1诱导一个不朽的人类细胞中的增殖丧失 分配给C组的行，我们计划采取 多收集的方法在4号染色体上克隆基因并研究 调节其在衰老和永生化过程中的表达。 我们 还将确定1染色体是否影响增殖 分配给组的其他不朽的人类细胞系的潜力 确实，这表明另一个细胞衰老基因是 在此染色体上存在，我们将尝试将该区域定位在 涉及的染色体1。