CLONING OF GENES INVOLVED IN CELL IMMORTALIZATION

描述：麦考密克博士正在对这些变化的性质进行因果关系研究 参与将正常人类细胞转化为癌细胞。强壮 有证据表明，避免衰老是 涉及到的变化。他们成功地创造了两个无限的寿命 人成纤维细胞株MSU-1.0和MSU-2，来源于 转v-myc基因的克隆化群体。他们和其他人 有数据表明表达上调或放松监管 Myc基因的表达有助于人类永生 间充质细胞，但不足以让它们从 衰老。细胞还必须失去一个基因的两个等位基因的活性(S) 这就防止了这样的逃脱。这项提案的目的是确定和 鉴定这种myc互补基因或其活性必须是 这些细胞失去了长生不老的能力。他们一直在检查这件事 二倍体菌株MSU-1.0的基因差异显示问题 并发现有大量下调的基因 与其亲本LG1细胞中的表达水平进行比较。他们 将测试这样确定的候选基因是否具有返回 MSU 1.0细胞进入致死状态。如果这些研究在 确定感兴趣的基因(S)，他们将检查 在这些细胞中表达上调。尽管对数据的通常解释是 细胞融合研究的最新进展是永生化涉及下调 基因，人们可以建立基因表达上调的模型， 例如端粒酶基因。如果他们没有成功地识别出 在与MSU 1.0细胞的研究中，他们将携带MYC互补基因 Out DD在另外两个不朽的人成纤维细胞系(MSU-2和MSU-2)上的表达 OUMS-24F)和它们各自的同基因致死亲本细胞(SL80和SL80 OUMS)。人类肉瘤以及人类肉瘤来源的细胞系， 经常表现为myc表达上调。30株人肉瘤细胞系 在他们的实验室里都有。增加……成功的机会 确定感兴趣的基因(S)，他们将包括在他们的分析 MSU-2和OUMS-24F细胞表达水平的分析 在这些人类肉瘤来源的细胞系中发现的基因，他们鉴定为 过度表达myc。将对DD确定的候选基因进行测试 将细胞恢复到人类状态的能力。

DESCRIPTION: Dr. McCormick is studying the nature of the changes causally involved in transforming normal human cells into cancer cells. Strong evidence indicates that escape from senescence is one of the fundamental changes involved. They have succeeded in generating two infinite life span human fibroblast cell strains, MSU-1.0 and MSU-2, which arose from clonally-derived populations transfected with a v-myc gene. They and others have data indicating that upregulation of expression or deregulation of expression of the myc gene contributes to immortalization of human mesenchymal cells, but is not sufficient to allow them to escape from senescence. Cells must also lose the activity of both alleles of a gene(s) that prevents such escape. The aim of this proposal is to identify and characterize such myc-complementing gene or genes whose activity must be lost for such cells to become immortal. They have been examining this question using mRNA Differential Display (DD) for the diploid strain MSU-1.0 and have found that there are a significant number of down-regulated genes compared with the level of expression in their parental LG1 cells. They will test the candidate genes so identified for their ability to return the MSU 1.0 cells to the mortal state. If these studies are not successful in identifying the gene(s) of interest, they will examine the genes that are up-regulated in these cells. Although the usual interpretation of the data of cell fusion studies is that immortalization involves down-regulation of genes, one can build models in which expression of genes are up-regulated, e.g. the telomerase gene. If they do not succeed in identifying the myc-complementing gene in the studies with MSU 1.0 cells, they will carry out DD on two other immortal human fibroblast cell lines (MSU-2 and OUMS-24F) and their respective isogeneic mortal parental cells (SL80 and OUMS). Human sarcomas, as well as human sarcoma-derived cell lines, frequently exhibit up-regulation of myc. Thirty human sarcoma cell lines are available in their laboratory. To increase the chances of success in identifying the gene(s) of interest, they will include in their analysis of the MSU-2 and OUMS-24F cells, an analysis of the levels of expression of genes found in those human sarcoma derived cell lines that they identify as overexpressing myc. Candidate genes identified by DD will be tested for ability to return the cells to the mortal state.