CLONING OF GENES INVOLVED IN CELL IMMORTALIZATION

描述：麦考密克博士正在研究因果关系变化的性质 参与将正常人体细胞转化为癌细胞。 强 有证据表明，逃避衰老是一个基本的 涉及的变化。 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，其来源于 用V-myc基因转染的克隆衍生群体。 他们和其他人 有数据表明，表达的上调或失调， myc基因的表达有助于人类的永生化 间充质细胞，但不足以让他们逃脱， 衰老 细胞也必须失去一个基因的两个等位基因的活性 防止这种逃跑。 这项建议的目的是确定和 表征这样的一个或多个myc互补基因，其活性必须 这些细胞无法永生 他们一直在研究 二倍体菌株MSU-1.0的mRNA差异显示（DD）问题 并发现有相当数量的下调基因 与其亲本LG 1细胞中的表达水平相比。 他们 将测试这样确定的候选基因的能力， MSU 1.0细胞到死亡状态。 如果这些研究不成功， 识别感兴趣的基因，他们将检查基因， 在这些细胞中表达上调。 虽然通常对数据的解释 细胞融合研究的一个重要结论是，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 如果他们不能成功地识别出 在MSU 1.0细胞的研究中，它们将携带myc互补基因， 在另外两种永生人成纤维细胞系（MSU-2和 OUMS-24 F）和它们各自的同基因致死亲本细胞（SL 80和SL 84）。 OUMS）。 人肉瘤以及人肉瘤衍生的细胞系， 经常表现出myc的上调。 30种人肉瘤细胞系 在他们的实验室里。 为了增加成功的机会 鉴定感兴趣的基因，他们将包括在他们的分析， MSU-2和OUMS-24 F细胞中，分析了 在那些人类肉瘤衍生细胞系中发现的基因， 过表达myc. 将检测通过DD鉴定的候选基因， 使细胞恢复到正常状态的能力。

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.