IMMORTALIZATION OF SV40 TRANSFORMED HUMAN CELLS

SV40 转化人类细胞的永生化

• 批准号: 2748485
• 负责人: HARVEY L OZER
• 金额: $ 41.66万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2748485
• 关键词: antisense nucleic acid; biological signal transduction; cell senescence; cell transformation; fibroblasts; flow cytometry; gene expression; gene mutation; genetic library; genetic promoter element; laboratory mouse; mitogen activated protein kinase; simian virus 40; tissue /cell culture; tumor suppressor genes; yeast two hybrid system

DESCRIPTION: (Adapted from the applicant's abstract) Normal human cells have a limited lifespan in culture and undergo replicative senescence whereas tumor cells commonly grow continuously and are immortal. Hence cellular senescence also functions in part as growth/tumor suppressor in a particular aspect of carcinogenesis. They have been exploiting SV40-transformed human fibroblasts (SV/HF) as a model to understand the molecular mechanism of immortalization. Utilizing matched sets of non-immortal (preimmortal) and immortal SV/HF, they eliminated changes in viral gene functions as being responsible and, therefore, sought to identify relevant cellular genes including an essential primary effector termed SEN6 (by molecular genetic approaches) and secondary effectors (by differential screening of a cDNA library) in the period of the last grant award. Each approach has resulted in the identification of a gene which had not previously been extensively studied or considered associated with senescence or immortalization. They propose to investigate their effects on normal cells and SV/HF. They have determined that a locus, which is inactivated upon immortalization, maps to 6q27 and have identified a gene in this region which has growth suppressor activity and has been mutated in immortal SV/HF, consistent with being SEN6. In addition, they have isolated a cDNA for a gene whose expression is reduced in immortal SV/HF (as compared to preimmortal SV/HF and normal cells) which has sequences indicative of a protein serine/threonine phosphatase (PSP). Proposed experiments will determine whether each is effective as a growth suppressor in SV/HF and other tumor-derived immortal cell lines, and assess the mechanism involved. Both SEN6 and PSP have properties which indicate that they might act at different stages of a common pathway involving MAP kinases so they will initially focus their attention on changes in those pathways in immortal SV/HF and upon overexpression (under the control of a regulatable promoter) of introduced sequences for the candidate for SEN6 and/or the protein phosphatase. They anticipate that the results will generate new insights into cellular senescence and its role in carcinogenesis.

描述：（改编自申请人摘要）正常人细胞 在培养中寿命有限， 而肿瘤细胞通常连续生长并且是永生的。 因此 细胞衰老也部分地作为生长/肿瘤抑制因子， 致癌作用的特殊方面。 他们一直在利用 SV 40转化的人成纤维细胞（SV/HF）作为模型来理解 永生化的分子机制 利用匹配的 非永生（永生前）和永生SV/HF，他们消除了 病毒基因的功能是负责，因此，试图确定 相关的细胞基因，包括称为SEN 6的必需初级效应子 (by分子遗传学方法）和次级效应物（通过差异 筛选cDNA文库）。 每个 这种方法已经鉴定出一种基因， 以前被广泛研究或认为与衰老有关 或者永生 他们建议研究它们对正常人的影响。 细胞和SV/HF。 他们已经确定，一个位点，这是失活后， 永生化，映射到6 q27，并已确定在该区域的基因 其具有生长抑制活性并已在永生SV/HF中突变， 与SEN 6一致。 此外，他们还分离出了一种 在永生SV/HF中表达降低的基因（与 永生前SV/HF和正常细胞），其具有指示细胞增殖的序列。 蛋白丝氨酸/苏氨酸磷酸酶（PSP）。 拟议的实验将 确定每一种是否都是SV/HF中有效的生长抑制剂， 其他肿瘤来源的永生细胞系，并评估所涉及的机制。 SEN 6和PSP都具有表明它们可能在 不同阶段的共同途径，涉及MAP激酶，所以他们将 最初，他们将注意力集中在不朽的那些途径的变化上， SV/HF和过表达时（在可调控启动子的控制下） SEN 6和/或蛋白质的候选物的引入序列 磷酸酶。 他们预计这些结果将产生新的见解 细胞衰老及其在致癌作用中的作用。