Affiliation:
1. Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark 07103-2714.
Abstract
Human diploid fibroblasts have a limited life span in vitro, and spontaneous immortalization is an extremely rare event. We have used transformation of human diploid fibroblasts by an origin-defective simian virus 40 genome to develop series of genetically matched immortal cell lines to analyze immortalization. Comparison of a preimmortal transformant (SVtsA/HF-A) with its uncloned and cloned immortalized derivatives (AR5 and HAL) has failed to reveal any major alteration involving the simian virus 40 genome. Karyotypic analysis, however, demonstrated that all of the immortal cell lines in this series have alterations of chromosome 6 involving loss of the portion distal to 6q21. The karyotypic analysis was corroborated by DNA analyses. Southern analysis demonstrated that only one copy of three proto-oncogene loci (ros1, c-myb, and mas1) on 6q was retained in immortal cells. Polymerase chain reaction analysis of the microsatellite polymorphism at 6q22 (D6S87) showed loss of heterozygosity. In addition, elevated expression of c-myb (6q22-23) was observed. We hypothesize that the region at and/or distal to 6q21 plays a role in immortalization, consistent with the presence of a growth suppressor gene.
Publisher
American Society for Microbiology
Subject
Cell Biology,Molecular Biology
Reference66 articles.
1. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA;Auffrey C.;Eur. J. Biochem.,1980
2. Ausubel F. M. R. Brent R. E. Kingston D. D. Moore J. G. Seidman J. A. Smith and K. Struhl (ed.). 1987. Current protocols in molecular biology. Greene Publishing Associates and Wiley-Interscience New York.
3. Structure of two forms of the interferon-induced (2'-5') oligoA synthetase of human cells based on cDNAs and gene sequences;Benech P.;EMBO J.,1985
4. Expression and rearrangement of the ROSI gene in human glioblastoma cells;Birchmeier C.;Proc. Natl. Acad. Sci. USA,1987
5. Spontaneous abnormalities in normal fibroblasts from patients with Li-Fraumeni cancer syndrome: aneuploidy and immortalization;Bischoff F. Z.;Cancer Res.,1990
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献