Journal article
Immortalization in a normal foreskin fibroblast culture following transduction of cyclin A2 or cdk1 genes in retroviral vectors
Experimental cell research, Vol.294(2), pp.406-419
2004
DOI: 10.1016/j.yexcr.2003.11.030
PMID: 15023530
Abstract
Human diploid fibroblasts (HDF) rarely, if ever, undergo spontaneous transformation to an immortalized cell type. Here we report the immortalization of an HDF cell line following transduction with
cyclin A2 or
cdk1 human genes via retroviral vectors. Fluorescence in situ hybridization (FISH) studies using the retroviral vector as a probe indicate that these cell lines are monoclonal. No telomerase activity could be detected in these cell lines, and the telomere length in the immortalized cells was observed to be 10–20 kb longer than that in low-passage cells from the parental fibroblast line. Cytogenetic studies revealed that the immortal lines share common chromosomal aberrations. FISH studies with a probe for
p53 revealed loss of one copy of this gene which was associated with reduced steady-state levels of both
p53 and
p53-regulated
p21
WAF1/Sdi1/CIP1
messages in both quiescent and proliferating immortalized cultures relative to the parental cells. Additional FISH studies with probes for
p16
INK4a
and
Rb, carried out after the immortalized cells proliferated in excess of 100 population doublings, also revealed loss of one copy of these genes in both cell lines. These cell lines, together with the well-characterized parental cells, could provide useful research material for the study of the mechanisms of immortalization and of regulation of proliferative senescence in HDF.
Details
- Title: Subtitle
- Immortalization in a normal foreskin fibroblast culture following transduction of cyclin A2 or cdk1 genes in retroviral vectors
- Creators
- Ping Luo - Department of Pathology, University of Washington Seattle, Seattle, WA 98195, USAMaria Tresini - Lankenau Medical Research Center, Wynnewood, PA 19096, USAVincent Cristofalo - Lankenau Medical Research Center, Wynnewood, PA 19096, USAXiaoqin Chen - Department of Pathology, University of Washington Seattle, Seattle, WA 98195, USAAnthony Saulewicz - Department of Pathology, University of Washington Seattle, Seattle, WA 98195, USAMatthew D Gray - Combimatrix Corp., Mulkiteo, WA 98275, USADeborah E Banker - Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USAAloysius L Klingelhutz - Department of Microbiology, University of Iowa, Iowa City, IA 52247, USAMotoaki Ohtsubo - Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima, 1-2-3 Kasumi, Minami, Hiroshima 734-8553, JapanYoshihiro Takihara - Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima, 1-2-3 Kasumi, Minami, Hiroshima 734-8553, JapanThomas H Norwood - Department of Pathology, University of Washington Seattle, Seattle, WA 98195, USA
- Resource Type
- Journal article
- Publication Details
- Experimental cell research, Vol.294(2), pp.406-419
- Publisher
- Elsevier Inc
- DOI
- 10.1016/j.yexcr.2003.11.030
- PMID
- 15023530
- ISSN
- 0014-4827
- eISSN
- 1090-2422
- Language
- English
- Date published
- 2004
- Academic Unit
- Microbiology and Immunology; Radiation Oncology
- Record Identifier
- 9984001151802771
Metrics
17 Record Views