Immortalization in a normal foreskin fibroblast culture following transduction of cyclin A2 or cdk1 genes in retroviral vectors
| Autor: | Yoshihiro Takihara, Xiaoqin Chen, Thomas H. Norwood, Motoaki Ohtsubo, Matthew D. Gray, Maria Tresini, Anthony C. Saulewicz, Deborah E. Banker, Ping Luo, Aloysius L. Klingelhutz, Vincent J. Cristofalo |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Cyclin-Dependent Kinase Inhibitor p21
Male Telomerase Population Genetic Vectors Loss of Heterozygosity Cyclin A Biology Retinoblastoma Protein Viral vector Transduction Genetic Cyclins CDC2 Protein Kinase Humans education Cellular Senescence Cyclin-Dependent Kinase Inhibitor p16 Cell Line Transformed Chromosome Aberrations education.field_of_study Cyclin-dependent kinase 1 Infant Newborn Cell Biology Fibroblasts Molecular biology Telomere Clone Cells Retroviridae Cell culture Tumor Suppressor Protein p53 Immortalised cell line Cyclin A2 Cell Division |
| Zdroj: | Experimental cell research. 294(2) |
| ISSN: | 0014-4827 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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