Human fibroblasts expressing hTERT show remarkable karyotype stability even after exposure to ionizing radiation.
Ectopic expression of telomerase results in an immortal phenotype in various types of normal cells, including primary human fibroblasts. In addition to its role in telomere lengthening, telomerase has now been found to have various functions, including the control of DNA repair, chromatin modification, and the control of expression of genes involved in cell cycle regulation. The investigations on the long-term effects of telomerase expression in normal human fibroblast highlighted that these cells show low frequencies of chromosomal aberrations. In this paper, we describe the karyotypic stability of human fibroblasts immortalized by expression of hTERT. The ectopic overexpression of telomerase is associated with unusual spontaneous as well as radiation-induced chromosome stability. In addition, we found that irradiation did not enhance plasmid integration in cells expressing hTERT, as has been reported for other cell types. Long-term studies illustrated that human fibroblasts immortalized by telomerase show an unusual stability for chromosomes and for plasmid integration sites, both with and without exposure to ionizing radiation. These results confirm a role for telomerase in genome stabilisation by a telomere-independent mechanism and point to the possibility for utilizing hTERT-immortalized normal human cells for the study of gene targeting.
Pirzio
L M
LM
CEA-DSV/DRR/LRO, 92265 Fontenay aux roses, France.
Freulet-Marrière
M-A
MA
Bai
Y
Y
Fouladi
B
B
Murnane
J P
JP
Sabatier
L
L
Desmaze
C
C
D002461:Cell Line, Transformed / Q000201:enzymology / Q000528:radiation effects / Q000648:ultrastructure; D002869:Chromosome Aberrations; D002877:Chromosomes, Human / Q000378:metabolism / Q000528:radiation effects*; D002999:Clone Cells / Q000201:enzymology / Q000528:radiation effects / Q000648:ultrastructure; D004268:DNA-Binding Proteins; D005347:Fibroblasts / Q000201:enzymology / Q000528:radiation effects* / Q000648:ultrastructure; D018390:Gene Targeting; D006801:Humans; D007621:Karyotyping; D010957:Plasmids / Q000235:genetics; D011836:Radiation Tolerance; D011993:Recombinant Fusion Proteins / Q000502:physiology; D019098:Telomerase / Q000235:genetics / Q000502:physiology*; D016615:Telomere / Q000648:ultrastructure; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000473:pathology
D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D019098:Telomerase