Telomerase activity during spontaneous immortalization of Li-Fraumeni syndrome skin fibroblasts.
Li-Fraumeni Syndrome (LFS) is characterized by heterozygous germline mutations in the p53 gene. Accompanied by genomic instability and loss or mutation of the remaining wild type p53 allele, a low frequency of spontaneous immortalization in LFS fibroblasts occurs. It is believed that the loss of p53 wild type function contributes to immortalization of these LFS fibroblasts, but it is not clear if this is sufficient. Because stabilization of telomere length is also thought to be a necessary step in immortalization, telomerase activity, expression of the telomerase RNA component (hTR) and telomere length were anlaysed at various passages during the spontaneous immortalization of LFS skin fibroblasts. One LFS strain which immortalized, MDAH087 (087), had no detectable telomerase activity whereas another LFS strain which immortalized, MDAH041 (041), had detectable telomerase activity. In preimmortal cells from both strains, hTR was not detected by in situ hybridization. Immortal 087 cells remained negative for hTR, while immortal 041 cells demonstrated strong hTR in situ hybridization signals. 087 cells had long and heterogenous telomeres whereas telomeres of 041 cells had short, stable telomere lengths. Tumorigenicity studies in nude mice with ras-transformed 087 and 041 cells resulted in both cell lines giving rise to tumors and retaining telomerase status. Overall these results suggest that strain specificity may be important in telomerase re-activation and that both abrogation of p53 function and a mechanism to maintain telomeres are necessary for immortalization.
Department of Cell Biology and Neuroscience, UT Southwestern Medical Center, Dallas 75235-9039, USA.
L S
LS
Gollahon
E
E
Kraus
T A
TA
Wu
S O
SO
Yim
L C
LC
Strong
J W
JW
Shay
M A
MA
Tainsky
D000818:Animals; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D016922:Cellular Senescence; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D006801:Humans; D016864:Li-Fraumeni Syndrome / Q000201:enzymology*; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D009374:Neoplasms, Experimental; D012313:RNA / Q000032:analysis*; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D012867:Skin / Q000166:cytology / Q000201:enzymology*; D019098:Telomerase / Q000032:analysis*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D016159:Tumor Suppressor Protein p53; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase