Transient expression of human telomerase extends the life span of normal human fibroblasts

被引:110
作者
Steinert, S [1 ]
Shay, JW [1 ]
Wright, WE [1 ]
机构
[1] Univ Texas, SW Med Ctr, Dept Cell Biol, Dallas, TX 75390 USA
关键词
telomerase; telomeres; Cre/lox-recombination;
D O I
10.1006/bbrc.2000.3080
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We utilized the Cre/lox recombination system to transiently express the catalytic subunit of telomerase (hTERT) in normal diploid foreskin fibroblasts (BJ cells). A retroviral construct containing an hTERT cDNA, flanked by loxP-sites was introduced into near senescent BJ cells (population doubling 85). At population doubling (PD) 92, which exceeds the typical life span of these cells, we excised the gene via Cre-mediated recombination. All clones lost telomerase activity and showed telomere shortening over an additional 50 PDs. Interestingly, the average telomere length in these cells became shorter than in untreated BJ cells at senescence, This may be due to hTERT preferentially elongating the shortest telomeres, leading to greater length uniformity, In summary, transient telomerase expression and only a very small average telomere elongation by hTERT resulted in a 50% increase in life span of human fibroblasts. This suggests a potentially safe use of hTERT in tissue engineering. (C) 2000 Academic Press.
引用
收藏
页码:1095 / 1098
页数:4
相关论文
共 21 条
[1]   EVIDENCE FOR A CRITICAL TELOMERE LENGTH IN SENESCENT HUMAN FIBROBLASTS [J].
ALLSOPP, RC ;
HARLEY, CB .
EXPERIMENTAL CELL RESEARCH, 1995, 219 (01) :130-136
[2]   EXCISION OF SPECIFIC DNA-SEQUENCES FROM INTEGRATED RETROVIRAL VECTORS VIA SITE-SPECIFIC RECOMBINATION [J].
BERGEMANN, J ;
KUHLCKE, K ;
FEHSE, B ;
WOLFRAM, IR ;
LOTHER, H .
NUCLEIC ACIDS RESEARCH, 1995, 23 (21) :4451-4456
[3]   Extension of life-span by introduction of telomerase into normal human cells [J].
Bodnar, AG ;
Ouellette, M ;
Frolkis, M ;
Holt, SE ;
Chiu, CP ;
Morin, GB ;
Harley, CB ;
Shay, JW ;
Lichtsteiner, S ;
Wright, WE .
SCIENCE, 1998, 279 (5349) :349-352
[4]   A BIOMARKER THAT IDENTIFIES SENESCENT HUMAN-CELLS IN CULTURE AND IN AGING SKIN IN-VIVO [J].
DIMRI, GP ;
LEE, XH ;
BASILE, G ;
ACOSTA, M ;
SCOTT, C ;
ROSKELLEY, C ;
MEDRANO, EE ;
LINSKENS, M ;
RUBELJ, I ;
PEREIRASMITH, O ;
PEACOCKE, M ;
CAMPISI, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (20) :9363-9367
[5]   TELOMERE LOSS - MITOTIC CLOCK OR GENETIC TIME BOMB [J].
HARLEY, CB .
MUTATION RESEARCH, 1991, 256 (2-6) :271-282
[6]   LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS [J].
HAYFLICK, L .
EXPERIMENTAL CELL RESEARCH, 1965, 37 (03) :614-&
[7]   SERIAL CULTIVATION OF HUMAN DIPLOID CELL STRAINS [J].
HAYFLICK, L ;
MOORHEAD, PS .
EXPERIMENTAL CELL RESEARCH, 1961, 25 (03) :585-+
[8]  
Holt Shawn E., 1996, Methods in Cell Science, V18, P237, DOI 10.1007/BF00132889
[9]   Transient expression of SV 40 large T antigen by Cre/LoxP-mediated site-specific deletion in primary human tumor cells [J].
Li, LP ;
Schlag, PM ;
Blankenstein, T .
HUMAN GENE THERAPY, 1997, 8 (14) :1695-1700
[10]   A SERIES OF MAMMALIAN EXPRESSION VECTORS AND CHARACTERIZATION OF THEIR EXPRESSION OF A REPORTER GENE IN STABLY AND TRANSIENTLY TRANSFECTED CELLS [J].
MORGENSTERN, JP ;
LAND, H .
NUCLEIC ACIDS RESEARCH, 1990, 18 (04) :1068-1068