JOURNAL OF CELLULAR PHYSIOLOGY 173:266–270 (1997)

Telomerase in Human Development

and Cancer
JERRY W. SHAY*
Department of Cell Biology and Neuroscience, The University of Texas Southwestern
Medical Center at Dallas, Dallas, Texas

A potentially rate-limiting step in cancer progression is the conversion of a

normal human cell into one capable of indefinite proliferation. There are at
least two different cellular mechanisms that must be overcome before immor-
talization occurs. The first step generally requires inactivation of the pathways
involving two tumor-suppressor genes, p53 and pRB, and the second step
almost always involves the reactivation of the ribonucleoprotein enzyme te-
lomerase. Telomerase synthesizes hexameric repeats (TTAGGG) onto te-
lomeric ends, thereby compensating for telomeric losses that in its absence
occurs at each cell division. Telomerase is present in human embryonic tissues,
is not detected in most adult tissues, but is upregulated or reactivated in almost
90% of all human cancers. In the present article, I review the telomere –
telomerase theory of aging and cancer including the roles of telomerase during
human development, in differentiation, and in cancer. Research into the regu-
lation of this enzyme may lead to methods to facilitate the accurate diagnosis
of cancer and to the development of novel antitelomerase cancer therapeu-
tics. J. Cell. Physiol. 173:266 – 270, 1997. q 1997 Wiley-Liss, Inc.

TELOMERE – TELOMERASE THEORY OF continued erosion of telomeres that occurs in its ab-
AGING AND CANCER sence during cell division. The enzyme is expressed in
embryonic cells (17) and in adult male germline cells
Although all the steps leading to cancer are as yet (6,17) but is undetectable in normal somatic cells except
unknown, progression to a cancerous state is believed to the proliferative cells of renewal tissues e.g., hemato-
require the accumulation of a series of genetic alterations, poietic stems cell, activated lymphocytes (18 – 21), basal
and an emerging hypothesis is that the upregulation or cells of the epidermis (22 – 23), and intestinal crypt
reexpression of telomerase may be necessary for continu- cells (24). In normal dividing somatic cells, progressive
ous tumor cell growth (1–9). In contrast to normal cells, telomere shortening is observed, eventually leading to
tumor cells show no net loss of average telomere length a limited replicative capacity (25 – 26). It has been pro-
with cell division, suggesting that telomere stability may posed that telomere shortening may be a molecular
be required for cells to escape from replicative senescence measure (clock) of the proliferative potential remaining
and proliferate indefinitely. This notion suggests that in cells (25 – 27). Cells with shortened telomeres stop
most, but not necessarily all, malignant tumors rely on dividing, and this cessation may contribute to altered
acquiring immortality to sustain their growth. Immortali- cellular function as part of aging.
zation may occur through a mutation of a gene in the Thus, the telomere – telomerase hypothesis of aging
telomerase repression pathway, allowing the expression and cancer can be summarized as follows: (a) progres-
of telomerase in cancer cells. Thus, it is becoming more sive telomere loss in dividing cells is normal and may
widely accepted that the upregulation (7) or reactivation be the ‘‘clock’’ or timing mechanism that regulates
(8) of telomerase activity may be a rate-limiting if not cellular senescence; (b) cellular senescence occurs when
the critical step required for the continuing proliferation telomeres are short, although the molecular mecha-
of advanced cancers. nism for this phenomenon is not clear; (c) mutation(s)
Telomeres are thought to protect the ends of all linear
chromosomes, allow their complete replication, and
help maintain genetic stability (10 – 12). In the absence Contract grant sponsor: National Institutes of Health; Contract
grant number: AG07992; Contract grant sponsor: Geron Corpora-
of a mechanism to maintain telomeres, progressive tion, Menlo Park, California; Contract grant sponsor: Texas Ad-
shortening occurs with each cell division due to the vanced Technology Program; Contract grant number: 003660-
end-replication problem (13 – 14), eventually leading to 041; Contract grant number: USAMR; Contract grant number:
cessation of cell proliferation. Telomerase is a ribo- DAMD17-94-J-4077.
nucleoprotein enzyme that stabilizes telomere length *Correspondence to: J.W. Shay, Department of Cell Biology and
by adding hexameric (TTAGGG) repeats (15 – 16) to the Neuroscience, The University of Texas Southwestern Medical
telomeric ends of chromosomes, compensating for the Center at Dallas, 5323 Harry Hines Boulevard, Dallas, TX 75235.
q 1997 WILEY-LISS, INC.

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 TELOMERASE IN HUMAN DEVELOPMENT AND CANCER 267

occur in the telomerase repression pathway, which re- (37,50 – 51). Except for differentiated cells, which are
activates or upregulates the enzyme; (d) telomerase ac- postmitotic, this repression during G0 is fully revers-
tivity correlates with the stabilization of telomere ible. Most somatic cells are telomerase silent and lack
length and cellular immortalization; and (e) telomerase telomerase activity both when quiescent and when di-
activity is necessary for the continued proliferation of viding, whereas telomerase-competent cells, although
cells and is a critical, perhaps rate-limiting, step in telomerase negative when quiescent, can express te-
cancer progression. lomerase activity upon G0 exit. Hence, telomerase is
likely repressed by a pathway different from that which
TELOMERASE ACTIVITY ASSAY represses telomerase in proliferating normal somatic
A highly sensitive assay, based on polymerase chain cells.
reaction (PCR), for measuring telomerase activity has These observations lead to two important concepts
been developed that includes an improved detergent regarding the level of telomerase activity in a popula-
lysis method to allow more uniform extraction of te- tion of telomerase-competent cells. If most cells in a
lomerase from a small number of cells (6). In the te- mixed population are in G0, telomerase activity will be
lomeric repeat amplification protocol (TRAP) assay, te- predicted to be very low, whereas if most are dividing
lomerase first synthesizes extension products, which the activity will be much higher. Because telomerase
then serve as templates for PCR amplification. The in- activity does not change during G1, S, and G2 / M,
creased sensitivity of this assay has resulted in a dra- the activity in the population of cells may not reflect
matic increased number of investigations (28 – 43). the rate of cell division but rather the fraction of cells
More recent improvements in the assay include the use that are cycling. A second important concept is that
of an internal standard to permit semiquantitation of because telomerase-competent cells can regulate the
activity and detection of amplification inhibitors pres- level of telomerase activity, the presence of telomerase
ent in some tissue extracts (44). Nonradioactive does not necessarily imply stable telomere lengths. It
methods for detecting telomerase have been reported has been documented that although cells of renewal
(20), and the recent introduction of a research kit tissues such as blood (18 – 22,52), skin (22 – 23), and
(TRAPezey, Oncor, Gaithersburg, MD) for detecting intestinal epithelia (24) have detectable telomerase ac-
telomerase activity makes possible a standardized tivity, the levels are insufficient to prevent telomere
assay that may improve reproducibility among labora- shortening. The most primitive ‘‘stem’’ cells of such re-
tories (45). newal tissues are likely to be quiescent much of the
time and exhibit a low level of telomerase activity.
TELOMERASE ACTIVITY DURING However, with stimulation to proliferate, telomerase
DEVELOPMENT activity appears to be upregulated in their immediate
Initial studies have indicated the presence of te- progeny, most probably to help slow the rate of telomere
lomerase activity in human testis and fetal ovary and erosion during rapid turnover of such cells. However,
adrenal glands (6). Recently, we reported that although the more mature progeny revert to quiescence downreg-
telomerase activity cannot be detected in human ma- ulating telomerase activity.
ture sperm or unfertilized eggs, it is detected as early
as the blastocyst stage of embryogenesis (17). Although TELOMERASE ACTIVITY IN THE EARLY
its activity is found in a wide variety of tissues at 16 DETECTION OF CANCER
weeks gestation, it is not present in fetal brain at that Telomerase activity can be detected in most primary
time, nor is it detected in liver, skin, lung, muscle, or human tumor specimens and tumor-derived cell lines.
adrenal tissue following birth or in most adult tissues. Its activity is detected in 85 – 95% of the most common
The lack of activity in human oocytes and mature cancers such as prostate (28), breast (36,40), colon
human sperm contrasts with reports of telomerase ac- (24,30,41,42), lung (34), and liver (29) and in some in-
tivity in Xenopus eggs and sperm (46). Because sperm stances at an in situ stage. In breast and lung cancers,
telomeres do not shorten with age (47 – 48) and W/Wv telomerase activity appears very early (even in preneo-
mice that lack primary spermatogonia also lack testicu- plasia of smokers and former smokers); in pancreatic
lar telomerase activity (49), the telomerase activity in (Hiyama et al., personal communication) and colon can-
testis extracts probably is derived from germline cells cers, it appears in 90 – 95% of early stage carcinoma
at the spermatogonial or spermatocyte stage. Recent but not in preneoplasia (24). There is evidence in neuro-
studies examining the presence of telomerase RNA in blastoma (33), acute myeloid leukemia (31,37), breast
seminiferous tubules of adult human testis by in situ (36,40), and gastrointestinal (30,35) cancers that the
analysis confirm that at least the RNA component of presence of high levels of telomerase correlates with
telomerase is detected in spermatogonial cells and poor prognosis.
some spermatocytes (unpublished observations). Almost any type of clinical specimen can be utilized
to detect telomerase activity including exfoliated cells
TELOMERASE IN QUIESCENCE AND and fine-needle aspirates, and several applications
DIFFERENTIATION have been described (6,24,34,36,40). Telomerase activ-
In telomerase-competent proliferating cells, activity ity may even be detected in cells sedimented from urine
is detected in the G1, S, and G2 / M phases of the cell in bladder cancer (D. Tarin and M. Muller, personal
cycle (46,50) but is repressed when such cells enter G0 communication). The use of minimally invasive proce-
due to growth factor deprivation (50), contact inhibition dures such as fine-needle aspirations (36,40) in breast
of cell division (50), the induction of senescence in re- cancer holds great promise for aiding in clinical diagno-
versibly immortalized cells (50), or differentiation sis prior to surgery and in some instances in which

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268 SHAY

the cytology is not informative may help distinguish ist (54 – 55) in cell culture models of immortalization,
carcinoma from benign disease. Telomerase analysis of their roles in vivo are undefined. Although there is hope
cytological samples obtained by flexible fiberoptic bron- that studying aspects of telomerase in animal models
choscopy and by thoracocentesis, procedures that aid of carcinogenesis may yield insights into basic mecha-
in identifying lung carcinoma, may also aid the patholo- nisms of telomere and telomerase regulation, it is im-
gist (K. Hiyama personal communication). Also, te- portant that we recognize that such regulation may
lomerase levels in oral rinses (D. Sidransky, personal differ between species, reflected in the wide difference
communication) may predict premalignant leukoplakia of cancer susceptibility of mice versus humans.
lesions capable of progressing to carcinoma (43). Al- Telomere length measurements are unlikely to be
though analysis of colonic effluents and washes (M. of clinical usefulness in tumors that have telomerase
Tobi and D. Tarin, personal communication) may detect activity but may be clinically relevant when telomerase
early-stage colorectal carcinoma, more studies are re- activity is not elevated (39). Importantly, we have
quired for confirmation. Telomerase activity may be a learned that the detection of telomerase activity only
marker of minimal residual disease in patients under- indicates the potential for cellular immortality and is
going chemotherapy, and its detection also may be a not always synonymous with cancer development (9).
useful surrogate for following efficacy of chemopreven- In some cases of metastatic neuroblastoma, tumors are
tion trials. In a recent study (53), although fewer than observed that eventually undergo remission (33); there-
20% of ordinary meningiomas were shown to have te- fore, ‘‘mortal’’ tumors are not incompatible with the
lomerase activity, those with detectable activity had telomere – telomerase theory of aging and cancer. Inter-
a very high probability of tumor recurrence. Finally, pretation of telomerase activity in tissue extracts can
brushings to analyze telomerase activity in pancreatic be complicated by quiescence of telomerase-competent
secretions (E. Hiyama, personal communication) and cells, telomerase expression by inflammatory cells, and
fine-needle biopsies for distinguishing thyroid adeno- by the presence of telomerase assay inhibitors (9).
mas from carcinomas (S. Sukumar, personal communi- Thus, repression of telomerase activity in quiescence
cation) appear to be promising ways to distinguish pre- suggests that different levels of telomerase in tumor
neoplastic from carcinoma tissues. biopsies may reflect the fraction of proliferating cells
Preliminary data indicate that during the progres- (9,50), indicating the need for the development of a
sion of lung cancer the detection of telomerase activity telomerase assays for tissue samples in situ. The recent
appears very early. Multiple specimens of lung and cloning of the RNA component of telomerase (56) makes
bronchial epithelium at different distances from the in situ analysis of archival tissue specimens feasible.
invasive tumor from 10 patients undergoing surgery Such data may provide important insights into the reg-
were examined (A. Gazdar, unpublished observations), ulation of the enzyme.
as were samples of carcinoma in situ obtained by fluo- In addition, telomerase inhibitors are being actively
rescent bronchoscopy. Although most of these cases considered as potential antitumor agents, with the ex-
were from smokers or former smokers, in such samples pectation that lack of telomerase expression in normal
progressively higher percentage of samples with hyper- somatic cells will result in a highly specific treatment
plasia, metaplasia, and dysplasia expressed activity as with fewer side effects than occurs with conventional
did all five samples of carcinoma in situ, although 15% chemotherapy (57). Future strategies may include con-
of histologically normal epithelium expressed te- ventional chemotherapies to reduce tumor mass fol-
lomerase. The relative amounts of telomerase activity lowed by a recovery period to permit renewed prolifera-
in preneoplastic specimens and in the carcinoma in tion of primitive stem cells to replace damaged tissues.
situ, after adjustment for the number of epithelial cells With their return to quiescence, antitelomerase treat-
in the specimen, were approximately 10-fold lower than ments perhaps in combination with other novel cancer
in the corresponding tumor. Thus, although there are targets such as anti-angiogenic agents could limit the
low levels of telomerase activity early during pathogen- proliferative capacity of surviving tumor cells.
esis of lung cancer, an upregulation occurs at the time
of invasive cancer. In situ studies examining for the SUMMARY
presence of telomerase RNA confirm that telomerase Although there is justifiable optimism regarding te-
RNA is focally present in preinvasive lung lesions of lomerase activity as a useful tool in the early detection
smokers and former smokers (A. Gazdar, unpublished of cancer and possible as a prognostic indicator of out-
observations). Such studies should clarify the nature come, additional validation studies are required before
of the changes in telomerase activity during the patho- knowledge of telomerase activity will be useful in a
genesis of this disease. practical sense for decisions regarding patient manage-
ment. A key question is whether telomerase detection
FUTURE DIRECTIONS will allow distinction between quiescent and progress-
Many aspects of the complex biology and basic regu- ing cancers. Molecular staging using markers such as
lation of telomerase remain unresolved. Additional telomerase activity in combination with other molecu-
studies of pathways involved in the regulation of lar markers may hold great promise in this regard.
telomere length and telomerase activity are being ac- Therefore, knowledge of telomerase in clinical speci-
tively pursued. These pursuits would include more mens may help risk stratify individual patients to de-
accurate methods for measuring telomere lengths, bet- termine which treatments may be beneficial. Finally,
ter telomerase extraction methods, and identification of the results of clinical trials regarding the utility of in-
genes regulating the repression pathway of telomerase. hibitors of telomerase in the therapy of cancer are
Although alternate pathways to immortality clearly ex- awaited with great anticipation.

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 TELOMERASE IN HUMAN DEVELOPMENT AND CANCER 269

LITERATURE CITED 26. Harley, C.B. (1995) Telomeres and aging: Fact, fancy, and the
future. J. NIH Res., 7:64 – 68.
27. Allsopp, R.C., Vaziri, H., Patterson, C., Goldstein, S., Younglai,
1. Bacchetti, S., and Counter, C.M. (1995) Telomeres and telomerase E.V., Futcher, A.B., Greider, C.W., and Harley, C.B. (1992)
in human cancer. Int. J. Oncol., 7:423 – 432. Telomere length predicts replicative capacity of human fibro-
2. Shay, J.W., and Wright, W.E. (1996) Telomerase activity in hu- blasts. Proc. Natl. Acad. Sci. USA, 89:10114 – 10118.
man cancer. Curr. Opin. Oncol., 8:66 – 71. 28. Sommerfeld, H.J., Meeker, A.K., Piatyszek, M.A., Bova, G.S.,
3. de Lange, T. (1994) Activation of telomerase in a human tumor. Shay, J.W., and Coffey, D.S. (1996) Telomerase activity distin-
Proc. Natl. Acad. Sci. USA., 91:2882 – 2885. guishes benign from malignant human prostate tissue. Cancer
4. Counter, C.M., Avilion, A.A., LeFeuvre, C.E., Stewart, N.G., Res., 56:218 – 222.
Greider, C.W., Harley, C.B., and Bacchetti, S. (1992) Telomere 29. Tahara, H., Nakanishi, T., Kitamoto, M., Nakashio, R., Shay,
shortening associated with chromosome instability is arrested in J.W., Tahara, E., Kajiyama, G., and Ide., T. (1995) Telomerase
immortal cells which express telomerase activity. EMBO J., activity in human liver tissues: Correlation between chronic liver
11:1921 – 1929. disease and hepatocellular carcinoma. Cancer Res., 55:2734 –
5. Counter, C.M., Hirte, H.W., Bacchetti, S., and Harley, C.B. (1994) 2736.
Telomerase activity in human ovarian carcinoma. Proc. Natl. 30. Tahara, H., Kuniyasu, H., Yasui, W., Yokozaki, H., Shay, J.W.,
Acad. Sci. USA, 91:2900 – 2904. Ide, T., and Tahara, E. (1995) Telomerase activity in gastric and
6. Kim, N.W., Piatyszek, M.A., Prowse, K.R., Harley, C.B., West, colorectal carcinomas correlates well with malignant progression.
M.D., Ho, P.L.C., Coviello G.M., Wright W.E., Weinrich S.L., and Clin. Cancer Res. Rapid Commun., 1:1245 – 1251.
Shay, J.W. (1994) Specific association of human telomerase activ- 31. Counter, C.M., Gupta, J., Harley, C.B., Leber, B., and Bacchetti,
ity with immortal cells and cancer. Science, 266:2011 – 2015. S. (1995) Telomerase activity in normal leukocytes and in hemato-
7. Greaves, M. (1996) Telomerase activation, stem cells and cancer. logic malignancies. Blood, 85:2315 – 2320.
Trends Genet., 12:127 – 128. 32. Langford, L.A., Piatyszek, M.A., Xu, R., Schold, S.C., and Shay,
8. Shay, J.W., and Wright, W.E. (1996) The reactivation of te- JW. (1995) Telomerase activity in human brain tumors. Lancet,
lomerase activity in cancer progression. Trends Genet., 12:129 – 346:267 – 1268.
131. 33. Hiyama, E., Hiyama, K., Yokoyama, T., Matsuura, Y., Piatyszek,
9. Holt, S.E., Shay, J.W., and Wright, W.E. (1996) Refining the M.A., and Shay, J.W. (1995) Correlating telomerase activity levels
telomere-telomerase hypothesis of aging and cancer. Nature Bio- with human neuroblastoma outcomes. Nature Med., 1:249 – 255.
tech., 14:836 – 840. 34. Hiyama, K., Hiyama E., Ishioka S., Yamakido M., Inai K., Gazdar
10. Blackburn, E.H. (1991) Structure and function of telomeres. Na- A.F., Piatyszek M.A., and Shay J.W. (1995) Telomerase activity in
ture, 350:569 – 573. small-cell and non-small-cell lung cancers. J. Natl. Cancer Inst.,
11. Blackburn, E.H. (1994) Telomeres: No end in sight. Cell, 77:621 – 87:895 – 902.
623. 35. Hiyama, E., Yokoyama T., Tatsumoto N., Hiyama K., Imamura
12. Greider, C. (1994) Mammalian telomere dynamics: Healing, frag- Y., Murakami Y. Kodama T., Piatyszek M.A., Shay J.W., and
mentation shortening and stabilization. Curr. Opin. Genet. Dev., Matsuura Y. (1995) Telomerase activity in gastric cancer. Cancer
4:203 – 211. Res., 55:3258 – 3262.
13. Olovnikov, A.M. (1971) Principles of marginotomy in template 36. Hiyama, E., Hiyama, E., Ishioka, S., Yamakido, M., Inai, K.,
synthesis of polynucleotides. Doklady Akad. Nauk. SSSR, Gazdar, A.F., Piatyszek, M.A., and Shay, J.W. (1996) Telomerase
201:1496 – 1499. activity in human breast tumors. J. Natl. Cancer Inst., 88:116 –
14. Watson, J.D. (1972) Origin of concatameric T7 DNA. Nature New 122.
Biol. (Lond.), 239:197 – 201. 37. Zhang, W., Piatyszek, M.A., Estey, E., Kobayashi, T., Andreff, M.,
15. Morin, G.B. (1989) The human telomerase terminal transferase Deisseroth, A.B., and Shay, J.W. (1996) Telomerase activity in
enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. acute myelogenous leukemia: Inhibition of telomerase activity by
Cell, 59:521 – 529. differentiation-inducing agent. Clin. Cancer Res. Adv. Brief,
16. Moyzis, R.K., Buckingham, J.M., Cram, L.S., Dani, M., Deaven, 2:799 – 803.
L.L., Jones, M.D., Meyne, J., Ratliff, R.L., and Wu, J.R. (1988) A 38. Mehle, C., Piatyszek, M.A., Ljungberg, B., Shay, J.W., and Roos,
highly conserved repetitive DNA sequence, (TTAGGG)n , present G. (1996) Telomerase activity in human renal cell carcinoma. On-
at the telomeres of human chromosomes. Proc. Natl. Acad. Sci. cogene, 13:161 – 166.
USA, 85:6622 – 6626. 39. Shay, J.W., Werbin, H., and Wright, W.E. (1996) Telomeres and
17. Wright, W.E., Piatyszek, M.A., Rainey, W.E., Byrd, W., and Shay, telomerase in human leukemias. Leukemia, 10:1255 – 1261.
J.W. (1996) Telomerase activity in human germline and embry- 40. Sugino, T., Yoshida, K., Bolodeoku, J., Tahara, H., Buley, I., Ma-
onic tissues and cells. Dev. Genet., 18:173 – 179. nek, S., and Tarin, D. (1996) Telomerase activity in human breast
18. Hiyama, K., Hirai Y., Kyoizumi S., Akiyama M., Hiyama E., Pia- cancer and benign breast lesions: Diagnostic applications in clini-
tyszek M.A., Shay J.W., Ishioka S., and Yamakido M. (1995) Acti- cal specimens including fine needle aspirates. Int. J. Cancer
vation of telomerase in human lymphocytes and hematopoietic 69:301 – 306.
progenitor cells. J. Immunol., 155:3711 – 3715. 41. Chadeneau, C., Hay, K., Hirte, H.W., Galligner, S., and Bacchetti,
19. Broccoli, D., Young, J.W., and de Lange, T. (1995) Telomerase S. (1995) Telomerase activity associated with acquisition of malig-
activity in normal and malignant hematopoietic cells. Proc. Natl. nancy in human colorectal cancer. Cancer Res., 55:2533 – 2536.
Acad. Sci. USA, 92:9082 – 9086. 42. Li, Z.-H., Salovaara, R., Aaltonen, L.A., and Shibata, D. (1996)
20. Piatyszek, M.A., Kim N.W., Weinrich S.L., Hiyama, K., Hiyama, Telomerase activity is commonly detected in hereditary nonpolyp-
E., Wright, W.E., and Shay, J.W. (1995) Detection of telomerase osis colorectal cancer. Am. J. Pathol., 148:1075 – 1079.
activity in human cells and tumors by a telomeric repeat amplifi- 43. Mutirangura, A., Supiyaphun, P., Trirekapan, S., Sriuranpong,
cation protocol (TRAP). Methods Cell Sci., 17:1 – 15. V., Sakuntabhai, A., Yenrudi, S., and Voravud, N. (1996) Te-
21. Chiu, C.P., Dragowska, W., Kim, N.W., Vaziri, H., Yui, J., lomerase activity in oral leukoplakia and head and neck squa-
Thomas, T.E., Harley, C.B., and Lansdorp, P.M. (1996) Differen- mous cell carcinoma. Cancer Res., 56:3530 – 3533.
tial expression of telomerase activity in hematopoietic progenitors 44. Wright, W.E., Shay, J.W., and Piatyszek, M.A. (1995) Modifica-
from adult human bone marrow. Stem Cells, 14:239 – 248. tion of a telomeric repeat amplification protocol (TRAP) results
22. Taylor, R.S., Ramirez, R.D., Ogoshi, M., Chaffins, M., Piatyzsek, in increased reliability, linearity and sensitivity. Nucleic Acids
M.A., and Shay, J.W. (1996) Telomerase activity in malignant Res., 23:3794 – 3795.
and nonmalignant skin conditions. J. Invest. Dermatol., 106:759 – 45. Holt, S.E., Norton, J.C., Wright W.E., and Shay, J.W. (1996) Com-
765. parison of the telomeric repeat amplification protocol (TRAP) to
23. Harle-Bachor, C., and Boukamp, P. (1996) Telomerase activity in the new TRAP-ezeTM telomerase detection kit. Methods Cell Sci.
the regenerative basal layer of the epidermis in human skin and 18:237 – 248.
in immortal and carcinoma-derived skin keratinocytes. Proc. 46. Mantell, L.L., and Greider, C.W. (1994) Telomerase activity in
Natl. Acad. Sci. USA, 93:6476 – 6481. germline and embryonic cells of Xenopus. EMBO J., 13:3211 –
24. Hiyama, E., Hiyama, K., Tatsumoto, N., Shay, J.W., and Yoko- 3217.
yama, T. (1996) Telomerase activity in human intestine. Int. J. 47. Harley, C.B., Futcher, B.A., and Greider, C.W. (1990) Telomeres
Oncol., 9:453 – 458. shorten during ageing of human fibroblasts. Nature, 345:458 –
25. Harley, C.B. (1991) Telomere loss: Mitotic clock or genetic time 460.
bomb? Mutat. Res., 256:271 – 282. 48. Vaziri, H., Dragowska, W., Allsopp, R.C., Thomas, T.E., Harley,

/ 8928$$550d 09-24-97 15:14:17 wlcpas W Liss: JCP JCP-550D

270 SHAY

C.B., and Landsdorp, P.M. (1994) Evidence for a mitotic clock in 53. Langford, L.A., Piatyszek, M.A., Xu, R., Schold, S.C., Wright,
human hematopoietic stem cells: Loss of telomeric DNA with age. W.E., and Shay, J.W. (1997) Telomerase activity: A prognostic
Proc. Natl. Acad. Sci. USA, 91:9857 – 9860. indicator in ordinary meningiomas Hum. Pathol. 28:416 – 420.
49. Prowse, K.R., and Greider, C.W. (1995) Developmental and tissue 54. Murnane, J.P., Sabatier, L., Marder, B.A., and Morgan, W.F.
specific regulation of mouse telomerase and telomere length. Proc. (1994) Telomere dynamics in an immortal human cell line. EMBO
Natl. Acad. Sci. USA, 92:4818 – 4822. J., 13:4953 – 4962.
50. Holt, S.E., Wright, W.E., and Shay, J.W. (1996) Regulation of 55. Bryan, T.M., Englezou, A., Gupta, J., Bacchetti, S., and Reddel,
telomerase activity in immortal cell lines. Mol. Cell. Biol., R.R. (1995) Telomere elongation in immortal human cells without
16:2932 – 2939. detectable telomerase activity. EMBO J., 14:4240 – 4248.
51. Sharma, H.W., Sokoloski, J.A., Perez, J.R., Maltese, J.Y., Sart-
orelli, A.C., Stein, C.A., Nichols, G., Khaled, Z., Telang, N.T., 56. Feng, J., Funk, W.D., Wang, S.-S., Weinrich, S.L., Avilion, A.A.,
and Narayanan, R. (1995) Differentiation of immortal cells in- Chiu, C.-P., Adams, R.R., Chang E., Allsopp R.C., Yu J., Le S.,
hibits telomerase activity. Proc. Natl. Acad. Sci. USA, West M.D., Harley C.B., Andrews W.H., Greider C.W., and Ville-
92:12343 – 12346. ponteau B. (1995) The RNA component of human telomerase.
52. Weng, N.-P., Levine, B.L., June, C.H., and Hodes, R.J. (1995) Science, 269:1236 – 1241.
Human naive and memory T lymphocytes differ in telomeric 57. Norton, J.C., Piatyszek, M.A., Wright, W.E., Shay, J.W., and
length and replicative potential. Proc. Natl. Acad. Sci. USA, Corey, D.R. (1996) Inhibition of human telomerase activity by
92:11091 – 11094. peptide nucleic acid oligomers. Nature Biotech., 14:615 – 619.

/ 8928$$550d 09-24-97 15:14:17 wlcpas W Liss: JCP JCP-550D