Abstract
Telomerase activity, the ability to add telomeric repeats to the ends of chromosomes, has been detected in most immortal cell lines including tumor cells, but is low or absent in most diploid, mortal cells such as those of somatic tissues. Peptide nucleic acids (PNAs), analogs of DNA or RNA which bind to complementary nucleic acids with very high affinity, were co-electroporated into immortal human cells along with a selectable plasmid. Introduction of PNAs inverse-complementary to telomerase RNA effectively inhibited telomerase activity in intact cells, shortened telomeres, reduced colony size, and arrested cell proliferation after a lag period of 5 – 30 cell generations, consistent with suppression of their `immortality'. Electroporation of selection plasmid alone had no effect, while PNAs of altered sequence were markedly less effective in each assay. This constitutes the first demonstration of cell growth arrest through telomerase inhibition, upon treatment of intact cells with an exogenous compound which can be efficiently delivered in vivo. The phenotype of telomerase-inhibited transformed cells differs from senescence of normal diploid fibroblasts, but rather resembles the crisis state of incompletely transformed cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Banga SS, Kim S-H, Hubbard K, Dasgupta T, Jha KK, Patsalis P, Hauptschein R, Gamberi B, Dalla-Favera R, Kraemer P and Ozer HL. . 1997 Oncogene 14: 313–321.
Blackburn EH. . 1992 Annu. Rev. Biochem. 61: 113–129.
Blasco MA, Lee H-W, Hande MP, Samper E, Lansdorp PM, DePinho RA and Greider CW. . 1997 Cell 91: 25–34.
Bodnar AG, Ouellette M, Frolkis M, Holt SE, Chiu C-P, Morin GB, Harley CB, Shay JW, Lichsteiner S and Wright WE. . 1998 Science 279: 349–352.
Broccoli D, Young JW and de Lange T. . 1995 Proc. Natl. Acad. Sci. USA 92: 9082–9086.
Cachianes G, Ho C, Weber RF, Williams SR, Goeddel DV, Leung DW. . 1993 Biotechniques 15: 255–259.
Cheng RZ, Shammas MA, Li J and Shmookler Reis RJ. . 1997 Exp. Cell Res. 234: 300–312.
Chu G, Hayakawa H and Berg P. . 1987 Nucleic Acids Res. 15: 1311–1326.
Counter CM, Avilion AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB and Bacchetti S. . 1992 EMBO J. 11: 1921–1929.
Cristofalo VJ and Sharf BB. . 1973 Exp. Cell Res. 76: 419–427.
Demidov VV, Potaman VN, Frank-Kamenetskii MD, Egholm M, Buchard O, Sonnichsen SH and Nielsen PE. . 1994 Biochem. Pharmacol. 48: 1310–1313.
Egholm M, Buchardt O, Christensen L, Behrens C, Freier SM, Driver DA, Berg RH, Kim SK, Norden B and Nielsen PE. . 1993 Nature 365: 566–568.
Fedoroff OY, Salazar M, Han H, Chemeris VV, Kerwin SM and Hurley LH. . 1998 Biochemistry 37: 12367–12374.
Feng J, Funk WD, Wang S-S, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, Le S, West MD, Harley CB, Andrews WH, Greider CW and Villeponteau B. . 1995 Science 269: 1236–1241.
Fletcher TM, Salazar M and Chen S-F. . 1996 Biochemistry 35: 15611–15617.
Hamilton SE, Pitts AE, Katipally RR, Jia X, Rutter JP, Davies BA, Shay JW, Wright WE and Corey DR. . 1997 Biochemistry 36: 11873–11880.
Harley CB and Goldstein S. . 1978 J. Cell. Physiol. 97: 509–516.
Harley CB, Futcher BA and Greider CW. . 1990 Nature 345: 458–460.
Hayflick L. . (1977) Handbool of the Biology of Aging. Finch CE and Hayflick L (eds.). Van Nostrand Reinhold Co.: NY 159–186.
Holt SE, Norton JC, Wright WE and Shay JW. . 1996 Meth. Cell Sci. 18: 237–248.
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL and Shay JW. . 1994 Science 266: 2011–21015.
Kondo S, Kondo Y, Li G, Silverman RH and Cowell JK. . 1998a Oncogene 16: 3323–3330.
Kondo S, Tanaka Y, Kondo Y, Hitomi M, Barnett GH, Ishizaka Y, Liu J, Haqqi T, Nishiyama A, Villeponteau B, Cowell JK and Barna BP. . 1998b FASEB J. 12: 801–811.
Lee H-W, Blasco MA, Gottlieb GJ, Horner JW II, Greider CW and DePinho RA. . 1998 Nature 392: 569–574.
Martin GM, Sprague C and Epstein C. . 1970 Lab. Invest. 23: 86–91.
Moerman EJ and Goldstein S. . (1986) Methods in Diabetes Research, Vol. II, Clinical Methods. Clarke WL, et al. (eds.). John Wiley & Sons: NY 283–312.
Nielson PE, Egholm M, Berg RH and Buchardt O. . 1991 Science 254: 1497–1500.
Norton JC, Piatyszek MA, Wright WE, Shay JW and Corey DR. . 1996 Nature Biotech. 14: 615–619.
Norton JC, Waggenspack JH, Varnum E and Corey DR. . 1995 Bioorg. Med. Chem. 3: 437–445.
Ozer HL, Banga SS, Dasgupta T, Houghton, J, Hubbard K, Jha KK, Kim SH, Lenahan M, Pang Z, Pardinas JR and Patsalis PC. . 1996 Exp. Gerontol. 31: 303–310.
Pai RB, Pai SB, Kukhanova M, Dutschman GE, Guo X and Cheng YC. . 1998 Cancer Res. 58: 1909–1913.
Pereira-Smith OM and Smith JR. . 1988 Proc. Natl. Acad. Sci. USA 85: 6042–6046.
Pitts AE and Corey DR. . 1998 Proc. Natl. Acad. Sci. USA 95: 11549–11554.
Pooga M, Soomets U, Hällbrink M, Valkna A, Saar K, Rezaei K, Kahl U, Hao J-X, Xu X-J, Wiesenfeld-Hallin Z, Hökfelt T, Bartfai T and Langel Ü. . 1998 Nature Biotechnol. 16: 857–861.
Reid LH, Shesely EG, Kim HS and Smithies O. . 1991 Mol. Cell. Biol. 11: 2769–2777.
Rubelj I, Venable SF, Lednicky J, Butel JS, Bilyeu T, Darlington G, Surmacz E, Campisi J and Pereira-Smith OM. . 1997 J. Gerontol. 52: B229–B234.
Sambrook J, Fritsch EF and Maniatis T. . (1989) Molecular Cloning; A Laboratory Manual Laboratory Press: Cold Spring Harbor, NY 9.47–9.58.
Sandhu AK, Hubbard K, Kaur GP, Jha KK, Ozer HL and Athwal RS. . 1994 Proc. Natl. Acad. Sci. USA 91: 5498–5502.
Shammas MA, Xia SJ, Shmookler Reis RJ. . 1997 Genetics 146: 1417–1428.
Sharma S, Raymond E, Soda H, Sun D, Hilsenbeck SG, Sharma A, Izbicka E, Windle B and Von Hoff DD. . 1997 Ann. Oncol. 8: 1063–1074.
Shaw JW and Bacchetti S. . 1997 Eur. J. Cancer 33: 787–791.
Shay JW and Wright WE. . 1989 Exp. Cell Res. 184: 109–118.
Shmookler Reis RJ and Goldstein S. . 1983 J. Biol. Chem. 258: 9078–9085.
Simmons CG, Pitts AE, Mayfield LD, Shay JW and Corey DR. . 1997 Bioorg. Med. Chem. Lett. 7: 3001–3006.
Smith JR, Pereira-Smith OM and Schneider EL. . 1978 Proc. Natl. Acad. Sci. USA 75: 1353–1356.
Wang S-S and Zakian VA. . 1990 Nature 345: 456–458.
Wheelhouse RT, Sun D, Han H, Han FX and Hurley LH. . 1998 J. Amer. Chem. Soc. 120: 3261–3262.
Wright WE, Pereira-Smith OM and Shay JW. . 1989 Mol. Cell. Biol. 9: 3088–3092.
Wright WE, Shay JW and Piatyszek MA. . 1995 Nucl. Acids Res. 23: 3794–3795.
Xia SJ, Shammas MA and Shmookler Reis RJ. . 1996 Mutat. Res. 364: 1–11.
Xia SJ, Shammas MA and Shmookler Reis RJ. . 1997 Mol. Cell. Biol. 17: 7151–7158.
Zakian VA. . 1997 Cell 91: 1–3.
Acknowledgements
This work was supported by grants from the U.S. Department of Veterans Affairs (to RJ Shmookler Reis), and the Robert R Welch Foundation (to DR Corey). DR Corey is an assistant investigator with the Howard Hughes Medical Institute.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shammas, M., Simmons, C., Corey, D. et al. Telomerase inhibition by peptide nucleic acids reverses `immortality' of transformed human cells. Oncogene 18, 6191–6200 (1999). https://doi.org/10.1038/sj.onc.1203069
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1203069
Keywords
This article is cited by
-
Artificial genetic polymers against human pathologies
Biology Direct (2022)
-
Targeting homologous recombination and telomerase in Barrett’s adenocarcinoma: impact on telomere maintenance, genomic instability and tumor growth
Oncogene (2014)
-
Genomic evolution in Barrett's adenocarcinoma cells: critical roles of elevated hsRAD51, homologous recombination and Alu sequences in the genome
Oncogene (2011)
-
Telomerase inhibitor GRN163L inhibits myeloma cell growth in vitro and in vivo
Leukemia (2008)
-
Telomerase inhibition by siRNA causes senescence and apoptosis in Barrett's adenocarcinoma cells: mechanism and therapeutic potential
Molecular Cancer (2005)