Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Comparison of the effectiveness of adenovirus vectors expressing cyclin kinase inhibitors p16INK4A, p18INK4C, p19INK4D, p21WAF1/CIP1 and p27KIP1 in inducing cell cycle arrest, apoptosis and inhibition of tumorigenicity

Abstract

Cell cycle regulatory proteins are important candidates for therapeutic tumour suppressors. Adenovirus vectors were constructed to overexpress cyclin kinase inhibitors p16INK4A, p18INK4C, p19INK4D, p21WAF1/CIP1 and p27KIP1 under the control of the murine cytomegalovirus immediate early gene promoter. These vectors directed the efficient expression of each of the cyclin kinase inhibitors and induced growth arrest, inhibited DNA synthesis, and prevented phosphorylation of the retinoblastoma protein (pRb) in cell lines expressing functional pRb. In pRb-deficient cells, expression of the cyclin kinase inhibitors was not effective in inhibiting DNA replication or growth arrest. Interestingly, three of the cyclin kinase inhibitors, p16, p18 and p27 were found to induce apoptotic death in transduced HeLa and A549 cells. When the vectors were tested for their ability to inhibit tumorigenicity in a polyomavirus middle T antigen model of murine breast carcinoma, expression of the cyclin kinase inhibitors resulted in a delay in tumour formation that varied from several weeks for the p19 expressing vector to greater than 25 weeks for the p27 expressing vector. When tumours were injected directly with the adenovirus vectors expressing the cyclin kinase inhibitors, only treatment with the vector expressing p16 resulted in a delay in tumour growth.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

Abbreviations

Ad:

adenovirus

CDK:

cyclin dependent kinase

CKI:

cyclin dependent kinase inhibitor

h.r.p.:

horseradish peroxidase

m.o.i.:

multiplicity of infection

PBS:

phosphate buffered saline

p.f.u.:

plaque forming unit

pRb:

retinoblastoma protein

PyMidT:

polyoma virus middle T antigen

References

  • Adachi M, Roussel MF, Havenith K and Sherr CJ. . 1997 Blood 90: 126–137.

  • Addison CL, Braciak T, Ralston R, Muller W, Gauldie J and Graham FL. . 1995 Proc. Natl. Acad. Sci. USA 92: 8522–8526.

  • Addison CL, Hitt M, Kunsken D and Graham F. . 1997 J. Gen. Virol. 78: 1653–1661.

  • Albert ML, Sauter B and Bhardwaj N. . 1998 Nature 392: 86–89.

  • Bacchetti S and Graham F. . 1993 Int. J. Oncol. 3: 781–788.

  • Bett AJ, Haddara W, Prevec L and Graham FL. . 1994 Proc. Natl. Acad. Sci. USA 91: 8802–8806.

  • Cayrol C, Knibiehler M and Ducommum B. . 1998 Oncogene 16: 311–320.

  • Chellappan S, Kraus VB, Kroger B, Munger K, Howley PM, Phelps WC and Nevins JR. . 1992 Proc. Natl. Acad. Sci. USA 89: 4549–4553.

  • Chow VT, Quek HH and Tock EP. . 1993 Cancer Lett. 73: 141–148.

  • Cox LS. . 1997 J. Pathol. 183: 134–140.

  • Craig C, Kim M, Ohri E, Wersto R, Katayose D, Li A, Choi YH, Madahar B, Srivastava S, Seth P and Cowan K. . 1998 Oncogene 16: 265–272.

  • Craig C, Wersto R, Kim M, Ohri E, Li Z, Katayose D, Lee SJ, Trepel J, Cowan K and Seth P. . 1997 Oncogene 14: 2283–2289.

  • Deng C, Zhang P, Harper W, Elledge SJ and Leder P. . 1995 Cell 82: 675–684.

  • Diller L, Kassel J, Nelson C, Gryka M, Litwak G, Gebhardt M, Bressac B, Ozturk M, Baker S, Vogelstein B and Friend S. . 1990 Mol. Cell. Biol. 10: 5772–5781.

  • Eastham JA, Hall SJ, Sehgal I, Wang J, Timme T, Yang G, Connell-Crowley L, Elledge SJ, Zhang W-W, Harper JW and Thompson TC. . 1995 Cancer Res. 55: 5151–5155.

  • Elledge SJ, Winston J and Harper JW. . 1996 Trends Cell Biol. 6: 388–392.

  • Fero ML, Rivkin M, Tasch M, Porter P, Carow C, Firpo E, Polyak K, Tsai L-H, Broudy V, Perimutter RM, Kaushansky K and Roberts JM. . 1996 Cell 85: 733–744.

  • Gartel A, Serfas M and Tyner AL. . 1996 Proc. Soc. Exp. Biol. Med. 213: 138–149.

  • Gorospe M, Cirielli C, Wang X, Seth P, Capogrossi MC and Holbrook NJ. . 1997 Oncogene 14: 929–935.

  • Gotoh A, Kao C, Ko S-C, Hamada K, Liu T-J and Chung L. . 1997 J. Urol. 158: 636–641.

  • Grana X and Reddy EP. . 1995 Oncogene 11: 211–219.

  • Hamada K, Alemany R, Zhang W-W, Hittelman W, Lotan R, Roth JA and Mitchell MF. . 1996 Cancer Res. 56: 3047–3054.

  • Harper JW and Elledge SJ. . 1996 Curr. Opin. Genet. Dev. 6: 56–64.

  • Herwig S and Strauss M. . 1997 Eur. J. Biochem. 246: 581–601.

  • Hirai H, Roussel MF, Kato J-Y, Ashmun RA and Sherr CJ. . 1995 Mol. Cell Biol. 15: 2672–2681.

  • Hitt M, Bett AJ, Addison CL, Prevec L and Graham FL. . 1995 Meth. Mol. Genet. 7: 13–30.

  • Hitt M, Bett AJ, Prevec L and Graham FL. . 1994 Cell Biology: A Laboratory Handbook. Celis, J.E. (ed.). Academic Press: San Diego pp. 479–90.

    Google Scholar 

  • Hoffman B and Lieberman DA. . 1994 Oncogene 9: 1807–1812.

  • Hunter T and Pines J. . 1994 Cell 79: 573–582.

  • Jiang H, Lin J, Su Z-s, Collart FR, Huberman E and Fisher PB. . 1994 Oncogene 9: 3397–3406.

  • Jin X, Nguyen D, Zhang W-W, Kyritsis AP and Roth JA. . 1995 Cancer Res. 55: 3250–3253.

  • Johnson P, Gray D, Mowat M and Benchimol S. . 1991 Mol. Cell Biol. 11: 1–11.

  • Kamb A. . 1995 Trends Genet. 11: 136–140.

  • Katayose D, Wersto R, Cowan K and Seth P. . 1998 Cell Growth Differ. 6: 1207–1212.

  • Katayose Y, Kim M, Rakkar ANS, Li Z, Cowan KH and Seth P. . 1997 Cancer Res. 57: 5441–5445.

  • Kaufmann S, Desnoyers S, Ottaviano Y, Davidson NE and Poirier G. . 1993 Cancer Res. 53: 3976–3985.

  • Kawakami K, Futami H, Takahara J and Yamaguchi K. . 1996 Biochem. Biophys. Res. Commun. 219: 778–783.

  • Kiyokawa H, Kineman RD, Manova-Todorova K, Soares VC, Hoffman ES, Ono M, Khanam D, Hayday AC, Frohman LA and Koff A. . 1996 Cell 85: 721–732.

  • Liebermann DA, Hoffman B and Steinman RA. . 1995 Oncogene 11: 199–210.

  • Lu Y, Yamagishi N, Yagi I and Takebe H. . 1998 Oncogene 16: 705–712.

  • Lukas J, Muller H, Bartkova J, Spitkovsky D, Kjerulff A, Jansen-Durr P, Strauss M and Bartek J. . 1994 J. Cell Biol. 125: 625–638.

  • Mayol X and Grana X. . 1997 Prog. Cell Cycle Res. 3: 157–169.

  • Morgan DO. . 1995 Nature 374: 131–134.

  • Morse L, Chen D, Franklin D, Xiong Y and Chen-Kiang S. . 1997 Immunity 6: 47–56.

  • Mulligan G and Jacks T. . 1998 Trends Genet. 14: 223–229.

  • Nakayama K, Ishida N, Shirane M, Iomata A, Inoue T, Horii SI, Loh DY and Nakayama K-i. . 1996 Cell 85: 707–720.

  • Niculescu ABI, Chen X, Smeets M, Hengst L, Prives C and Reed S. . 1998 Mol. Cell. Biol. 18: 629–643.

  • Ohno T, Yang Z, Ling X, Jaffe M, Nabel EG, Normolle D and Nabel GJ. . 1997 Gene Ther. 4: 361–366.

  • O'Malley BW, Cope KA, Chen S-H, Li D, Schwartz MR and Woo SLC. . 1996 Cancer Res. 56: 1737–1741.

  • Putzer B, Bramson JL, Addison CL, Hitt M, Siegel PM, Muller WJ and Graham F. . 1998 Hum. Gene Ther. 9: 707–718.

  • Sambrook J, Fritsch EF and Maniatis T. (1989). . Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

  • Sandig V, Brand K, Herwig S, Lukas J, Bartek J and Strauss M. . 1997 Nature Med. 3: 313–319.

  • Schwaller J, Pabst T, Koeffler HP, Niklaus G, Loestcher P, Fey MF and Tobler A. . 1997 Leukemia 11: 54–63.

  • Sellers WR and Kaelin WGJ. . 1997 J. Clin. Oncol. 15: 3301–3312.

  • Serrano M, Lee H-W, Chin L, Cordon-Cardo C, Beach D and DePinho RA. . 1996 Cell 85: 27–37.

  • Sgonc R, Boeck G, Dietrich H, Gruber J, Recheis H and Wick G. . 1994 Trends Genet. 10: 41–42.

  • Sherr C. . 1996 Science 274: 1672–1677.

  • Sherr CJ. . 1994 Cell 79: 551–555.

  • Sherr CJ and Roberts JM. . 1995 Genes Dev. 9: 1149–1163.

  • Shew JY, Lin BT, Chen PL, Tseng BY, Yang-Feng TL and Lee WH. . 1990 Proc. Natl. Acad. Sci. USA. 87: 6–10.

  • Steinman RA, Hoffman B, Iro A, Guillouf C, Liebermann DA and El-Houseini ME. . 1994 Oncogene 9: 3389–3396.

  • Terada Y, Yamada T, Nakashima O, Tamamori M, Ito H, Sasaki S and Marumo F. . 1997 J. Am. Soc. Nephrol. 8: 51–60.

  • Tewari M, Quan LT, O'Rourke K, Desnoyers S, Zeng Z, Beider DR, Poirier G, Salvesen GS and Dixit V. . 1995 Cell 81: 801–809.

  • Urashima M, DeCaprio JA, Chauhan D, Teoh G, Ogata A, Treon SP, Hoshi Y and Anderson KC. . 1997 Blood 90: 4106–4115.

  • Wang Z, Gorospe M, Huang Y and Holbrook NJ. . 1997 Oncogene 15: 2991–2997.

  • Warneford S, Townsend M, Rowe PB, Dalla-Pozza L and Symonds G. . 1991 Cell Growth Differ. 2: 439–445.

  • Weinbert RA. . 1995 Cell 81: 323–330.

  • Xiong Y. . 1996 Biochim. Biophys. Acta 1288: O1–O5.

  • Yang Z-Y, Perkins N, Ohno T, Nabel EG and Nabel GJ. . 1995 Nature Med. 1: 1052–1056.

  • Zuo Z, Dean NM and Honkanen RE. . 1998 J. Biol. Chem. 273: 12250–12258.

Download references

Acknowledgements

We thank J Rudy and U Sankar for excellent technical assistance, R Marr for preparing PyMidT primary cells and mouse injections, Inger Bengtsson for assistance with cell cycle analysis and Dr M Hitt for helpful discussions and critically reading the manuscript. This work was supported by grants from the National Cancer Institute of Canada (NCIC) and the Medical Research Council of Canada (MRC). FLG is a Terry Fox scientist of the NCIC, WJM is a MRC scholar and MS is supported by a MRC postdoctoral fellowship. All animal work has been approved and carried out according to guidelines set by the Animal Research Ethics Board of McMaster University.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schreiber, M., Muller, W., Singh, G. et al. Comparison of the effectiveness of adenovirus vectors expressing cyclin kinase inhibitors p16INK4A, p18INK4C, p19INK4D, p21WAF1/CIP1 and p27KIP1 in inducing cell cycle arrest, apoptosis and inhibition of tumorigenicity. Oncogene 18, 1663–1676 (1999). https://doi.org/10.1038/sj.onc.1202466

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1202466

Keywords

This article is cited by

Search

Quick links