Abstract
A low proliferating fraction in solid tumors limits the effectiveness of cell cycle–dependent chemotherapeutic agents. To understand the molecular basis of such “kinetic” resistance we cultured tumor cells as multicellular spheroids and examined levels of p27kip1, a cyclin–dependent kinase inhibitor known to be upregulated by intercellular contact in normal cells. When transferred from monolayer to three–dimensional culture, a consistent upregulation (up to 15–fold) of p27 protein was observed in a panel of mouse and human carcinoma cell lines. Antisense–oligonucleotide–mediated downregulation of p27 in EMT–6 mammary tumor cell spheroids reduced intercellular adhesion, increased cell proliferation, sensitized tumor cells to 4–hydroperoxycyclophosphamide, and restored drug– or radiation–induced cell–cycle perturbations repressed in spheroid culture. Our results implicate p27 as a regulator of drug resistance in solid tumors and suggest that tumor–targeted p27 antagonists may be useful chemosensitizers in conjunction with conventional anticancer therapy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
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
Similar content being viewed by others
References
Tannock, I. Cell kinetics and chemotherapy: A critical review. Cancer Treat Rep. 62, 1117–1133 (1978).
Tannock, I.F. Principles of cell proliferation: Cell kinetics. in: Current Cancer Therapeutics. (eds. Kirkwood, J.M., Lotze, M.T. & Yasko, J.M.) 3–13 (Princeton Academic Press, Princeton, New Jersey, 1994).
Sutherland, R.M. Cell and environment interactions in tumor microregions: The multicell spheroid model. Science 240, 177–184 (1988).
Olive, P.L. & Durand, R.E. Drug and radiation resistance in spheroids Cell contact and kinetics. Cancer Metastasis Rev. 13, 121–138 (1994).
Teicher, B.A. et al. Tumor resistance to alkylating agents conferred by mechanisms operative only in vivo. Science 247, 1457–1461 (1990).
Kobayashi, H., Man, S., Kapitain, S.J., Teicher, B.A. & Kerbel, R.S. Acquired multicellular-mediated resistance to alkylating agents in cancer. Proc. Natl. Acad. Sci. USA 90, 3294–3298 (1993).
St Croix, B. et al. Reversal by hyaluronidase of adhesion-dependent multicellular drug resistance in mammary carcinoma cells. J. Natl. Cancer Inst. 88, 1285–1296 (1996).
Kerbel, R.S. et al. Multicellular resistance: A new paradigm to explain aspects of acquired drug resistance of solid tumors. Cold. Spring. Harbor. Symp. Quant. Biol. 59, 661–672 (1994).
Hoffman, R.M. The three-dimensional question: Can clinically relevant tumor drug resistance be measured in vitro? Cancer Metastasis Rev. 13, 169–173 (1994).
Morgan, D.O. Principles of CDK regulation. Nature 374, 131–134 (1995).
Sherr, C.J. & Roberts, J.M. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev. 9, 1149–1163
Hunter, T. & Pines, J. Cyclins and cancer. II. Cyclin D and CDK inhibitors come of age. Cell 79 573–582 (1994).
Ponce-Castaneda, M.V. et al p27kip1 : Chromosomal mapping to 12pl2-12pl3.1 and absence of mutations in human tumors. Cancer Res. 55, 1211–1214 (1995).
Kawamata, N. et al. Molecular analysis of the cyclin-dependent kinase inhibitor gene p27/Kip1 in human malignancies. Cancer Res. 55, 2266–2269 (1995).
Shiohara, M. et al. Absence of WAF1 mutations in a variety of human malignancies. Blood 84, 3781–3784 (1994).
Ferrando, A.A. et al. Mutational analysis of the human cyclin-dependent kinase inhibitor p27kip1in primary breast carcinomas. Hum Genet. 97, 91–94 (1996).
Spirin, K.S. et al. p27/Kip1 mutation found in breast cancer. Cancer Res. 56, 2400–2404 (1996).
Slingerland, J.M. et al. A novel inhibitor of cyclin-Cdk activity detected in transforming growth factor (β-arrested epithelial cells. Mol. Cell. Biol. 14, 3683–3694 (1994).
Hengst, L., Dulic, V., Slingerland, J.M., Lees, E. & Reed, S.I. A cell cycle-regulated inhibitor of cyclin-dependent kinases. Proc Natl. Acad. Sci. USA 91, 5291–5295 (1994).
Polyak, K. et al. p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-β and contact inhibition to cell cycle arrest. Genes Dev. 8, 9–22 (1994).
Wagner, R.W. et al. Antisense gene inhibition by oligonucleotides containing C-5 propyne pyrimidines. Science 260, 1510–1513 (1993).
Coats, S., Flanagan, W.M., Nourse, J. & Roberts, J.M. Requirement of p27Kip1 for restriction point control of the fibroblast cell cycle. Science 272 877–880 (1996).
Reyniskottir, I., Polyak, K. Iavarone, A. & Massague, J. Kip/Cip and Ink4 Cdk inhibitors cooperate to induce cell cycle arrest in response to TGF-β Genes Dev. 9 1831–1845 (1995).
Conte, P.F. et al. Conventional versus cytokinetic polychemotherapy with estrogenic recruitment in metastatic breast cancer: Results of a randomized cooperative trial. J. Clin. Oncol. 5, 339–347 (1987).
Gore, S.D. et al. Impact of in vivo administration of interleukin 3 on proliferation, differentiation, and chemosensitivity of acute myeloid leukemia. Clin. Cancer Res. 1, 295–303 (1995).
Karp, J.E. & Broder, S. Molecular foundations of cancer: New targets for intervention. Nature Med. 1, 309–320 (1995).
Fang, F., Orend, G., Watanabe, N., Hunter, T. & Ruoslahti, E. Dependence of cyclin E-Cdk2 kinase activity on cell anchorage. Science 271, 499–502 (1996).
Sano, Y., Hoshino, T., Bjerkvig, R. & Deen, D.F. The relative resistance of non-cycling cells in 9L multicellular spheroids to spirohydantoin mustard. Eur. J. Cancer Clin. Oncol. 19, 1451–1456 (1983).
Hinz, G. & Dertinger, H. Increased radioresistance of cells in cultured multicell spheroids. Radiat Environ Biophys. 21, 255–264 (1983).
Remvikos, Y. et al. Cell cycle modifications of breast cancers during neoadjuvant chemotherapy: A flow cytometry study on fine needle aspirates. Eur. J. Cancer 29A 1843–1848 (1993).
Briffod, M. et al. Evaluation of breast carcinoma chemosensitivity by flow cyto-metric DNA analysis and computer assisted image analysis. Cytometry 13, 250–258 (1992).
Spyratos, F. et al. Sequential cytopunctures during preoperative chemotherapy for primary breast carcinoma. Cancer 69, 470–475 (1992).
Meredith, J.E. Jr., Fazeli, B. & Schwartz, M.A. The extra cellular matrix as a cell survival factor. Mol. Biol. Cell 4, 953–961 (1993).
Frisch, S.M. & Francis, H. Disruption of epithelial cell-matrix interactions induces apoptosis. J. Cell. Rial. 124, 619–626 (1994).
Bates, R.C., Buret, A. van Helden, D.F., Horton, M.A. & Burns, G.F. Apoptosis induced by inhibition of inter cellular contact. J. Cell Biol. 125, 403–415 (1994).
Mendonca, M.S., Rodriguez, A. & Alpen, E.L. Differential repair of potentially lethal damage in exponentially growing and quiescent 9L Cells. Rad. Res. 122, 38–43 (1990).
Little, J.B., Hahn, G.M., Frindel, E. & Tubiana, M. Repair of potentially lethal radiation damage in vitro and in vivo . Radiology 106, 689–694 (1973).
Dimanche-Boitrel, M.-T., Garrido, C. & Chauffert, B. Kinetic resistance to anticancer agents. Cytotechnology 12, 347–356 (1993).
Dimanche-Boitrel, M. et al. Confluence-dependent resistance in human colon cancer cells: Role of reduced drug accumulation and low intrinsic chemosensitivity of resting cells. Int. J. Cancer 50, 677–682 (1992).
Zhang, W. et al. High levels of constitutive WAFl/Cipl protein are associated with chemoresistance in acute myelogenous leukemia. Clin. Cancer Res. 1, 1051–1057 (1995).
Poluha, W. et al. The cyclin-dependent kinase inhibitor p21WAF1 is required for survival of differentiating neuroblastoma cells. Mol. Cell. Biol. 16, 1335–1341 (1996).
Waldman, T., Lengauer, C., Kinzler, K.W. & Vogelstein, B. Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21. Nature 381, 713–716 (1996).
Wang, J. & Walsh, K. Resistance to apoptosis conferred by Cdk inhibitors during myocyte differentiation. Science 273, 359–361 (1996).
Deng, C., Zhang, P., Harper, J.W., Elledge, S.J. & Leder, P. Mice lacking p21CIP1/WAF1undergo normal development, but are defective in Gl checkpoint control. Cell 82, 675–684 (1995).
Nakayama, K. et al. Mice lacking p27kip1display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 85, 707–720 (1996).
Kiyokawa, H. et al. Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27kip1 . Cell 85, 721–732 (1996).
Fero, M.L. et al. A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27kip1-deficient mice. Cell 85, 733–744 (1996).
Tonkinson, J.L. & Stein, C.A. Antisense oligodeoxynucleotides as clinical therapeutic agents. Cancer Invest. 14 (1), 54–65 (1996).
Hietanen, P. et al. Do DNA ploidy and S-phase fraction in primary tumour predict the response to chemotherapy in metastatic breast cancer. Br. J. Cancer 71, 1029–1032 (1995).
O'Reilly, S.M., Camplejohn, R.S., Rubens, R.D. & Richards, M.A. DNA flow cytometry and response to preoperative chemotherapy for primary breast cancer. Eur. J. Cancer 213, 681–683 (1992).
Remvikos, Y. et al. Correlation of pretreatment proliferative activity of breast cancer with the response to cytotoxic chemotherapy. J. Natl. Cancer Inst. 81, 1383–1387 (1989).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Croix, B., Flørenes, V., Rak, J. et al. Impact of the cyclin–dependent kinase inhibitor p27Kip1 on resistance of tumor cells to anticancer agents. Nat Med 2, 1204–1210 (1996). https://doi.org/10.1038/nm1196-1204
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nm1196-1204
This article is cited by
-
A phase I study of the safety and tolerability of VLX600, an Iron Chelator, in patients with refractory advanced solid tumors
Investigational New Drugs (2019)
-
PI3K/AKT pathway-mediated regulation of p27Kip1 is associated with cell cycle arrest and apoptosis in cervical cancer
Cellular Oncology (2015)
-
Contribution of very late antigen-4 (VLA-4) integrin to cancer progression and metastasis
Cancer and Metastasis Reviews (2015)
-
Induction of mitochondrial dysfunction as a strategy for targeting tumour cells in metabolically compromised microenvironments
Nature Communications (2014)
-
Hepatocyte induced re-expression of E-cadherin in breast and prostate cancer cells increases chemoresistance
Clinical & Experimental Metastasis (2012)