Abstract
Pancreatic cancers frequently carry mutations in the Kras, p53, and p16 genes, which regulate cell proliferation. Transition from G1 to S phase of the cell cycle requires activation of cyclin-dependent kinase 2 (Cdk2), which is inhibited by olomoucine and roscovitine. The purpose of this study was to determine whether olomoucine and roscovitine can block Cdk2 kinase activity and inhibit proliferation of four human pancreatic cancer cell lines with various genetic alterations. Human pancreatic carcinoma cell lines BxPC-3, PANC-1, Capan-2, and CAV were treated with olomoucine or roscovitine. Cdk2 kinase activity was determined using histone H1 as the substrate. Cell cycle distribution was analyzed by DNA flow cytometry. Cell numbers were quantitated by Coulter counter. Olomoucine and roscovitine blocked Cdk2 activity in all four pancreatic cancer cell lines. Both compounds also inhibited cell proliferation in a dose-dependent fashion. Roscovitine was at least threefold more potent than olomoucine for both Cdk2 activity and cell proliferation. We have shown that Cdk inhibitors, olomoucine and roscovitine, block proliferation of human pancreatic cancer cells regardless of their mutations in Kras, p53, or pl 6 genes. These compounds represent a novel therapeutic strategy with potential therapeutic benefits for pancreatic cancers.
Similar content being viewed by others
References
Parker SL, Tong T, Bolden S, Wingo PA. Cancer Statistics, 1997. CA CancerJ Clin 1997;47:5–27.
Reber HA, Ashley SW, McFadden D. Curative treatment for pancreatic neoplasms. Radical resection. Surg Clin North Am 1995;75:905–912.
Abrams RA, Grochow LB. Adjuvant therapy with chemotherapy and radiation therapy in the management of carcinoma of the pancreatic head. Surg Clin North Am 1995;75:925–938.
Martin J, Arnheim N, Perucho M. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell 1988;53:549–554.
Kalthoff H, Schmiegel W, Roeder C, Kasche D, Schmidt A, Lauer G, Thiele HG, Honold G, Pantel K, Riethmuller G, Scherer E, Maurer J, Maacke H, Deppert W. p53 and K-ras alterations in pancreatic epithelial cell lesions. Oncogene 1993;8:289–298.
Pellegata NS, Sessa E Renault B, Bonato M, Leone BE, Solcia E, Ranzani GN. K-ras and p53 gene mutations in pancreatic cancer: Ductal and nonductal tumors progress through different genetic lesions. Cancer Res 1994;54:1556–1560.
Redson MS, Caldas C, Seymour AB, Hruban RH, da Costa L, Yeo CJ, Kern SE. p53 mutations in pancreatic carcinoma and evidence of common involvement of homocopolymer tracts in DNA microdeletions. Cancer Res 1994;54:3025–3033.
Caldas C, Hahn SA, da Costa LT, Redston MS, Schutte M, Seymour AB, Weinstein CL, Hruban RH, Yeo CJ, Kern SE. Frequent somatic mutations and homozygous deletions of the pl6 (MTS1) gene in pancreatic adenocarcinoma. Nature Genet 1994;8:27–32.
van den Heuvel S, Harlow E. Distinct roles for cyclin-dependent kinases in cell cycle control. Science 1993;262:2050–2054.
Nigg EA. Cyclin-dependent protein kinases: Key regulators of the eukaryotic cell cycle. Bioessays 1995;17:471–480.
Hunter T, Pines J. Cyclins and cancer. Cell 1991;66:1071–1074.
Sherr CJ, Roberts JM. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev 1995;9:1149–1163.
Elledge SJ, Harper JW. Cdk inhibitors: On the threshold of checkpoints and development. Curr Opin Cell Biol 1994;6:847–852.
Nurse P. Ordering S phase and M phase in the cell cycle. Cell 1994;79:547–550.
Dulic V, Lees E, Reed SI. Association of human cyclin E with a periodic G1-S phase protein kinase. Science 1992;257:1958–1961.
Ohtsubo M, Theodoras AM, Schumacher J, Roberts JM, Pagano M. Human cyclin E, a nuclear protein essential for the Gl-to-S phase transition. Mol Cell Biol 1995;15:2612–2624.
Serrano M, Hannon G, Beach D. A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature 1993;366:704–707.
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. The p21 Cdk-interacting protein Cipl is a potent inhibitor of G1 cyclin-dependent kinases. Cell 1993;75:805–816.
E1-Deiry W, Tokino T, Velculescu VE, Levy DB, Parson VE, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B. WAF1, a potential mediator of p53 tumour suppression. Cell 1993;75:817–825.
E1-Deiry WS, Harper JW, O’Connor PM, Velculescu V, Canman CE, Jackman J, Pietenpol J, Burrell M, Hill DE, Wiman KG, Mercer WE, Kastan MB, Kohn KW, Elledge SJ, Kinzler KW, Vogelstein B. WAF1/CIP1 is induced in p53 mediated G1 arrest and apoptosis. Cancer Res 1994;54:1169–1174.
Vesely J, Havlicek L, Strnad M, Blow JJ, Donella-Deana A, Pinna L, Letham DS, Kato JY, Detivaud L, LeClerc S, Meijer L. Inhibition of cyclin-dependent kinases by purine analogues. EurJ Biochem 1994;224:771–786.
Meijer L. Chemical inhibitors of cyclin-dependent kinases. Trends Cell Biol 1996;6:393–397.
Iseki H, Ko TC, Xue XY, Seapan A, Hellmich MR, Townsend CM Jr. Cyctin-dependent kinase inhibitors block proliferation of human gastric cancer cells. Surgery 1997; 122:187 -195.
Abraham RT, Acquarone M, Andersen A, Asensi A, Belle R, Berger F, Bergounioux C, Brunn G, Buquet-Fagot C, Fagot D, Glab N, Goudeau H, Goudeau M, Guerrier P, Houghton P, Hendriks H, Kloareg B, Lippai M, Marie D, Maro B, Mei-jer L, Mester J, Mulner-Lorillon O, Poulet SA, Schierenberg E, Schutte B, Vaulot D, Verlhac MH. Cellular effects of olomoucine, an inhibitor of cyclin-dependent kinases. Biol Cell 1995;83:105–120.
Meijer L, Borgne A, Mulner O, Chong JP, Blow JJ, Inagaki N, Inagaki M, Delcros JG, Moulinoux JP. Biochemical and cellular effects of roscovitine, a potent and selective inhibitor of the cyclin-dependent ldnases cdc2, cdk2 and cdkS. Eur J Biochem 1997;243:527–536.
Upp JR Jr, Olson D, Poston GJ, Alexander RW, Townsend CM Jr, Thompson JC. Inhibition of growth of two human pancreatic adenocarcinomas in vivo by somatostatin analog SMS 201-995. AmJ Surg 1988;155:29–35.
Saydgari R, Alexander RW, Upp JR Jr, Barranco SC, Townsend CM Jr, Thompson JC. Differential sensitivity of various human tumors to inhibition of polyamine biosynthesis in vivo. IntJ Cancer 1991;47:44–48.
Evers BM, Ko TC, Li J, Thompson EA. Cell cycle protein suppression and p21 induction in differentiating Caco-2 cells. Am J Physiol 1996;271:G722-G727.
Aold K, Yoshida T, Sugimua T, Terada M. Liposome-mediated in vivo gene transfer of antisense K-ras construct inhibits pancreatic tumor dissemination in the murine peritoneal cavity. Cancer Res 1995;55:3810–3816.
Barton CM, Lemoine NR. Antisense oligonucleotides directed against p53 have antiproliferative effects unrelated to effects on p53 expression. BrJ Cancer 1995;71:429–437.
Berrozpe G, Schaeffer J, Peinado MA, Real FX, Perucho M. Comparative analysis of mutations in the p53 and K-ras genes in pancreatic cancer. IntJ Cancer 1994;58:185–191.
Barton CM, Staddon SL, Hughes CM, Hall PA, O’Sullivan C, Kloppel G, Theis B, Russell RCG, Neoptolemos J, Williamson RCN, Lane DP, Lemoine NR. Abnormalities of the p53 tumour suppressor gene in human pancreatic cancer. Br J Cancer 1991;64:1076–1082.
Naumann M, Savitskaia N, Eilert C, Schramm A, Kalthoff H, Schmiegel W. Frequent codeletion of pl6/MTS1 and pl5/MTS2 and genetic alterations in pI6/MTS1 in pancreatic tumors. Gastroenterology 1996; 110:1215 -1224.
Morgan DO. Principles of CDK regulation. Nature 1995; 374:131–134.
Lew DJ, Kornbluth S. Regulatory roles of cyclin-dependent kinase phosphorylation in cell cycle control. Curr Opin Cell Biol 1996;8:795–804.
Parker CW, Entsch B, Letham DS. Inhibitors of two enzymes which metabolize cytokinins. Phytochemistry 1986;25:303–310.
Author information
Authors and Affiliations
Additional information
Supported by grants from the National Institutes of Health (K08 CA64191, R01 DK48345), the John S ealy Memorial Endowment Fund, and the Walls Medical Research Foundation.
Rights and permissions
About this article
Cite this article
Iseki, H., Ko, T.C., Xue, X.Y. et al. A novel strategy for inhibiting growth of human pancreatic cancer cells by blocking cyclin-dependent kinase activity. J Gastrointest Surg 2, 36–43 (1998). https://doi.org/10.1016/S1091-255X(98)80101-7
Issue Date:
DOI: https://doi.org/10.1016/S1091-255X(98)80101-7