Cell Growth & Differentiation, Vol 6, Issue 8 927-936, Copyright © 1995 by American Association of Cancer Research

ARTICLES

Apoptosis in 7-hydroxystaurosporine-treated T lymphoblasts correlates with activation of cyclin-dependent kinases 1 and 2 [published erratum appears in Cell Growth Differ 1995 Oct;6(10):1339]

Q Wang, PJ Worland, JL Clark, BA Carlson and EA Sausville
Laboratory of Biological Chemistry, Division of Cancer Treatment, National Cancer Institute, Bethesda, Maryland 20892, USA.

7-Hydroxystaurosporine (UCN-01) is a potent inhibitor of protein kinase C (PKC) isozymes alpha, beta, and gamma [Seynaeve et al., Mol. Pharmacol, 45: 1207-1214, 1994] that also has antitumor effects in vivo. To determine whether inhibition of PKC can be related to inhibition of cell growth with induction of apoptosis, we compared the effects of UCN-01 to those of the highly selective bisindolylmaleimide PKC antagonist GF 109203X in leukemic T-cell lines. Both compounds potently inhibited PKC activity when added to T-cell membrane preparations and reversed phorbol ester-induced c-fos gene expression in intact cells. However, whereas UCN-01 potently inhibited growth of Jurkat, Molt-3, Molt-4, and Hut-78 cells (IC50 = 20-65 nM, irreversible after 24 h of exposure), GF 109203X had IC50s for cell growth of 3.6-5.0 muM. Less than 3 h after addition, UCN-01 but not GF 109203X-treated cells displayed loss of cells with G2-M DNA content, appearance of a hypodiploid DNA fraction, and evidence of internucleosomal DNA fragmentation. Six h after treatment, cells appeared to accumulate with S-phase DNA content. These effects correlated with selective UCN-01 but not GF 109203X-induced decrease in total and tyrosine phosphorylation of cyclin-dependent kinases (cdks) 1 and 2, and with increases in the histone H1 kinase activities of cdk1 and cdk2. UCN-01 was relatively less potent in inhibition of properly activated cdk1 and cdk2 when added in vitro to H1 kinase assays (IC50 = 1000 and 600 nM, respectively). We conclude that inhibition of PKC alone is not sufficient to account for the actions of UCN-01 and are led to the hypothesis that inappropriate cdk activation either correlates with or actually mediates cell growth inhibition with apoptosis in T lymphoblasts exposed to UCN-01.

This article has been cited by other articles:

				H. Hamed, W. Hawkins, C. Mitchell, D. Gilfor, G. Zhang, X.-Y. Pei, Y. Dai, M. P. Hagan, J. D. Roberts, A. Yacoub, et al. Transient exposure of carcinoma cells to RAS/MEK inhibitors and UCN-01 causes cell death in vitro and in vivo Mol. Cancer Ther., March 1, 2008; 7(3): 616 - 629. [Abstract] [Full Text] [PDF]

				T. Hattori, T. Isobe, K. Abe, H. Kikuchi, K. Kitagawa, T. Oda, C. Uchida, and M. Kitagawa Pirh2 Promotes Ubiquitin-Dependent Degradation of the Cyclin-Dependent Kinase Inhibitor p27Kip1 Cancer Res., November 15, 2007; 67(22): 10789 - 10795. [Abstract] [Full Text] [PDF]

				B. Ewald, D. Sampath, and W. Plunkett H2AX phosphorylation marks gemcitabine-induced stalled replication forks and their collapse upon S-phase checkpoint abrogation Mol. Cancer Ther., April 1, 2007; 6(4): 1239 - 1248. [Abstract] [Full Text] [PDF]

				X.-Y. Pei, W. Li, Y. Dai, P. Dent, and S. Grant Dissecting the Roles of Checkpoint Kinase 1/CDC2 and Mitogen-Activated Protein Kinase Kinase 1/2/Extracellular Signal-Regulated Kinase 1/2 in Relation to 7-Hydroxystaurosporine-Induced Apoptosis in Human Multiple Myeloma Cells Mol. Pharmacol., December 1, 2006; 70(6): 1965 - 1973. [Abstract] [Full Text] [PDF]

				T. Takahashi, F. Yamasaki, T. Sudo, H. Itamochi, S. Adachi, M. Tamamori-Adachi, and N. T. Ueno Cyclin A-associated kinase activity is needed for paclitaxel sensitivity Mol. Cancer Ther., July 1, 2005; 4(7): 1039 - 1046. [Abstract] [Full Text] [PDF]

				X.-Y. Pei, Y. Dai, M. Rahmani, W. Li, P. Dent, and S. Grant The Farnesyltransferase Inhibitor L744832 Potentiates UCN-01-Induced Apoptosis in Human Multiple Myeloma Cells Clin. Cancer Res., June 15, 2005; 11(12): 4589 - 4600. [Abstract] [Full Text] [PDF]

				E. A. Sausville Indifferently Pursued or Unowned Drugs: Who Should Lead Where Companies Do Not Tread? J. Clin. Oncol., March 20, 2005; 23(9): 1796 - 1798. [Full Text] [PDF]

				J. Kortmansky, M. A. Shah, A. Kaubisch, A. Weyerbacher, S. Yi, W. Tong, R. Sowers, M. Gonen, E. O'Reilly, N. Kemeny, et al. Phase I Trial of the Cyclin-Dependent Kinase Inhibitor and Protein Kinase C Inhibitor 7-Hydroxystaurosporine in Combination With Fluorouracil in Patients With Advanced Solid Tumors J. Clin. Oncol., March 20, 2005; 23(9): 1875 - 1884. [Abstract] [Full Text] [PDF]

				M. Hahn, W. Li, C. Yu, M. Rahmani, P. Dent, and S. Grant Rapamycin and UCN-01 synergistically induce apoptosis in human leukemia cells through a process that is regulated by the Raf-1/MEK/ERK, Akt, and JNK signal transduction pathways Mol. Cancer Ther., March 1, 2005; 4(3): 457 - 470. [Abstract] [Full Text] [PDF]

				E. C. Dees, S. D. Baker, S. O'Reilly, M. A. Rudek, S. B. Davidson, C. Aylesworth, K. Elza-Brown, M. A. Carducci, and R. C. Donehower A Phase I and Pharmacokinetic Study of Short Infusions of UCN-01 in Patients with Refractory Solid Tumors Clin. Cancer Res., January 15, 2005; 11(2): 664 - 671. [Abstract] [Full Text] [PDF]

				G. P. Dasmahapatra, P. Didolkar, M. C. Alley, S. Ghosh, E. A. Sausville, and K. K. Roy In vitro Combination Treatment with Perifosine and UCN-01 Demonstrates Synergism against Prostate (PC-3) and Lung (A549) Epithelial Adenocarcinoma Cell Lines Clin. Cancer Res., August 1, 2004; 10(15): 5242 - 5252. [Abstract] [Full Text] [PDF]

				M. M. Facchinetti, A. De Siervi, D. Toskos, and A. M. Senderowicz UCN-01-Induced Cell Cycle Arrest Requires the Transcriptional Induction of p21waf1/cip1 by Activation of Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Kinase/Extracellular Signal-Regulated Kinase Pathway Cancer Res., May 15, 2004; 64(10): 3629 - 3637. [Abstract] [Full Text] [PDF]

				M. Huigsloot, R. B. Tijdens, and B. van de Water Inhibition of Protein Kinase C{alpha} Enhances Anticancer Agent-Induced Loss of Anchorage-Independent Growth Regardless of Protection against Apoptosis by Bcl-2 Mol. Pharmacol., October 1, 2003; 64(4): 965 - 973. [Abstract] [Full Text] [PDF]

				W. Jia, C. Yu, M. Rahmani, G. Krystal, E. A. Sausville, P. Dent, and S. Grant Synergistic antileukemic interactions between 17-AAG and UCN-01 involve interruption of RAF/MEK- and AKT-related pathways Blood, September 1, 2003; 102(5): 1824 - 1832. [Abstract] [Full Text] [PDF]

				E. A. Kohn, C. J. Yoo, and A. Eastman The Protein Kinase C Inhibitor Go6976 Is a Potent Inhibitor of DNA Damage-induced S and G2 Cell Cycle Checkpoints Cancer Res., January 1, 2003; 63(1): 31 - 35. [Abstract] [Full Text] [PDF]

				V. Patel, T. Lahusen, C. Leethanakul, T. Igishi, M. Kremer, L. Quintanilla-Martinez, J. F. Ensley, E. A. Sausville, J. S. Gutkind, and A. M. Senderowicz Antitumor Activity of UCN-01 in Carcinomas of the Head and Neck Is Associated with Altered Expression of Cyclin D3 and p27KIP1 Clin. Cancer Res., November 1, 2002; 8(11): 3549 - 3560. [Abstract] [Full Text] [PDF]

				Y. Dai, T. H. Landowski, S. T. Rosen, P. Dent, and S. Grant Combined treatment with the checkpoint abrogator UCN-01 and MEK1/2 inhibitors potently induces apoptosis in drug-sensitive and -resistant myeloma cells through an IL-6-independent mechanism Blood, October 16, 2002; 100(9): 3333 - 3343. [Abstract] [Full Text] [PDF]

				A. M. Senderowicz The Cell Cycle as a Target for Cancer Therapy: Basic and Clinical Findings with the Small Molecule Inhibitors Flavopiridol and UCN-01 Oncologist, August 1, 2002; 7(90003): 12 - 19. [Abstract] [Full Text] [PDF]

				T. Yamauchi, M. J. Keating, and W. Plunkett UCN-01 (7-Hydroxystaurosporine) Inhibits DNA Repair and Increases Cytotoxicity in Normal Lymphocytes and Chronic Lymphocytic Leukemia Lymphocytes Mol. Cancer Ther., February 1, 2002; 1(4): 287 - 294. [Abstract] [Full Text] [PDF]

				Y. A. Elsayed and E. A. Sausville Selected Novel Anticancer Treatments Targeting Cell Signaling Proteins Oncologist, December 1, 2001; 6(6): 517 - 537. [Abstract] [Full Text] [PDF]

				Y. Dai, C. Yu, V. Singh, L. Tang, Z. Wang, R. McInistry, P. Dent, and S. Grant Pharmacological Inhibitors of the Mitogen-activated Protein Kinase (MAPK) Kinase/MAPK Cascade Interact Synergistically with UCN-01 to Induce Mitochondrial Dysfunction and Apoptosis in Human Leukemia Cells Cancer Res., July 1, 2001; 61(13): 5106 - 5115. [Abstract] [Full Text] [PDF]

				E. A. Sausville, S. G. Arbuck, R. Messmann, D. Headlee, K. S. Bauer, R. M. Lush, A. Murgo, W. D. Figg, T. Lahusen, S. Jaken, et al. Phase I Trial of 72-Hour Continuous Infusion UCN-01 in Patients With Refractory Neoplasms J. Clin. Oncol., April 15, 2001; 19(8): 2319 - 2333. [Abstract] [Full Text] [PDF]

				S. Harvey, R. Decker, Y. Dai, G. Schaefer, L. Tang, L. Kramer, P. Dent, and S. Grant Interactions between 2-Fluoroadenine 9-{beta}-D-Arabinofuranoside and the Kinase Inhibitor UCN-01 in Human Leukemia and Lymphoma Cells Clin. Cancer Res., February 1, 2001; 7(2): 320 - 330. [Abstract] [Full Text]

				J. Lozano, S. Menendez, A. Morales, D. Ehleiter, W.-C. Liao, R. Wagman, A. Haimovitz-Friedman, Z. Fuks, and R. Kolesnick Cell Autonomous Apoptosis Defects in Acid Sphingomyelinase Knockout Fibroblasts J. Biol. Chem., January 5, 2001; 276(1): 442 - 448. [Abstract] [Full Text] [PDF]

				E. C. Busby, D. F. Leistritz, R. T. Abraham, L. M. Karnitz, and J. N. Sarkaria The Radiosensitizing Agent 7-Hydroxystaurosporine (UCN-01) Inhibits the DNA Damage Checkpoint Kinase hChk1 Cancer Res., April 1, 2000; 60(8): 2108 - 2112. [Abstract] [Full Text]

				W. H. Wilson, L. Sorbara, W. D. Figg, E. K. Mont, E. Sausville, K. E. Warren, F. M. Balis, K. Bauer, M. Raffeld, A. M. Senderowicz, et al. Modulation of Clinical Drug Resistance in a B Cell Lymphoma Patient by the Protein Kinase Inhibitor 7-Hydroxystaurosporine: Presentation of a Novel Therapeutic Paradigm Clin. Cancer Res., February 1, 2000; 6(2): 415 - 421. [Abstract] [Full Text]

				K. Sugiyama, T. Akiyama, M. Shimizu, T. Tamaoki, C. Courage, A. Gescher, and S. Akinaga Decrease in Susceptibility Toward Induction of Apoptosis and Alteration in G1 Checkpoint Function as Determinants of Resistance of Human Lung Cancer Cells against the Antisignaling Drug UCN-01 (7-Hydroxystaurosporine) Cancer Res., September 1, 1999; 59(17): 4406 - 4412. [Abstract] [Full Text] [PDF]

				P. C. Mack, D. R. Gandara, C. Bowen, M. J. Edelman, T. Paglieroni, J. B. Schnier, E. P. Gelmann, and P. H. Gumerlock RB Status as a Determinant of Response to UCN-01 in Non-Small Cell Lung Carcinoma Clin. Cancer Res., September 1, 1999; 5(9): 2596 - 2604. [Abstract] [Full Text] [PDF]

				L. Yu, L. Orlandi, P. Wang, M. S. Orr, A. M. Senderowicz, E. A. Sausville, R. Silvestrini, N. Watanabe, H. Piwnica-Worms, and P. M. OConnor UCN-01 Abrogates G2 Arrest through a Cdc2-dependent Pathway That Is Associated with Inactivation of the Wee1Hu Kinase and Activation of the Cdc25C Phosphatase J. Biol. Chem., December 11, 1998; 273(50): 33455 - 33464. [Abstract] [Full Text] [PDF]

				S. T. Harkin, G. M. Cohen, and A. Gescher Modulation of Apoptosis in Rat Thymocytes by Analogs of Staurosporine: Lack of Direct Association with Inhibition of Protein Kinase C Mol. Pharmacol., October 1, 1998; 54(4): 663 - 670. [Abstract] [Full Text] [PDF]

				B. W. Parker, G. Kaur, W. Nieves-Neira, M. Taimi, G. Kohlhagen, T. Shimizu, M. D. Losiewicz, Y. Pommier, E. A. Sausville, and A. M. Senderowicz Early Induction of Apoptosis in Hematopoietic Cell Lines After Exposure to Flavopiridol Blood, January 15, 1998; 91(2): 458 - 465. [Abstract] [Full Text] [PDF]

				R.-G. Shao, C.-X. Cao, and Y. Pommier Activation of PKC{alpha} Downstream from Caspases during Apoptosis Induced by 7-Hydroxystaurosporine or the Topoisomerase Inhibitors, Camptothecin and Etoposide, in Human Myeloid Leukemia HL60 Cells J. Biol. Chem., December 12, 1997; 272(50): 31321 - 31325. [Abstract] [Full Text] [PDF]

				S. Wang, J. A. Vrana, T. M. Bartimole, A. J. Freemerman, W. D. Jarvis, L. B. Kramer, G. Krystal, P. Dent, and S. Grant Agents that Down-Regulate or Inhibit Protein Kinase C Circumvent Resistance to 1-{beta}-D-Arabinofuranosylcytosine-Induced Apoptosis in Human Leukemia Cells that Overexpress Bcl-2 Mol. Pharmacol., December 1, 1997; 52(6): 1000 - 1009. [Abstract] [Full Text] [PDF]

				J. Lozano, S. Menendez, A. Morales, D. Ehleiter, W.-C. Liao, R. Wagman, A. Haimovitz-Friedman, Z. Fuks, and R. Kolesnick Cell Autonomous Apoptosis Defects in Acid Sphingomyelinase Knockout Fibroblasts J. Biol. Chem., January 5, 2001; 276(1): 442 - 448. [Abstract] [Full Text] [PDF]