This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Furuya, Y. Articles by Isaacs, J. T. Search for Related Content PubMed PubMed Citation Articles by Furuya, Y. Articles by Isaacs, J. T.

The Role of Calcium, pH, and Cell Proliferation in the Programmed (Apoptotic) Death of Androgen-independent Prostatic Cancer Cells Induced by Thapsigargin¹

The Johns Hopkins Oncology Center [Y. F., P. L., J. T. I.], James Buchana Brady Urological Institute and the Department of Urology [J. T. I.], Johns Hopkins School of Medicine, Baltimore, Maryland 21205, and The Department of Biochemistry, University of Maryland School of Medicine, Baltimore, Maryland 21201 [A. D. S., D. L. G.]

Calcium (Ca²⁺) accumulates within the endoplasmic reticulum of cells through function of the sarcoplasmic reticulum and endoplasmic reticulum Ca²⁺-dependent ATPase family of intracellular Ca²⁺-pumping ATPases. The resulting pools have important signaling functions. Thapsigargin (TG) is a sesquiterpene -lactone which selectively inhibits the sarcoplasmic reticulum and endoplasmic reticulum Ca²⁺-dependent ATPase pumps with a 50% inhibitory concentration of approximately 30 nM. Treatment of androgen-independent prostate cancer cells of both rat and human origin with TG inhibits their endoplasmic reticulum Ca²⁺-dependent ATPase activity, resulting in a 3–4-fold elevation in the level of intracellular free Ca²⁺ (Ca_i) within minutes of exposure. Due to a secondary influx of extracellular Ca²⁺, this increase in Ca_i is sustained, resulting in morphological (cell rounding) and biochemical changes within 6–12 h (enhanced calmodulin, glucose regulated protein, and tissue transglutaminase expression, and decreased expression of the G₁ cyclins). Within 24 h of exposure, androgen-independent prostatic cancer cells stop progression through the cell cycle, arrest out of cycle in G₀, and irreversibly lose their ability to proliferate with a median effective concentration value of 31 nM TG. During the next 24–48 h, the genomic DNA of the G₀-arrested cells undergoes double-strand fragmentation. This is followed by the loss of plasma membrane integrity and fragmentation of the cell into apoptotic bodies. During this process, there is no acidification in the intracellular pH. Using cells transfected with the avian M_r 28,000 calbindin D Ca²⁺-buffering protein, it was demonstrated that the programmed death initiated by TG is critically dependent upon an adequate (i.e., 3–4-fold) sustained (>1 h) elevation in Ca_i and not depletion of the endoplasmic reticulum pools of Ca²⁺. These results demonstrate that TG induces programmed cell death in androgen-independent prostatic cancer cells in a dose-dependent manner and that this death does not require proliferation or intracellular acidification but is critically dependent upon an adequate, sustained (i.e., >1 h) elevation in Ca_i.

¹ Supported by NIH grant CA58236.

² Present address: Section of Urology, Department of Surgery, University of Trondheim, N-7000 Trondheim, Norway.

³ To whom requests for reprints should be addressed, at Johns Hopkins Oncology Center, 422 N. Bond Street, Baltimore, MD 21231.

Received 4/22/94. Accepted 9/28/94.

This article has been cited by other articles:

				D. J. Vander Griend, L. Antony, S. L. Dalrymple, Y. Xu, S. B. Christensen, S. R. Denmeade, and J. T. Isaacs Amino acid containing thapsigargin analogues deplete androgen receptor protein via synthesis inhibition and induce the death of prostate cancer cells Mol. Cancer Ther., May 1, 2009; 8(5): 1340 - 1349. [Abstract] [Full Text] [PDF]

				M. Garcia, X.-F. Yu, D. E. Griffin, and W. J. Moss Measles virus inhibits human immunodeficiency virus type 1 reverse transcription and replication by blocking cell-cycle progression of CD4+ T lymphocytes J. Gen. Virol., April 1, 2008; 89(4): 984 - 993. [Abstract] [Full Text] [PDF]

				S Humez, M Monet, G Legrand, G Lepage, P Delcourt, and N Prevarskaya Epidermal growth factor-induced neuroendocrine differentiation and apoptotic resistance of androgen-independent human prostate cancer cells. Endocr. Relat. Cancer, March 1, 2006; 13(1): 181 - 195. [Abstract] [Full Text] [PDF]

				H. Yamaguchi and H.-G. Wang Tissue Transglutaminase Serves as an Inhibitor of Apoptosis by Cross-Linking Caspase 3 in Thapsigargin-Treated Cells Mol. Cell. Biol., January 15, 2006; 26(2): 569 - 579. [Abstract] [Full Text] [PDF]

				M. Rota, A. Boni, K. Urbanek, M. E. Padin-Iruegas, T. J. Kajstura, G. Fiore, H. Kubo, E. H. Sonnenblick, E. Musso, S. R. Houser, et al. Nuclear Targeting of Akt Enhances Ventricular Function and Myocyte Contractility Circ. Res., December 9, 2005; 97(12): 1332 - 1341. [Abstract] [Full Text] [PDF]

				C. H. Park, E. R. Hahm, J. H. Lee, K. C. Jung, H. S. Rhee, and C. H. Yang Ionomycin downregulates {beta}-catenin/Tcf signaling in colon cancer cell line Carcinogenesis, November 1, 2005; 26(11): 1929 - 1933. [Abstract] [Full Text] [PDF]

				A. B. Parekh and J. W. Putney Jr. Store-Operated Calcium Channels Physiol Rev, April 1, 2005; 85(2): 757 - 810. [Abstract] [Full Text] [PDF]

				L. Zhang and G. J. Barritt Evidence that TRPM8 Is an Androgen-Dependent Ca2+ Channel Required for the Survival of Prostate Cancer Cells Cancer Res., November 15, 2004; 64(22): 8365 - 8373. [Abstract] [Full Text] [PDF]

				S. Janssen, C. M. Jakobsen, D. M. Rosen, R. M. Ricklis, U. Reineke, S. B. Christensen, H. Lilja, and S. R. Denmeade Screening a combinatorial peptide library to develop a human glandular kallikrein 2-activated prodrug as targeted therapy for prostate cancer Mol. Cancer Ther., November 1, 2004; 3(11): 1439 - 1450. [Abstract] [Full Text] [PDF]

				S. V. Ley, A. Antonello, E. P. Balskus, D. T. Booth, S. B. Christensen, E. Cleator, H. Gold, K. Hogenauer, U. Hunger, R. M. Myers, et al. Natural Product Synthesis Special Feature: Synthesis of the thapsigargins PNAS, August 17, 2004; 101(33): 12073 - 12078. [Abstract] [Full Text] [PDF]

				T. Milakovic, J. Tucholski, E. McCoy, and G. V. W. Johnson Intracellular Localization and Activity State of Tissue Transglutaminase Differentially Impacts Cell Death J. Biol. Chem., March 5, 2004; 279(10): 8715 - 8722. [Abstract] [Full Text] [PDF]

				E. Cifuentes, J. M. Mataraza, B. A. Yoshida, M. Menon, D. B. Sacks, E. R. Barrack, and G. P.-V. Reddy Physical and functional interaction of androgen receptor with calmodulin in prostate cancer cells PNAS, January 13, 2004; 101(2): 464 - 469. [Abstract] [Full Text] [PDF]

				S. R. Denmeade, C. M. Jakobsen, S. Janssen, S. R. Khan, E. S. Garrett, H. Lilja, S. B. Christensen, and J. T. Isaacs Prostate-Specific Antigen-Activated Thapsigargin Prodrug as Targeted Therapy for Prostate Cancer J Natl Cancer Inst, July 2, 2003; 95(13): 990 - 1000. [Abstract] [Full Text] [PDF]

				H. Yamaguchi, K. Bhalla, and H.-G. Wang Bax Plays a Pivotal Role in Thapsigargin-induced Apoptosis of Human Colon Cancer HCT116 Cells by Controlling Smac/Diablo and Omi/HtrA2 Release from Mitochondria Cancer Res., April 1, 2003; 63(7): 1483 - 1489. [Abstract] [Full Text] [PDF]

				P. H. Johnson, R. P. Walker, S. W. Jones, K. Stephens, J. Meurer, D. A. Zajchowski, M. M. Luke, F. Eeckman, Y. Tan, L. Wong, et al. Multiplex Gene Expression Analysis for High-Throughput Drug Discovery: Screening and Analysis of Compounds Affecting Genes Overexpressed in Cancer Cells Mol. Cancer Ther., December 1, 2002; 1(14): 1293 - 1304. [Abstract] [Full Text] [PDF]

				L. Song, P. De Sarno, and R. S. Jope Central Role of Glycogen Synthase Kinase-3beta in Endoplasmic Reticulum Stress-induced Caspase-3 Activation J. Biol. Chem., November 15, 2002; 277(47): 44701 - 44708. [Abstract] [Full Text] [PDF]

				S. Gil-Parrado, A. Fernandez-Montalvan, I. Assfalg-Machleidt, O. Popp, F. Bestvater, A. Holloschi, T. A. Knoch, E. A. Auerswald, K. Welsh, J. C. Reed, et al. Ionomycin-activated Calpain Triggers Apoptosis. A PROBABLE ROLE FOR Bcl-2 FAMILY MEMBERS J. Biol. Chem., July 19, 2002; 277(30): 27217 - 27226. [Abstract] [Full Text] [PDF]

				L. K. Nutt, J. Chandra, A. Pataer, B. Fang, J. A. Roth, S. G. Swisher, R. G. O'Neil, and D. J. McConkey Bax-mediated Ca2+ Mobilization Promotes Cytochrome c Release during Apoptosis J. Biol. Chem., May 31, 2002; 277(23): 20301 - 20308. [Abstract] [Full Text] [PDF]

				L. K. Nutt, A. Pataer, J. Pahler, B. Fang, J. Roth, D. J. McConkey, and S. G. Swisher Bax and Bak Promote Apoptosis by Modulating Endoplasmic Reticular and Mitochondrial Ca2+ Stores J. Biol. Chem., March 8, 2002; 277(11): 9219 - 9225. [Abstract] [Full Text] [PDF]

				G. Legrand, S. Humez, C. Slomianny, E. Dewailly, F. V. Abeele, P. Mariot, F. Wuytack, and N. Prevarskaya Ca2+ Pools and Cell Growth. EVIDENCE FOR SARCOENDOPLASMIC Ca2+-ATPases 2B INVOLVEMENT IN HUMAN PROSTATE CANCER CELL GROWTH CONTROL J. Biol. Chem., December 7, 2001; 276(50): 47608 - 47614. [Abstract] [Full Text] [PDF]

				M. Unkila, K. S. McColl, M. J. Thomenius, K. Heiskanen, and C. W. Distelhorst Unreliability of the Cytochrome c-Enhanced Green Fluorescent Fusion Protein as a Marker of Cytochrome c Release in Cells That Overexpress Bcl-2 J. Biol. Chem., October 12, 2001; 276(42): 39132 - 39137. [Abstract] [Full Text] [PDF]

				J. Pinski, A. Parikh, G. S. Bova, and J. T. Isaacs Therapeutic Implications of Enhanced G0/G1 Checkpoint Control Induced by Coculture of Prostate Cancer Cells with Osteoblasts Cancer Res., September 1, 2001; 61(17): 6372 - 6376. [Abstract] [Full Text] [PDF]

				M. Chami, D. Gozuacik, D. Lagorce, M. Brini, P. Falson, G. Peaucellier, P. Pinton, H. Lecoeur, M.-L. Gougeon, M. le Maire, et al. Serca1 Truncated Proteins Unable to Pump Calcium Reduce the Endoplasmic Reticulum Calcium Concentration and Induce Apoptosis J. Cell Biol., June 11, 2001; 153(6): 1301 - 1314. [Abstract] [Full Text] [PDF]

				R. Skryma, P. Mariot, X. Le Bourhis, F. Van Coppenolle, Y. Shuba, F. V. Abeele, G. Legrand, S. Humez, B. Boilly, and N. Prevarskaya Store depletion and store-operated Ca2+ current in human prostate cancer LNCaP cells: involvement in apoptosis J. Physiol., August 15, 2000; 527(1): 71 - 83. [Abstract] [Full Text] [PDF]

				D. J. McConkey, Y. Lin, L. K. Nutt, H. Z. Ozel, and R. A. Newman Cardiac Glycosides Stimulate Ca2+ Increases and Apoptosis in Androgen-independent, Metastatic Human Prostate Adenocarcinoma Cells Cancer Res., July 1, 2000; 60(14): 3807 - 3812. [Abstract] [Full Text]

				C. Jackisch, H. A. Hahm, B. Tombal, D. McCloskey, K. Butash, N. E. Davidson, and S. R. Denmeade Delayed Micromolar Elevation in Intracellular Calcium Precedes Induction of Apoptosis in Thapsigargin-treated Breast Cancer Cells Clin. Cancer Res., July 1, 2000; 6(7): 2844 - 2850. [Abstract] [Full Text]

				G. Barnett, A.-M. Jakobsen, M. Tas, K. Rice, J. Carmichael, and J. C. Murray Prostate Adenocarcinoma Cells Release the Novel Proinflammatory Polypeptide EMAP-II in Response to Stress Cancer Res., June 1, 2000; 60(11): 2850 - 2857. [Abstract] [Full Text] [PDF]

				I. E. Wertz and V. M. Dixit Characterization of Calcium Release-activated Apoptosis of LNCaP Prostate Cancer Cells J. Biol. Chem., April 6, 2000; 275(15): 11470 - 11477. [Abstract] [Full Text] [PDF]

				P. R. Turner, S. Mefford, S. Christakos, and R. A. Nissenson Apoptosis Mediated by Activation of the G Protein-Coupled Receptor for Parathyroid Hormone (PTH)/ PTH-Related Protein (PTHrP) Mol. Endocrinol., February 1, 2000; 14(2): 241 - 254. [Abstract] [Full Text]

				D. M. Haverstick, T. N. Heady, T. L. Macdonald, and L. S. Gray Inhibition of Human Prostate Cancer Proliferation in Vitro and in a Mouse Model by a Compound Synthesized to Block Ca2+ Entry Cancer Res., February 1, 2000; 60(4): 1002 - 1008. [Abstract] [Full Text]

				L. Zhu, S. Ling, X.-D. Yu, L. K. Venkatesh, T. Subramanian, G. Chinnadurai, and T. H. Kuo Modulation of Mitochondrial Ca2+ Homeostasis by Bcl-2 J. Biol. Chem., November 19, 1999; 274(47): 33267 - 33273. [Abstract] [Full Text] [PDF]

				A. A Gutierrez, J. M. Arias, L. Garcia, J. Mas-Oliva, and A. Guerrero-Hernandez Activation of a Ca2+-permeable cation channel by two different inducers of apoptosis in a human prostatic cancer cell line J. Physiol., May 15, 1999; 517(1): 95 - 107. [Abstract] [Full Text] [PDF]

				H. Wang, N. Pathan, I. M. Ethell, S. Krajewski, Y. Yamaguchi, F. Shibasaki, F. McKeon, T. Bobo, T. F. Franke, and J. C. Reed Ca2+-Induced Apoptosis Through Calcineurin Dephosphorylation of BAD Science, April 9, 1999; 284(5412): 339 - 343. [Abstract] [Full Text]

				N. Zhu and Z. Wang Calreticulin Expression Is Associated with Androgen Regulation of the Sensitivity to Calcium Ionophore-induced Apoptosis in LNCaP Prostate Cancer Cells Cancer Res., April 1, 1999; 59(8): 1896 - 1902. [Abstract] [Full Text] [PDF]

				Y.-M. Fu, Z.-X. Yu, B. A. Pelayo, V. J. Ferrans, and G. G. Meadows Focal Adhesion Kinase-dependent Apoptosis of Melanoma Induced by Tyrosine and Phenylalanine Deficiency Cancer Res., February 1, 1999; 59(3): 758 - 765. [Abstract] [Full Text] [PDF]

				R. Benson, C Dive, and A. Watson Cytoplasmic acidification is not an effector mechanism of VP16 or DEX-induced apoptosis in CEM T leukaemia cells J. Cell Sci., January 6, 1999; 112(11): 1755 - 1760. [Abstract] [PDF]

				J. A. Maloney, O. M. Tsygankova, L. Yang, Q. Li, A. Szot, K. Baysal, and J. R. Williamson Activation of ERK by Ca2+ store depletion in rat liver epithelial cells Am J Physiol Cell Physiol, January 1, 1999; 276(1): C221 - C230. [Abstract] [Full Text] [PDF]

				R.S. Fife and G.W. Sledge JR Effects of Doxycycline on Cancer Cells in Vitro and in Vivo Advances in Dental Research, November 1, 1998; 12(1): 94 - 96. [PDF]

				H. He, M. Lam, T. S. McCormick, and C. W. Distelhorst Maintenance of Calcium Homeostasis in the Endoplasmic Reticulum by Bcl-2 J. Cell Biol., September 22, 1997; 138(6): 1219 - 1228. [Abstract] [Full Text] [PDF]

				P. Nair, S. Muthukkumar, S. F. Sells, S.-S. Han, V. P. Sukhatme, and V. M. Rangnekar Early Growth Response-1-dependent Apoptosis Is Mediated by p53 J. Biol. Chem., August 8, 1997; 272(32): 20131 - 20138. [Abstract] [Full Text] [PDF]

				R. T. Waldron, A. D. Short, and D. L. Gill Store-operated Ca2+ Entry and Coupling to Ca2+ Pool Depletion in Thapsigargin-resistant Cells J. Biol. Chem., March 7, 1997; 272(10): 6440 - 6447. [Abstract] [Full Text] [PDF]

				C. A. Ufret-Vincenty, A. D. Short, A. Alfonso, and D. L. Gill A Novel Ca[IMAGE] Entry Mechanism Is Turned On during Growth Arrest Induced by Ca[IMAGE] Pool Depletion J. Biol. Chem., November 10, 1995; 270(45): 26790 - 26793. [Abstract] [Full Text] [PDF]

				R. T. Waldron, A. D. Short, and D. L. Gill Thapsigargin-resistant Intracellular Calcium Pumps J. Biol. Chem., May 19, 1995; 270(20): 11955 - 11961. [Abstract] [Full Text] [PDF]