| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Biochemistry |
Department of Biochemistry and Molecular Biology, Wright State University School of Medicine, Dayton, Ohio 45435
We have assessed in detail the effect of cisplatin-activated programmed cell death in the cisplatin-sensitive human ovarian cancer cell line A2780 and two drug-resistant subclones, CP70 and C30. To determine whether the differential extent of apoptosis observed between the sensitive and resistant ovarian cancer cell lines was the result of dissimilar upstream signaling events, we assessed the execution of apoptotic events that precede target protein proteolysis and subsequent chromosomal DNA degradation. Proteolytic degradation of procaspase-3 was observed in both the CP70 and C30 cells following IC50 cisplatin treatment, whereas no proteolyzed caspase-3 subunits were detected in the A2780 cells. However, using a direct enzymatic assay measuring cleavage of the synthetic peptide substrate (N-acetyl-Asp-Glu-Val-Asp-p-nitroanilide), activity was detected in extracts prepared from A2780 cells treated at the IC90 level of cisplatin and was 2–3-fold less than that of extracts prepared from CP70 and C30 cells. Because the activation of procaspase-3 by caspase-9 requires the release of cytochrome c into the cytoplasm, we determined the level of cytoplasmic cytochrome c in each cell line in response to cisplatin treatment. Consistent with the caspase-3 activation data, a very small increase in cytoplasmic cytochrome c was observed in A2780 cells following cisplatin treatment, whereas dramatic increases were evident in both the CP70 and C30 cell lines. The expression of the mitochondrial factors Bcl-2, Bcl-x, and Bax was determined because each has been implicated in the regulation or release of cytochrome c at the level of the mitochondria. Bcl-2 and Bcl-xL proteins remained relatively unchanged in expression for over 48 h after exposure to cisplatin in the A2780 cell lines. However, within the same time period, expression of Bcl-2 decreased in the CP70- and C30-resistant cell lines, whereas an increase in Bcl-xL expression was observed. Expression of the proapoptotic Bcl-xS protein was observed in only the resistant CP70 and C30 cell lines independent of cisplatin treatment. A change in the expression of Mr 24,000 Bax to a Mr 21,000 isoform was evidenced in the A2780 cells within 48 h of cisplatin treatment and, to a greater extent, in the CP70 and C30 cells, which also expressed a Mr 16,000 Bax variant. Evidence for an alternative apoptotic pathway in A2780 cells was obtained by demonstrating increased FADD expression in response to cisplatin treatment. These results support a model in which cisplatin-induced programmed cell death in the cisplatin-sensitive A2780 and -resistant CP70 and C30 cells proceeds via caspase-3-independent and -dependent pathways, respectively.
This article has been cited by other articles:
|
|
 |
|
  M. Muscolini, R. Cianfrocca, A. Sajeva, S. Mozzetti, G. Ferrandina, A. Costanzo, and L. Tuosto Trichostatin A up-regulates p73 and induces Bax-dependent apoptosis in cisplatin-resistant ovarian cancer cells Mol. Cancer Ther., June 1, 2008; 7(6): 1410 - 1419. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Zhong, Y.-J. Wen, H.-S. Yang, H. Luo, A.-F. Fu, F. Yang, L.-J. Chen, X. Chen, X.-R. Qi, H.-G. Lin, et al. Efficient inhibition of cisplatin-resistant human ovarian cancer growth and prolonged survival by gene transferred vesicular stomatitis virus matrix protein in nude mice Ann. Onc., April 23, 2008; (2008) mdn167v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. G. M. Cillessen, J. C. Reed, K. Welsh, C. Pinilla, R. Houghten, E. Hooijberg, J. Deurhof, K. C. M. Castricum, P. Kortman, C. J. Hess, et al. Small-molecule XIAP antagonist restores caspase-9 mediated apoptosis in XIAP-positive diffuse large B-cell lymphoma cells Blood, January 1, 2008; 111(1): 369 - 375. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A.G.M. Cillessen, C. J. Hess, E. Hooijberg, K. C.M. Castricum, P. Kortman, F. Denkers, W. Vos, M. A. van de Wiel, G. J. Schuurhuis, G. J. Ossenkoppele, et al. Inhibition of the Intrinsic Apoptosis Pathway Downstream of Caspase-9 Activation Causes Chemotherapy Resistance in Diffuse Large B-Cell Lymphoma Clin. Cancer Res., December 1, 2007; 13(23): 7012 - 7021. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Razzaque Cisplatin nephropathy: is cytotoxicity avoidable? Nephrol. Dial. Transplant., August 1, 2007; 22(8): 2112 - 2116. [Full Text] [PDF]
|
|
|
|
|
|
Y. Zhang and X. Shen Heat Shock Protein 27 Protects L929 Cells from Cisplatin-Induced Apoptosis by Enhancing Akt Activation and Abating Suppression of Thioredoxin Reductase Activity Clin. Cancer Res., May 15, 2007; 13(10): 2855 - 2864. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Tarang, A. Sodhi, and P. Chauhan Differential expression of Toll-like receptors in murine peritoneal macrophages in vitro on treatment with cisplatin Int. Immunol., May 1, 2007; 19(5): 635 - 643. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Lebrun, M. R. Montminy, and E. Van Obberghen Regulation of the Pancreatic Duodenal Homeobox-1 Protein by DNA-dependent Protein Kinase J. Biol. Chem., November 18, 2005; 280(46): 38203 - 38210. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-C. YEN, Y.-C. TANG, F.-Y. CHEN, S.-W. CHEN, and H. J. MAJIMA Enhancement of Cisplatin-Induced Apoptosis and Caspase 3 Activation by Depletion of Mitochondrial DNA in a Human Osteosarcoma Cell Line Ann. N.Y. Acad. Sci., May 1, 2005; 1042(1): 516 - 522. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. J. Boeckman, K. S. Trego, and J. J. Turchi Cisplatin Sensitizes Cancer Cells to Ionizing Radiation via Inhibition of Nonhomologous End Joining Mol. Cancer Res., May 1, 2005; 3(5): 277 - 285. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Anderson, K. A. Lawson, M. Simmons-Menchaca, L. Sun, B. G. Sanders, and K. Kline {alpha}-TEA Plus Cisplatin Reduces Human Cisplatin-Resistant Ovarian Cancer Cell Tumor Burden and Metastasis Experimental Biology and Medicine, December 1, 2004; 229(11): 1169 - 1176. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. Pingle, S. Mishra, A. Marcuzzi, S. G. Bhat, Y. Sekino, L. P. Rybak, and V. Ramkumar Osmotic Diuretics Induce Adenosine A1 Receptor Expression and Protect Renal Proximal Tubular Epithelial Cells against Cisplatin-mediated Apoptosis J. Biol. Chem., October 8, 2004; 279(41): 43157 - 43167. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. C. Han, M.-Y. Lee, K. D. Shin, S. B. Jeon, J. M. Kim, K.-H. Son, H.-C. Kim, H.-M. Kim, and B.-M. Kwon 2'-Benzoyloxycinnamaldehyde Induces Apoptosis in Human Carcinoma via Reactive Oxygen Species J. Biol. Chem., February 20, 2004; 279(8): 6911 - 6920. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Anderson, W. Alrefai, P. Bonomi, T. M. Seed, P. Dudeja, Y. Hu, and J. E. Harris Caspase-Dependent and -Independent Panc-1 Cell Death Due to Actinomycin D and MK 886 Are Additive but Increase Clonogenic Survival Experimental Biology and Medicine, September 1, 2003; 228(8): 915 - 925. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Nowak Protein Kinase C-alpha and ERK1/2 Mediate Mitochondrial Dysfunction, Decreases in Active Na+ Transport, and Cisplatin-induced Apoptosis in Renal Cells J. Biol. Chem., November 1, 2002; 277(45): 43377 - 43388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. E. Chalfant, K. Rathman, R. L. Pinkerman, R. E. Wood, L. M. Obeid, B. Ogretmen, and Y. A. Hannun De Novo Ceramide Regulates the Alternative Splicing of Caspase 9 and Bcl-x in A549 Lung Adenocarcinoma Cells. DEPENDENCE ON PROTEIN PHOSPHATASE-1 J. Biol. Chem., April 5, 2002; 277(15): 12587 - 12595. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Huang, R. T. Dunn II, S. Jayadev, O. DiSorbo, F. D. Pack, S. B. Farr, R. E. Stoll, and K. T. Blanchard Assessment of Cisplatin-Induced Nephrotoxicity by Microarray Technology Toxicol. Sci., October 1, 2001; 63(2): 196 - 207. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. M. Gonzalez, M. A. Fuertes, C. Alonso, and J. M. Perez Is Cisplatin-Induced Cell Death Always Produced by Apoptosis? Mol. Pharmacol., April 1, 2001; 59(4): 657 - 663. [Full Text]
|
|
|
|
 |
|
 R. N. Winter, A. Kramer, A. Borkowski, and N. Kyprianou Loss of Caspase-1 and Caspase-3 Protein Expression in Human Prostate Cancer Cancer Res., February 1, 2001; 61(3): 1227 - 1232. [Abstract] [Full Text]
|
|
|
|
|
|
Y. Mitsuuchi, S. W. Johnson, M. Selvakumaran, S. J. Williams, T. C. Hamilton, and J. R. Testa The Phosphatidylinositol 3-Kinase/AKT Signal Transduction Pathway Plays a Critical Role in the Expression of p21WAF1/CIP1/SDI1 Induced by Cisplatin and Paclitaxel Cancer Res., October 1, 2000; 60(19): 5390 - 5394. [Abstract] [Full Text]
|
|
|
|
 |
|
 P. Costantini, E. Jacotot, D. Decaudin, and G. Kroemer Mitochondrion as a Novel Target of Anticancer Chemotherapy J Natl Cancer Inst, July 5, 2000; 92(13): 1042 - 1053. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Schuler, E. Bossy-Wetzel, J. C. Goldstein, P. Fitzgerald, and D. R. Green p53 Induces Apoptosis by Caspase Activation through Mitochondrial Cytochrome c Release J. Biol. Chem., March 15, 2000; 275(10): 7337 - 7342. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Fujii, M. L. Garcia-Bermejo, J. L. Bernabo, J. Caamano, M. Ohba, T. Kuroki, L. Li, S. H. Yuspa, and M. G. Kazanietz Involvement of Protein Kinase C delta (PKCdelta ) in Phorbol Ester-induced Apoptosis in LNCaP Prostate Cancer Cells. LACK OF PROTEOLYTIC CLEAVAGE OF PKCdelta J. Biol. Chem., March 10, 2000; 275(11): 7574 - 7582. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Cancer Research | Clinical Cancer Research |
| Cancer Epidemiology Biomarkers & Prevention | Molecular Cancer Therapeutics |
| Molecular Cancer Research | Cancer Prevention Research |
| Cancer Prevention Journals Portal | Cancer Reviews Online |
| Annual Meeting Education Book | Cell Growth & Differentiation |
