Cancer Research Cancer Research Funding Available Protein Translation and Cancer
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 Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Maestre, N.
Right arrow Articles by Jaffrézou, J.-P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Maestre, N.
Right arrow Articles by Jaffrézou, J.-P.
[Cancer Research 61, 2558-2561, March 15, 2001]
© 2001 American Association for Cancer Research

Experimental Therapeutics

Cell Surface-directed Interaction of Anthracyclines Leads to Cytotoxicity and Nuclear Factor {kappa}B Activation but not Apoptosis Signaling1

Nicolas Maestre, Thomas R. Tritton, Guy Laurent and Jean-Pierre Jaffrézou2

Institut Nationale de la Santé et de la Recherche Médicale E9910, Institut Claudius Régaud, 31052 Toulouse, France [N. M., G. L., J-P. J.]; Haverford College, Haverford, Pennsylvania 19041 [T. R. T.]; and the Service d’Hématologie, Centre Hospitalier Universitaire Purpan, 31059 Toulouse, France [G. L.]


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
Anthracyclines are, above all, DNA intercalators, which induce genetic damage leading to cell death. However, increasing evidence firmly suggests that the underlying mechanism for anthracycline cytotoxicity is the induction of apoptosis through intracellular-mediated signaling pathways. Whether drug/DNA interaction is necessary for such apoptosis signaling is unknown. We investigated the cellular effects of the anthracyclines daunorubicin (DNR) and doxorubicin (DOX) using the myeloid leukemia cell line U937. By comparing free drug against agarose bead-immobilized drug iDNR and iDOX (which cannot accumulate within the cell), we observed that whereas both free and immobilized anthracyclines were cytotoxic, only the former induced apoptosis; the latter induced necrosis. Indeed, we did not observe ceramide generation, neutral sphingomyelinase activation, poly (ADP-ribose) polymerase cleavage, or other apoptotic events with iDNR or iDOX. However, both free and immobilized drug were similarly capable of triggering nuclear factor {kappa}B activation. These observations demonstrate that whereas activation of certain cellular signaling pathways can be achieved solely through membrane interaction, apoptosis signaling requires anthracycline internalization. These results also show that the initiation of cell survival pathways (illustrated by nuclear factor {kappa}B activation) is independent of intracellular drug/target interaction.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 
Anthracyclines are one of the most active antitumor compounds used in clinical oncology, especially in the treatment of acute leukemias. Anthracycline cytotoxicity is generally believed to be the result of drug-induced DNA damage. Such damage can result from the intercalation-induced distortion of the double-helix or by stabilization of the cleavable complex formed between DNA and topoisomerase II (reviewed in Ref. 1 ). However, how and why such events should bring about cell death remains unclear, especially when one considers that DNA interaction may not be a prerequisite for anthracycline cytotoxicity (2, 3, 4, 5) . Hence, the exploration and understanding of the process of apoptosis has forced a reconsideration of the mechanisms whereby cells respond to anthracyclines.

We previously demonstrated that DNR3 activates the sphingomyelin-ceramide cycle. Indeed, DNR stimulated the neutral sphingomyelinase activity responsible for sphingomyelin hydrolysis and the subsequent ceramide generation in U937 and HL-60 human leukemia cells (6) . The fact that cell-permeant ceramides, as well as natural ceramide (generated by exposure of the cells to bacterial sphingomyelinase), induce apoptosis in these cells strongly suggests that ceramide was the mediator of DNR-induced apoptosis.

Present knowledge does not allow us to determine whether apoptosis signaling is a consequence of DNR-induced DNA lesions or originates from an independently triggered signaling pathway. To elucidate the origin of apoptosis signaling by anthracyclines, we investigated the cellular effects of the DNR and DOX using the myeloid leukemia cell line U937. By comparing free drug against agarose bead-immobilized drug (iDNR and iDOX), we observed that whereas both free and immobilized anthracyclines were cytotoxic, only the former triggered an apoptotic-signaling pathway. However, the transcription factor NF{kappa}B was similarly activated by both free and immobilized anthracyclines.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS AND DISCUSSION
 REFERENCES
 
Drugs and Reagents.
DNR and DOX was obtained from the National Cancer Institute Drug Repository. Aquasafe 300 scintillation cocktail was purchased from Berthold (Elancourt, France). Silica gel 60 TLC plates were from Merck (Darmstadt, Germany). NF{kappa}B consensus oligonucleotides were obtained from Promega (Madison, WI), and poly(dI-dC) from Pharmacia Biotech (St. Quentin Yvelines, France). All other drugs and reagents were purchased from Sigma, Carlo Erba (Rueil-Malmaison, France), or Prolabo (Paris, France).

Preparation of Agarose Beads-Anthracycline Complexes.
Synthesis of agarose beads-anthracycline complexes was performed as described previously (7) . Briefly, about 1.5 x 10-8 M anthracycline was allowed to react per mg of activated agarose (Reacti-gel 6x, Pierce Chemical; the molar ratio of 1, 1'-carbonyldiimidazole to drug was approximately 12:1). The synthesis is carried out for 48 h in 0.1 M borate buffer (pH 8.0) at 4°C. Under these conditions, ~30% of the drug becomes covalently attached to the beads; the remaining unreacted imidazole carbamate groups are eliminated with an excess of hydroxylamine. Unattached drug is removed by exhaustive washing with borate buffer, acetonitrile, and methanol. The resulting immobilized drug is stored in the dark at 4°C. It does not release free drug for at least several months. In experimental assays, iDNR and iDOX were washed three times with media and resuspended in 1 ml of media. To 5 ml of cells (~1. 5 x 106 cells), 500 µl of immobilized drug suspension was added (giving a final concentration of ~0.3 µM total bound anthracycline).

Cell Culture.
The human leukemia cell line U937 was purchased from the American Type Culture Collection (Rockville, MD). Cells were cultured in RPMI 1640 supplemented with 10% heat-inactivated FCS, 2 mM glutamine, 100 units/ml penicillin and 100 µg/ml streptomycin (all from Eurobio, Les Ulis, France). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO2. Cell stocks were screened routinely for Mycoplasma by the PCR method (Stratagene Mycoplasma PCR kit, La Jolla, CA).

DNA Analyses.
Quantitative DNA fragmentation was determined by the spectrofluorometric DAPI procedure as described previously (8) .

PARP Cleavage.
Analysis of PARP proteolysis was performed by resuspending cells in sample buffer [62.5 mM Tris (pH 6.8) 4 M urea, 10% glycerol, 2% SDS, 5% ß-mercaptoethanol, and 0.04% bromphenol blue). Samples were boiled for 5 min, loaded onto a 10% SDS-polyacrylamide gel. After electrophoresis and transfer onto a nitrocellulose membrane, PARP and its cleaved fragment were detected by using a rabbit polyclonal antiserum (Boehringer-Mannheim, Meylan, France) and a donkey antirabbit secondary antibody (Immunotech, Marseille, France). The signal was visualized by enhanced chemiluminescence (Amersham, Buckinghamshire, United Kingdom).

Metabolic Cell Labeling and Sphingolipid Quantitation.
Sphingomyelin quantitation was performed by labeling cells to isotopic equilibrium with 0.4 µCi/ml of [methyl-[3H]]choline (specific activity 81.0 Ci/mmol; DuPont-NEN, Les Ulis, France) for 48 h in complete medium as described previously (6 , 9) . Then cells were washed and resuspended in serum-free medium for kinetic experiments. Aliquots were taken for protein determination (10) . Radioactive lipids were extracted (11) and sphingomyelin quantitated by scintillation counting (6) .

Total cellular ceramide quantitation was performed by labeling cells to isotopic equilibrium with 1 µCi/ml of] 9, 10-3H[palmitic acid (53.0 Ci/mmol; Amersham) for 48 h in complete medium as described previously (6) . Cells were then washed and resuspended in serum-free medium for kinetic experiments. Lipids were extracted and resolved by TLC, ceramide was scraped and quantitated by liquid scintillation spectrometry. Statistical analyses were performed by Student’s t test.

Sphingomyelinase Assay.
Sphingomyelinase activities were determined as described previously (12) using [choline-methyl-14C]sphingomyelin (54.5 mCi/mmol; DuPont-NEN; 120,000 dpm/assay) as substrate (6) .

Nuclear Extract Preparation.
Extracts were prepared as described previously (13) . Cells (5 x 106) were incubated with or without drugs, after which cells were washed twice with ice-cold PBS and resuspended in 10 mM HEPES (pH 7.8), 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM DTT, 1 mM phenylmethylsulfonyl fluoride, 2 µM pepstatin A, 0.6 µM leupeptin, 1 µg/ml aprotinin, and 0.6% NP40. The nuclei pellet was recovered after centrifugation at 1200 x g and resuspended in 20 mM HEPES (pH 7.9), 0.4 M NaCl, 1 mM EDTA, and 1 mM EGTA. Aliquots were then incubated at 4°C for 30 min, centrifuged at 21,000 x g, and supernatants containing nuclear proteins were removed. Protein concentrations were determined according to Smith et al. (10) using bicinchoninic acid (Sigma, St. Louis, MO).

Electrophoretic Mobility Shift Assays.
Labeling of NF{kappa}B (5'-AGTTGAGGGGACTTTCCCAGGC-3') consensus oligonucleotides (binding sites are underlined) was performed using T4 polynucleotide kinase and [{gamma}32P]ATP (specific activity 5,000 Ci/mmol; Amersham). Binding reactions were carried out in 2 mM HEPES (pH 7.5), 50 mM NaCl, 0.5 mM EDTA, 1 mM MgCl2, 4% glycerol, 0.5 mM DTT, 1 µg poly(dI-dC), and 2 µg of BSA. Typical reactions contained 50,000 cpm of end-labeled NF{kappa}B consensus oligonucleotide (Promega) with 2–6 µg of nuclear extract. After binding (detailed in Figure legends), the mixture was electrophoresed through a low-ionic strength 4% polyacrylamide gel (acrylamide:bisacrylamide ratio, 80:1) containing 6.7 mM Tris-Cl (pH 7.9), 3.3 mM sodium acetate, and 2 mM EDTA. The gel was preelectrophoresed for 90 min at 10 V/cm. Electrophoresis was carried out at the same voltage for 3 h at room temperature with buffer recirculation. The gel was then dried and autoradiographed with intensifying screens at -70°C. Quantification of bands was performed by densitometry and by radioactivity counting of excised bands. Specificity was determined by competition experiments (100-fold excess unlabeled NF{kappa}B or AP-1 consensus oligonucleotide was used) and by super shift assays using p65, p55, and c-Rel specific antibodies generously provided by D. J. Imbert (Institut National de la Santé et de la Recherche Médicale U119, Marseille, France; data not shown).


   RESULTS AND DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
REFERENCES
 
The anthracycline DNR is one of the major antitumor agents widely used in the treatment of AMLs. Cytotoxicity mediated by DNR is generally thought to be the result of drug-induced damage to DNA. However, how and why such an event should bring about cell death remains unclear, especially when one considers that DNA interaction may not be a prerequisite for anthracycline cytotoxicity (2, 3, 4, 5) . Hence, the exploration and understanding of the process of apoptosis has forced a reconsideration of the mechanisms whereby myeloid leukemia cells respond to DNR. In this context, we have shown that, within a narrow concentration range (0.2–1 µM), DNR can trigger apoptosis in U937 or HL-60 AML (8) . These results suggested that DNR triggers apoptotic signals in drug-sensitive AML cells, and that inhibition of these signals may contribute to drug resistance. For this reason, we and others have investigated the mechanism by which DNR activates apoptosis in DNR-sensitive AML cells.

The cytotoxic effect of DNR, DOX, iDNR, and iDOX was evaluated on U937 cells. As expected, both 1 µM DNR and 5 µM DOX (maximal clinically relevant doses) induced significant cytotoxicity, with almost complete elimination of the cellular population within 24 h. iDNR and iDOX were also cytotoxic, inducing an ~20–40% decrease in cell survival within 24 h. Few surviving cells were observed after 72 h (Fig. 1A)Citation . A cytotoxicity profile similar to that observed with the immobilized drug could be obtained by titrating the free drug down to 0.2 µM DNR and 1 µM DOX. Plain (underivatized) agarose beads were without effect. It is important to note that although U937 cells are exposed to 0.3 µM iDNR and iDOX, the actual effective concentration of immobilized anthracycline to which the cells are exposed is far less. Indeed, the "accessible distributed concentration" (i.e., the amount of drug on the surface of the agarose bead) is considered to be <0.01%. However, there is a large focus of drug at the point of contact between the cell and the agarose bead, which probably accounts for the activity of these preparations (2) .



View larger version (19K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 1. Time-dependent cytotoxicity and DNA fragmentation of U937 cells treated with anthracyclines. Cells (3 x 105) were incubated in the absence or in the presence of 1 µM DNR ({circ}), 0.1 µM DNR (•), 5 µM DOX ({square}), 1 µM DOX (), 0.3 µM iDNR (•), 0.3 µM iDOX ({blacksquare}), or plain agarose beads (). At given time points, cell viability was assessed by trypan blue exclusion (A), and DNA fragmentation was determined by the spectrofluorometric DAPI procedure, as described in "Materials and Methods" (B). Results are mean ± SE of three to six independent experiments. **, P < 0.01.

 
To verify whether apoptosis was induced by iDNR and iDOX, we measured DNA fragmentation throughout the cytotoxicity-kinetics experiments. Only DNR and DOX induced significant DNA fragmentation, which correlated well (dose-dependently) with cell death as described previously (Refs. 6 and 8 ; Fig. 1BCitation ). Moreover, morphological analysis of, for example, iDNR-treated U937 cells showed a lack of apoptotic features such as nuclear fragmentation (Fig. 2)Citation . Finally, analysis of PARP cleavage showed that whereas 1 µM DNR and 5 µM DOX cleaved the protein within 6 h (and within 24 h with 0.2 µM DNR and 1 µM DOX), iDNR and iDOX were unable to do so even after 48 h (Fig. 3)Citation .



View larger version (57K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 2. Morphological analysis of U937 cells treated with iDNR. Cells were treated with 0.3 µM iDNR. Morphological alteration of chromatin was evaluated by DAPI staining after 6 h (A), 24 h (B), and 48 h (C) and viewed at a magnification of x20 (A and C) or x100 (B).

 


View larger version (17K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 3. Effect of anthracyclines on PARP cleavage. U937 cells were incubated in the absence (Lane 1) or in the presence of 1 µM DNR (Lane 2) or with 5 µM DOX (Lane 4) for 6 h, or with 0.3 µM iDNR (Lane 5), 0.3 µM iDOX (Lane 3), 0.1 µM DNR (Lane 7), 1 µM DOX (Lane 8), or plain agarose beads (Lane 6) for 48 h. PARP cleavage analysis was then performed as described in "Materials and Methods." Results are representative of at least three independent experiments.

 
To ascertain the potential effects of the anthracyclines on ceramide generation, U937 cells were preincubated with DNR, DOX, iDNR, and iDOX. At several time points, cells were washed and ceramide and sphingomyelin content, as well as neutral sphingomyelinase activity, were measured. As shown in Fig. 4ACitation , and as expected, treatment of U937 cells with DNR and DOX induced a significant burst in intracellular ceramide generation (about 30% to 40% at the highest drug concentration), which was observed concurrently with sphingomyelin hydrolysis at ~6 min (Fig. 4B)Citation . Moreover, both drugs led to an ~30% increase in neutral sphingomyelinase activity, which peaked at 6 min. However, neither plain agarose beads (data not shown), iDNR, nor iDOX presented any significant ceramide generation, sphingomyelin hydrolysis, or sphingomyelinase activation, whereas the equitoxic concentrations of free drug presented an intermediate response (6) , as expected.



View larger version (21K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 4. Effect of anthracyclines on ceramide generation, sphingomyelin hydrolysis, and neutral sphingomyelinase activity in U937 cells. U937 cells were incubated with 1 µM DNR ({circ}), 5 µM DOX ({square}), 0.3 µM iDNR (•), or 0.3 µM iDOX ({blacksquare}) for the time intervals indicated. Ceramide generation (A), sphingomyelin hydrolysis (B), and neutral sphingomyelinase activity (C) was determined as described in "Materials and Methods." Inserts, U937 cells treated with 0.1 µM DNR ({square}) or 1 µM DOX (); data are those obtained at peak SMase activation (4–8 min). Results are mean ± SE of four independent experiments (three for inserts). *, P < 0.5; **, P < 0.01.

 
By using electrophoretic mobility shift assays, it has been shown that NF{kappa}B DNA-binding activity is stimulated by drugs such as DNR and mitoxantrone (13, 14, 15, 16, 17, 18, 19) , as well as by other DNA-damaging agents. However, the fact that nongenotoxic anticancer compounds, such as Vinca alkaloids and taxanes (16) , may also activate NF{kappa}B suggests that DNA damage is not required for drug-induced NF{kappa}B activation, which is essentially triggered by a stress-response pathway. In our study, we observed that both free and immobilized anthracyclines similarly activated NF{kappa}B (Fig. 5)Citation . This would mean that these agents share some NF{kappa}B-activating signaling pathways that must be generated at the cellular membrane but not in the nucleus; perhaps similarly to that observed in the UV response in which NF{kappa}B activation was clearly demonstrated to be initiated not in the nucleus but at or near the plasma membrane through a Src- and Ras-dependent mechanism (19) .



View larger version (90K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 5. NF{kappa}B-activation triggered by anthracyclines. U937 cells were either untreated (Lane 2) or stimulated for 30 min with 1 µM DNR (Lane 3), 5 µM DOX (Lane 4), 0.3 µM iDNR (Lane 5), 0.3 µM iDOX (Lane 6), 0.1 µM DNR (Lane 7), 1 µM DOX (Lane 8), or plain agarose beads (Lane 1). Nuclear fractions were prepared and equal amounts were incubated with [{gamma}32P]-ATP-labeled NF{kappa}B oligonucleotide probe. Results are representative of three independent experiments.

 
Despite many efforts, the mechanism by which anthracyclines activate NF{kappa}B is unknown. Ceramide was a plausible candidate. Indeed, previous studies have shown that the addition of ceramide induced NF{kappa}B activation (20) . As a matter of fact, we found that the inhibition of DNR-induced sphingomyelinase stimulation and subsequent ceramide production by serine-protease inhibitors reduces the capacity of the drug to activate NF{kappa}B (21) . However, because of the broad specificity of these agents, the role of ceramide was not totally established, and one could not exclude a ceramide-independent mechanism as has been proposed for TNF{alpha}-induced NF{kappa}B activation (22 , 23) . However, the present study clearly shows that anthracyclines can lead to NF{kappa}B activation in the absence of ceramide generation.

Previous studies have clearly shown that NF{kappa}B inhibits apoptosis and/or enhances survival in a large variety of hematopoietic cells, including B lymphocytes (24) , Hodgkin’s disease cells (25) , and CD34+ bone marrow cells (26) . Because of the known anti-apoptotic function of NF{kappa}B, it was tempting to speculate that this transcription factor provides significant protection against DNR-induced cytotoxicity. Such a hypothesis was supported by the fact that NF{kappa}B was found to play an essential role in preventing TNF{alpha}-induced cell death (14 , 27 , 28) . To address this question more directly for DNR, Wang et al. have used a gene construct coding for a super-repressor form of the NF{kappa}B inhibitor I{kappa}B{alpha}. These authors have shown that NF{kappa}B inhibition resulted in an increased cytotoxic effect of DNR (and ionizing radiation) in vitro (14) . Thus, one could speculate that in the absence of a ceramide-induced apoptotic signal, the remaining stress-activated NF{kappa}B survival pathway enabled the cells to present some resistance before finally giving in to drug-induced necrosis.

In conclusion, our results suggest that cellular internalization of DNR and DOX is required for apoptosis and ceramide-mediated signaling. However, we do not demonstrate unambiguously that anthracycline interaction with DNA, or more specifically with topoisomerase II, is requisite for apoptosis signaling. Nevertheless, although the immobilized drug is not the same chemical entity as the free drug, and may act by a different mechanism, one can speculate that both events (necrosis and apoptosis) are likely to be induced by the free drug (2) .

Moreover, our results may also have important clinical implications. Indeed, some pharmacological agents, including glucocorticoids, are known to inhibit cytokine-induced NF{kappa}B activation which act through induction of I{kappa}B synthesis (29 , 30) . Glucocorticoids are largely used in association with anthracyclines as part of front-line therapy of acute lymphoid leukemia and lymphoma. Whether or not glucocorticoids act in synergy with anthracyclines in lymphoid cells through a NF{kappa}B-dependent mechanism has yet to be evaluated.


   FOOTNOTES
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Supported by the Ligue Nationale Contre le Cancer and the Comités Départementaux du Gers, de l’Aveyron, et du Lot (to J-P. J.) and in part by the Faculté de Médecine Toulouse-Rangueil (to G. L.). N. M. was supported by a fellowship from the Ligue Departementale Contre le Cancer du Tarn et Garonne. Back

2 To whom requests for reprints should be addressed, at Institut National de la Santé et de la Recherche Médicale E9910, Institut Claudius Régaud, 20 rue DuPont St. Pierre, 31052 Toulouse, France. Phone: 5-61-42-41-73; Fax: 5-61-42-46-06; E-mail: jaffrezou{at}icr.fnclcc.fr Back

3 The abbreviations used are: DNR, daunorubicin; DOX, doxorubicin; iDNR, immobilized DNR; iDOX, immobilized DOX; DAPI, 4',6-diamidino-2phenylindole; PARP, poly(ADP-ribose) polymerase; AML, acute myeloid leukemia; NF{kappa}B, nuclear factor {kappa}B. Back

Received 7/24/00. Accepted 1/11/01.


   REFERENCES
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS AND DISCUSSION
 REFERENCES
 

  1. Chabner B. A., Myers C. E. Clinical pharmacology of cancer chemotherapy DeVitta V. T. J. Hellman S. Rosenberg S. A. eds. . Cancer: Principles and Practice of Oncology, : 349-395, J. B. Lippincott Co. Philadelphia 1989.
  2. Tritton T. R. Cell surface actions of Adriamycin. Pharmacol. Ther., 49: 293-309, 1991.[Medline]
  3. Potmesil M., Kirschenbaum J., Israel M., Levin M., Khetarpal V. K., Silber R. Relationship of Adriamycin concentrations to the DNA lesions induced in hypoxic and euoxic L1210 cells. Cancer Res., 43: 3528-3533, 1983.[Abstract/Free Full Text]
  4. Ross W. E., Glaubiger K., Kohn K. W. Qualitative and quantitative aspects of intercalator-induced DNA strand breaks. Biochim. Biophys. Acta, 5629: 41-50, 1979.
  5. Zwelling L. A., Kerrigan D., Michaels S. Cytotoxicity and DNA strand breaks by 5-iminodaunorubicin in mouse leukemia L1210 cells: comparison with Adriamycin and 4'-(9-acridinylamino)methanesulfon-m-aniside. Cancer Res., 42: 2687-2691, 1982.[Abstract/Free Full Text]
  6. Jaffrézou J. P., Levade T., Bettaïeb A., Andrieu N., Bezombes C., Maestre N., Vermeersch S., Rousse A., Laurent G. Daunorubicin-induced apoptosis; triggering of ceramide generation through sphingomyelin hydrolysis. EMBO J., 15: 2417-2424, 1996.[Medline]
  7. Tritton T. R., Yee G. The anticancer agent Adriamycin can be actively cytotoxic without entering cells. Science (Washington DC), 217: 248-250, 1982.[Abstract/Free Full Text]
  8. Quillet-Mary A., Mansat V., Duchayne E., Côme M. G., Allouche M., Bailly J. D., Bordier C., Laurent G. Daunorubicin-induced internucleosomal DNA fragmentation in acute myeloid cell lines. Leukemia, 10: 417-425, 1996.[Medline]
  9. Andrieu N., Salvayre R., Levade T. Evidence against involvement of the acid lysosomal sphingomyelinase in the tumor-necrosis-factor- and interleukin-1-induced sphingomyelin cycle and cell proliferation in human fibroblasts. Biochem. J., 303: 341-345, 1994.
  10. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of proteins using bicinchoninic acid. Anal. Biochem., 150: 76-85, 1985.[Medline]
  11. Folch J., Lees M., Sloane-Stanley G. H. A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem., 226: 497-509, 1957.[Free Full Text]
  12. Wiegmann K., Schütze S., Machleidt T., Witte D., Krönke M. Functional dichotomy of neutral and acidic sphingomyelinases in tumor necrosis factor signaling. Cell, 78: 1005-1008, 1994.[Medline]
  13. Piret B., Piette J. Topoisomerase poisons activate the transcription factor NF-{kappa}B in ACH-2 and CEM cells. Nucleic Acids Res., 24: 4242-4248, 1996.[Abstract/Free Full Text]
  14. Wang C. Y., Mayo M. W., Baldwin A. S., Jr. TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-{kappa} B. Science (Washington DC), 274: 784-787, 1996.[Abstract/Free Full Text]
  15. Boland M. P., Foster S. J., O’Neill L. A. J. Daunorubicin activates NF{kappa}B and induces {kappa}B-dependent gene expression in HL-60 promyelocytic and Jurkat T lymphoma cells. J. Biol. Chem., 272: 12952-12960, 1997.[Abstract/Free Full Text]
  16. Das K. C., White C. W. Activation of NF-{kappa}B by antineoplastic agents. J. Biol. Chem., 272: 14914-14920, 1997.[Abstract/Free Full Text]
  17. Hellin A. C., Calmant P., Gielin J., Bours V., Merville M. P. Nuclear factor-{kappa}B-dependent regulation of p53 gene expression induced by daunomycin genotoxic drug. Oncogene, 16: 1187-1195, 1998.[Medline]
  18. Bentires-Alj M., Hellin A. C., Ameyar M., Chouaib S., Merville M. P., Bours V. Stable inhibition of nuclear factor {kappa}B in cancer cells does not increase sensitivity to cytotoxic drugs. Cancer Res., 59: 811-815, 1999.[Abstract/Free Full Text]
  19. Devary Y., Rosette C., DiDonato J. A., Karin M. NF-{kappa}B activation by ultraviolet light not dependent on a nuclear signal. Science (Washington DC), 261: 1442-1445, 1993.[Abstract/Free Full Text]
  20. Schütze S., Potthoff K., Machleidt T., Berkovic D., Wiegmann K., Krönke M. TNF activates NF-{kappa}B by phosphatidylcholine-specific phospholipase C-induced "acidic" sphingomyelin breakdown. Cell, 71: 765-776, 1992.[Medline]
  21. Mansat V., Bettaieb A., Levade T., Laurent G., Jaffrézou J. P. Serine-protease inhibitors block neutral sphingomyelinase activation, ceramide generation, and apoptosis triggered by daunorubicin. FASEB J., 11: 695-702, 1997.[Abstract]
  22. Betts J. C., Agranoff A. B., Nabel G. J., Shayman J. A. Dissociation of endogenous cellular ceramide from NF-{kappa}B activation. J. Biol. Chem., 269: 8455-8458, 1994.[Abstract/Free Full Text]
  23. Johns L. D., Sarr T., Ranges G. E. Inhibition of ceramide pathway does not affect ability of TNF-{alpha} to activate nuclear factor-{kappa}B. J. Immunol., 152: 5877-5882, 1994.[Abstract]
  24. Wu M., Lee H., Bellas R. E., Schauer S. L., Arsura M., Katz D., Fitzgerald M. J., Rothstein T. L., Sherr D. H., Sonenshein G. E. Inhibition of NF-{kappa}B/Rel induces apoptosis of murine B cells. EMBO J., 15: 4682-4690, 1996.[Medline]
  25. Bargou R. C., Emmerich F., Krappmann D., Bomert K., Mapara M. Y., Arnold W., Royer H. D., Grinstein E., Greiner A., Scheidereit C., Dörken B. Constitutive nuclear factor-{kappa}B-RelA activation is required for proliferation and survival of Hodgkin’s disease tumor cells. J. Clin. Investig., 100: 2961-2969, 1997.[Medline]
  26. Pyatt D. W., Stillman W. S., Yang Y., Gross S., Zheng J. H., Irons R. D. An essential role for NF-{kappa}B in human CD34+ bone marrow cell survival. Blood, 93: 3302-3308, 1999.[Abstract/Free Full Text]
  27. Beg A. A., Baltimore D. An essential role for NF-{kappa}B in preventing TNF-{alpha}-induced cell death. Science (Washington DC), 274: 782-784, 1996.[Abstract/Free Full Text]
  28. Van Antwerp D. J., Martin S. J., Kafri T., Green D. R., Verma I. M. Suppression of TNF-{alpha}-induced apoptosis by NF-{kappa}B. Science (Washington DC), 274: 787-789, 1996.[Abstract/Free Full Text]
  29. Auphan N., DiDonato J. A., Rosette C., Helmberg A., Karin M. Immunosuppression by glucocorticoids: inhibition of NF-{kappa}B activity through induction of I{kappa}B synthesis. Science (Washington DC), 270: 286-290, 1995.[Abstract/Free Full Text]
  30. Scheinman R. I., Cogswell P. C., Lofquist A. K., Baldwin A. S. Role of transcriptional activation of I{kappa}B{alpha} in mediation of immunosuppression by glucocorticoids. Science (Washington DC), 270: 283-286, 1995.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 FASEB J.Home page
Y.-Y. Liu, J. Y. Yu, D. Yin, G. A. Patwardhan, V. Gupta, Y. Hirabayashi, W. M. Holleran, A. E. Giuliano, S. M. Jazwinski, V. Gouaze-Andersson, et al.
A role for ceramide in driving cancer cell resistance to doxorubicin
FASEB J, July 1, 2008; 22(7): 2541 - 2551.
[Abstract] [Full Text] [PDF]

Home page
 Mol Cancer ResHome page
C. A. Corcoran, Q. He, S. Ponnusamy, B. Ogretmen, Y. Huang, and M. S. Sheikh
Neutral Sphingomyelinase-3 Is a DNA Damage and Nongenotoxic Stress-Regulated Gene That Is Deregulated in Human Malignancies
Mol. Cancer Res., May 1, 2008; 6(5): 795 - 807.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
E. C. Attar, D. J. DeAngelo, J. G. Supko, F. D'Amato, D. Zahrieh, A. Sirulnik, M. Wadleigh, K. K. Ballen, S. McAfee, K. B. Miller, et al.
Phase I and Pharmacokinetic Study of Bortezomib in Combination with Idarubicin and Cytarabine in Patients with Acute Myelogenous Leukemia
Clin. Cancer Res., March 1, 2008; 14(5): 1446 - 1454.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
S. FOGLI, P. NIERI, and M. C. BRESCHI
The role of nitric oxide in anthracycline toxicity and prospects for pharmacologic prevention of cardiac damage
FASEB J, April 1, 2004; 18(6): 664 - 675.
[Abstract] [Full Text] [PDF]

Home page
 CROBMHome page
S. T. Sonis
THE BIOLOGIC ROLE FOR NUCLEAR FACTOR-KAPPAB IN DISEASE AND ITS POTENTIAL INVOLVEMENT IN MUCOSAL INJURY ASSOCIATED WITH ANTI-NEOPLASTIC THERAPY
Critical Reviews in Oral Biology & Medicine, September 1, 2002; 13(5): 380 - 389.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
E. Lorenzo, C. Ruiz-Ruiz, A. J. Quesada, G. Hernandez, A. Rodriguez, A. Lopez-Rivas, and J. M. Redondo
Doxorubicin Induces Apoptosis and CD95 Gene Expression in Human Primary Endothelial Cells through a p53-dependent Mechanism
J. Biol. Chem., March 22, 2002; 277(13): 10883 - 10892.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
X. Bian, L. M. McAllister-Lucas, F. Shao, K. R. Schumacher, Z. Feng, A. G. Porter, V. P. Castle, and A. W. Opipari Jr.
NF-kappa B Activation Mediates Doxorubicin-induced Cell Death in N-type Neuroblastoma Cells
J. Biol. Chem., December 21, 2001; 276(52): 48921 - 48929.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Maestre, N.
Right arrow Articles by Jaffrézou, J.-P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Maestre, N.
Right arrow Articles by Jaffrézou, J.-P.

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 Meeting Abstracts Online