Cancer Research Landon Prizes for Basic and Translational Cancer Research Tumor Immunology: New Perspectives
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
[Cancer Research 59, 3053-3058, July 1, 1999]
© 1999 American Association for Cancer Research
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 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 Huang, C.
Right arrow Articles by Dong, Z.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Huang, C.
Right arrow Articles by Dong, Z.
[Cancer Research 59, 3053-3058, July 1, 1999]
© 1999 American Association for Cancer Research

Advances in Brief

Arsenic Induces Apoptosis through a c-Jun NH2-Terminal Kinase-dependent, p53-independent Pathway1

Chuanshu Huang, Wei-ya Ma, Jingxia Li and Zigang Dong2

The Hormel Institute, University of Minnesota, Austin, Minnesota 55912


   ABSTRACT
Top
 ABSTRACT
Introduction
 Materials and Methods
 REFERENCES
 
Arsenic has been used as an effective chemotherapy agent for some human cancers, such as acute promyelocytic leukemia. In this study, we found that arsenic induces activation of c-Jun NH2-terminal kinases (JNKs) at a similar dose range for induction of apoptosis in JB6 cells. In addition, we found that arsenic did not induce p53-dependent transactivation. Similarly, there was no difference in apoptosis induction between cells with p53 +/+ or p53 -/-. In contrast, arsenic-induced apoptosis was almost totally blocked by expression of a dominant-negative mutant of JNK1. These results suggest that the activation of JNKs is involved in arsenic-induced apoptosis of JB6 cells. Taken together with previous findings that p53 mutations are involved in ~50% of all human cancers and nearly all chemotherapeutic agents kill cancer cells mainly by apoptotic induction, we suggest that arsenic may be a useful agent for the treatment of cancers with p53 mutation.


   Introduction
Top
 ABSTRACT
 Introduction
Materials and Methods
 REFERENCES
 
Chronic exposure to inorganic arsenicals through contaminated drinking water, medical or other occupational exposure, is associated with increased risk of human cancers of the skin, lung, kidney, bladder, liver, and hematopoietic system (1 , 2) . Less severe dermatological effects, including hyperpigmentation and hyperkeratosis, appear on the palms and soles after arsenic exposure (3) . Epidemiological studies in humans have suggested that the cancer risk (~1%) due to arsenic in the drinking water of the United States may be comparable with that of environmental tobacco smoke and radon in homes (4) . Interestingly, arsenic-containing compounds have been used for treatment of cancer for hundreds of years in both Western and traditional Chinese medicine (5, 6, 7, 8, 9, 10) . Arsenite was routinely used to control elevated leukocyte counts in chronic myelogenous leukemia in the early 1900s (6 , 7) . Recently, arsenic trioxide (As2O3) was reported to induce complete remission in a high proportion of patients with refractory acute promyelocytic leukemia (3) . Reports from in vitro studies using NB4 cells demonstrated that arsenite-induced apoptosis correlated with down-regulation of Bcl-2 (8, 9, 10) . Therefore, although arsenic is a known carcinogen, it is also an agent that may be useful as a chemotherapeutic agent in cancer treatment.

We have reported recently that arsenic exposure in a mouse epidermal cell line induces cell transformation at only low concentration (<25 µM) but not at higher concentration (>50 µM; Ref. 11 ). Interestingly, higher concentrations of arsenic are required for activation of JNKs3 , whereas activation of Erks occurs broadly at doses ranging from 3.2 to 200 µM (11 ). More importantly, only activation of Erks and not JNKs is required to induce cell transformation, as determined by using both dominant-negative Erk2 and JNK1 (11) . These findings, taken together with the observation that arsenite exposure can induce apoptosis without differentiation in both t-RA-sensitive and t-RA-resistant promyelocytes (12, 13, 14) , raise intriguing questions about the role of activation of JNKs in cancer cell apoptosis. Therefore, this study was initiated to examine the role of JNKs and p53 in arsenic-induced apoptosis.


   Materials and Methods
Top
 ABSTRACT
 Introduction
 Materials and Methods
REFERENCES
 
Reagents.
Both arsenite and arsenate were obtained from Sigma Chemical Co.; Klenow FragEL kit was from Oncogene Research Products, Calbiochem; EGF was from Collaborative Research; luciferase assay substrate was from Promega; FBS, MEM, DMEM, RPMI 1640, and BME were from BioWhittaker; Phosphoplus MAPK antibody kit and SAPK/JNK assay kit were purchased from New England Biolabs.

Cell Culture.
JB6 P+ mouse epidermal cell line Cl 41 and its stable p53 luciferase reporter plasmid transfect, Cl 41 p53 cells, as well as dominant-negative JNK1 and its vector stable transfectants, Cl 41 DN JNK1 mass2 and Cl 41 CMV-neo massl, were cultured in monolayers at 37°C, under 5% CO2 using Eagle’s minimal essential medium containing 5% FCS, 2 mM L-glutamine, and 25 µg of gentamicin per ml (11 , 15) . Normal embryo fibroblasts (p53 +/+) or p53-deficient embryo fibroblasts (p53 -/-) were cultured in DMEM with 10% FBS, 2 mM L-glutamine, and 25 mg of gentamicin per ml (16 , 17) .

Anchorage-independent Transformation Assay.
Inhibition of arsenic on EGF-induced cell transformation was investigated in JB6 Cl 41 cells. Cells (1 x 104) were exposed to EGF (10 ng/ml) with or without different concentrations of arsenic in 0.33% BME agar containing 10% FBS over 3.5 ml of 0.5% BME agar medium containing 10% FBS. The cultures were maintained at 37°C (5% CO2 incubator) for 14–21 days. Cell colonies were scored by the methods described previously (15 , 18) .

Assay for p53-dependent Transcription Activity.
The Cl 41 p53 cell line stably expresses a luciferase gene under the control of p53 binding sequences (PG13; Ref. 16 , 19) . The cells were trypsinized, and 8 x 103 viable cells in 100 µl of 5% FBS MEM were added into each well of a 96-well plate. The cells were incubated at 37°C in a humidified atmosphere of 5% CO2. Twelve to 24 h later, cells were starved by culturing in 0.1% FBS MEM for 12 h. The cells were then exposed to UVC (60 J/m2) or different concentrations of arsenic for 24 h. The cells were extracted with lysis buffer, and luciferase activity was measured with a luminometer (Monolight 2010). Results are expressed as relative p53-dependent transcription activity (16 , 19) .

DNA Fragment End Labeling Assay.
The cells were cultured on microscope slides, treated with arsenic for 18–24 h, and then fixed. DNA fragment end labeling assays were then performed as described previously (20) with Klenow FragEL DNA fragmentation detection kit. A minimum of 200 cells was scored for the incidence of apoptosis.

JNK Activity Assay.
JNK activity assay was carried out as described previously (11 , 21) . Briefly, JB6 Cl 41 cells were starved for 48 h in 0.1% FBS MEM at 37°C in a 5% CO2 atmosphere incubator. The cells were then exposed to arsenite at concentrations and times as indicated in the figure legends. The cells were then harvested and precipitated with 2 µg of NH2-terminal c-Jun (1–89) fusion protein bound to glutathione-Sepharose beads overnight at 4°C. The beads were washed twice with 500 µl of lysis buffer with phenylmethylsulfonyl fluoride and twice with 500 µl of kinase buffer. The kinase reactions were carried out in the presence of 100 µM ATP at 30°C for 30 min. c-Jun phosphorylation was selectively measured by Western immunoblotting using a chemiluminescent detection system and specific c-Jun antibodies against phosphorylation of c-Jun at serine 63.

Phosphorylation Analysis for JNKs.
JB6 cells were seeded in six-well plates and cultured until 80–90% confluent. The cells were then starved and treated with arsenic as indicated in the JNK activity assay. After arsenic exposure, the cells were then extracted with SDS sample buffer. Immunoblot detection of phosphorylated proteins of JNKs were carried out using phospho-specific MAPK antibodies specific for phosphorylated sites on JNKs as described previously (11 , 19) .

Results
Inhibition of EGF-induced Cell Transformation by Arsenic.
Clinical studies have reported that arsenite is effective and relatively safe in the treatment of acute promyelocytic leukemia (8, 9, 10) . However, the results from both epidemiological investigations and experimental animal models have shown that arsenic is a well-documented human carcinogen (1, 2, 3, 4) . In a late-stage tumor promotion cell culture model, a mouse epidermal JB6 cell line, we reported previously that low doses of arsenite (<25 µM) could induce cell transformation by itself (11) . While studying the synergistic effect of arsenic and EGF on cell transformation efficiency, we found that relatively high doses of arsenic markedly inhibit EGF-induced cell transformation (Fig. 1)Citation . The inhibitory effects of arsenic appear to be dose dependent (Fig. 1)Citation .



View larger version (32K):
[in this window]
[in a new window]
  		Fig. 1. Inhibition of EGF- or 12-O-tetradecanoylphorobl-13-acetate-induced JB6 Cl 41 cell transformation by arsenic. JB6 Cl 41 cells (104) were exposed simultaneously to EGF (10 ng/ml) with or without different concentrations of arsenic in 0.33% BME agar containing 10% FBS over 0.5% BME agar containing 10% FBS. Cell colonies were scored after 14 days incubation at 37°C in 5% CO2. The results are expressed as described previously (15 , 18) . Each column indicates the mean of assays from triplicate experiments; bars, SD.

 
Induction of Apoptosis by Arsenic in JB6 Cells.
Evidence from both in vitro and in vivo studies have demonstrated that induction of apoptosis is involved in cancer treatment success by some chemical agents and drugs (8, 9, 10 , 12 , 14 , 16) . Because it has been reported that arsenite induces apoptosis in some leukemia cell lines (8, 9, 10 , 12 , 14) , we analyzed apoptotic induction in JB6 cells (Fig. 2)Citation . Treatment of cells with arsenite or arsenate resulted in apoptosis by 44.5 and 61.5%, respectively (Fig. 2)Citation . These results revealed that induction of apoptosis by arsenic may be associated with its inhibitory effects on EGF-induced cell transformation.



View larger version (72K):
[in this window]
[in a new window]
  		Fig. 2. Induction of apoptosis by arsenic in JB6 cells. Subconfluent (80–90%) monolayer JB6 Cl 41 cells on microscope slides were treated with medium control (a), arsenite (As3+, 200 µ M;b), or arsenate (As5+, 200 µM;c) for 15 h. Then, the cells were fixed on slides, and DNA fragment end labeling assays were performed as described in Klenow FragEL DNA fragmentation detection kit by Oncogene.

 
p53 Is Not Involved in Apoptosis Induction by Arsenic.
It is known that a number of factors and pathways can lead to apoptosis (16 , 22, 23, 24) . Many studies report that normal function of p53 is crucial for the induction of apoptosis in human and murine cells after DNA damage (16 , 22 , 25 , 26) . Apoptosis of thymocytes after irradiation was almost completely blocked in p53-deficient mice (27) . To investigate the possible role of p53 in the induction of apoptosis by arsenic, we first investigated the induction of p53-dependent transcriptional activity by arsenic in JB6 cells. We found no increase in p53-dependent transcription activity in Cl 41 p53 cells treated with arsenic at doses ranging from 12.5 to 200 µ M (Fig. 3g)Citation , whereas UVC radiation (60 J/m2) induced a >20-fold increase of p53-dependent transcription activity (Fig. 3g)Citation . These data suggested that p53 may not be involved in apoptosis induction by arsenic. To obtain direct evidence that p53 is not required for apoptosis induction by arsenic, we tested the effects of arsenic on two fibroblast cell lines, p53 +/+ and p53 -/-, derived from mouse embryos containing either wild-type p53 (p53 +/+) or were p53 deficient (p53 -/-; Ref. 16 , 17 . The results showed that treatment of cells with arsenite or arsenate for 20 h could cause apoptosis by 38.5 and 41.0% in p53 +/+ cells or by 36.5 and 34.5% in p53 -/- cells, respectively (Fig. 3, a–f). There was no difference in apoptosis induction between these two cell lines with arsenic exposure (Fig. 3, a–f)Citation . In contrast, the p53 -/- fibroblast failed to respond to phenethyl isothiocyanate and resveratrol apoptosis stimulation as described in our previous reports (16 , 20) . Thus, these results strongly argue against the involvement of p53 in arsenic-induced apoptosis.



View larger version (68K):
[in this window]
[in a new window]
  		Fig. 3. There is no requirement of p53 for apoptosis induction by arsenic. a–f, for apoptosis induction, subconfluent monolayers of p53 +/+ (a–c) or p53 -/- (d–f) cells on microscope slides were treated with 100 µM of arsenite or arsenate for 20 h. Then, the cells were fixed on slides, and DNA fragment labeling assays were performed as described in Klenow FragEL DNA fragmentation detection kit by Oncogene. a, p53 +/+ cell medium control; b, p53 +/+ cells treated with arsenite; c, p53 +/+ cells treated with arsenate; d, p53 -/- cell medium control; e, p53 -/- cells treated with arsenite; f, p53 - /- cells treated with arsenate; g, for the p53 transactivation assay, JB6, Cl 41, and PG13-luciferase (PG13-luc) stable transfectant Cl 41 p53 cells suspended in 5% FBS MEM were added to each well of 96-well plates and cultured overnight. The cells were treated with UVC (60 J/m2) or arsenic at the concentration as indicated. Twenty-four h later, the p53 activity was determined by luciferase activity assay (20) . The results are presented as relative p53 activity (16 , 19) . Bars, SD.

 
Introduction of Dominant-Negative Mutant of JNK1 Blocks the Arsenic-induced Apoptosis.
Because the activation of JNKs is suggested to play a critical role in apoptosis induction of epithelial and lymphoid cells in response to radiation, tumor necrosis factor-{alpha} or Fas ligand (23) and our findings indicate that activation of JNKs by arsenite is at the same dose range for its induction of apoptosis (11) , we hypothesized that the activation of JNKs is required for apoptosis induced by arsenic. To test this hypothesis, we first analyzed the induction of the phosphorylation of JNKs and activity by arsenic at the same induction conditions in JB6 cells. The results showed that arsenite and arsenate markedly induced both phosphorylation and activity of JNKs (Fig. 4)Citation . To further investigate the role of the activation of JNKs in arsenic-induced apoptosis, we used the well-characterized cell line, a stable transfectant of JB6 cells with dominant-negative mutant of JNK1, to test its effects on arsenic-induced apoptosis (11) . We found that arsenite-induced activation of JNKs was impaired in this cell line at all time points tested (11) . Expression of dominant-negative mutant JNK1 blocked the apoptosis induction by arsenite (4%) or arsenate (7%) as compared with vector-transfected control cells (31.5 and 40.5% for arsenite and arsenate, respectively; (Fig. 5)Citation . These data demonstrated that the pathway of JNKs plays an essential role in arsenic-induced apoptosis.



View larger version (54K):
[in this window]
[in a new window]
  		Fig. 4. Induction of JNK phosphorylation and activity by arsenic. For assay of JNK activity (A and B), Cl 41 cells were cultured in monolayers in 100-mm dishes to 90% confluency. The cells were starved by changing the medium with 0.1% FBS MEM for 48 h. The cells were then exposed to arsenic for the times and doses as indicated. The cells were harvested, and JNK activity was measured as described in "Materials and Methods." For assay of JNK phosphorylation (C), Cl 41 cells were seeded into six-well plates. After culturing at 37°C for 24 h, the cells were starved for 48 h by changing medium with 0.1% FBS MEM. Four h before cells were exposed to arsenic, the medium was changed to serum-free medium. Then, the cells were exposed to arsenite (200 µM) for the times as indicated. The cells were extracted, and JNKS, as well as phospho-JNKs proteins, were determined as described previously (11 , 19) .

 


View larger version (64K):
[in this window]
[in a new window]
  		Fig. 5. Blockade of arsenic-induced apoptosis by expression of dominant-negative mutant of JNK1. Cl 41 CMV-neo mass1 (a–c) or Cl 41 DN-JNK1 mass2 (d–f) on microscope slides were treated with 100 µM of arsenite (b and e) or arsenate (c and f) for 20 h. Then, the cells were fixed on slides, and DNA fragment labeling assays were performed as described in Klenow FragEL DNA fragmentation detection kit by Oncogene. a, Cl 41 CMV-neo mass1 medium control; d, Cl 41 DN-JNK1 mass2 medium control.

 
Discussion
The report presented here demonstrates that activation of JNKs, but not p53 protein, is the mediator of arsenic-induced apoptosis. This notion is supported by evidence from experiments using a p53-luciferase reporter stable transfectant, the p53 knockout cell line, as well as the dominant-negative mutant JNK1 stable transfectant. We found that the activation of JNKs and induction of apoptosis occur only in the cells exposed to higher doses of arsenic (>50 µM), whereas arsenic did not induce p53-dependent transcription at all doses studied. Furthermore, expression of the dominant-negative mutant of JNK1 blocks arsenic-induced cell apoptosis, whereas there is no difference for apoptosis induction by arsenic between p53 +/+ and p53 - /- cells. These data, together with our previous findings that the activation of Erks is required in arsenic-induced cell transformation, not only assess the role of MAPK members in arsenic-induced apoptosis but also provide important information in the understanding of arsenic-induced biological activity, including the effect of arsenic on cancer and in carcinogenesis. In consideration of the fact that p53 mutations are very common in human cancer and that apoptosis plays a key role in most of the chemotherapeutic agents, arsenic may potentially be very useful for the treatment of certain human cancers, especially for " drug-resistant" cancers, the effectiveness of which are through p53-dependent apoptosis induced by most of the clinically used chemotherapy agents.

Apoptosis can be initiated by a wide variety of stimuli, including developmental signals, cellular stress, disruption of cell cycle, and different kinds of chemicals (16 , 20 , 22, 23, 24 , 28) . A number of key factors involved in the regulation and coordination of apoptosis have been identified. The tumor suppressor gene, p53, has been clearly linked to the pathways leading to apoptosis in human and murine cells in response to DNA damage. This notion is supported by the findings that p53 is the most commonly mutated tumor suppressor gene, and the lack of p53 suppression or function is associated with an increased risk of tumor formation (29, 30, 31) . Transgenic mice expressing mutant p53 or p53 knockout mice with both alleles of p53 disrupted are prone to both spontaneous and induced tumors (30 , 32 , 33) . Ectopic expression of wild-type p53 in murine myeloid leukemia cells induces rapid apoptosis (34) . Thymocytes from p53 - /- mice are resistant to ionizing radiation-induced apoptosis (27) . p53 is also required for the removal of damaged keratinocytes after UV irradiation on the skin (35) . Known target genes of p53 in apoptosis are BAX, Bcl-Xl, Fas1, FASL, IGF-Bp3, PGA 608, and DR5 (22) . Because it has been suggested that arsenic induces DNA damage, such as chromosome aberration and sister chromatid exchange, we tested the possible role of p53 in arsenic-induced apoptosis. Our results indicated that p53 is not involved in apoptosis induction by arsenic because arsenic does not induce p53-depedent transcriptional activation and still initiates apoptosis in p53 - /- cells.

Recently, the pathways of JNKs were reported to be implicated in induction of apoptosis (23) . The evidence for requirement of JNKs in apoptosis derives from studies where overexpression of JNKs results in apoptosis in some cells (23) , and the antisense inhibition or the use of dominant-negative constructs attenuates the apoptotic response (23 . It was reported that in mice deficient in JNK3, which is restricted to the brain, there was a reduction in seizure activity and hippocampal apoptosis in response to the excitotoxic glutamate-receptor agonist kainic acid (36) . However, there are numerous studies providing evidence that JNK activation is either antiapoptotic or nonapoptotic (23) . The explanation for differences among different studies may be due to cell type specificity. However, consensus has not been established for either a strictly apoptotic or antiapoptotic role for the signaling of JNKs (23) . For example, it has been reported that inhibition of the signaling of JNKs does not block Fas-mediated killing in Jurkat T cells (37 , 38) , whereas it was found that Fas-mediated apoptosis in neuroblastoma cells requires the pathway of JNKs (39) . Similarly, in contrast to the evidence provided by the JNK3 knockout, data from the TRAF2 knockout mouse suggest that TRAF2 signaling of the activation of JNKs protect against tumor necrosis factor-induced apoptosis (40) . In this study, we demonstrated that arsenic-induced apoptosis is dependent on the JNKs pathway by using mouse epidermal JB6 cells stably transfected with dominant-negative mutant JNK1. Additional studies will focus on the role that ceramide generation plays in apoptosis induction by arsenic.

Nearly all chemotherapeutic drugs cause DNA damage and kill cancer cells, mainly by activating endogenous biochemical pathways for induction of tumor cell apoptosis (41 , 42) . It is known that p53-depedent apoptosis after DNA damage is mediated by the CD95 (APO-1/Fas) receptor (43) . Anticancer agents, such as cisplatin, mitomycin, methotrexate, mitoxantrone, doxorubicin, and bleomycin, at concentrations present in the sera of patients during therapy led to an up-regulation of the CD95 receptor. The up-regulation of the CD95 receptor is observed only in cells with wild-type p53 expression and not in cells with p53 mutation or that are p53 deficient (41) . Therefore, p53 inactivation is associated with resistance to anticancer chemotherapy. Because p53 is inactivated in the majority of human cancers, novel approaches to counteracting drug resistance is the goal in the development of new chemotherapy agents that are able to bypass the p53-dependent pathway and that can be specifically directed against p53-defective cancer cells (43) . In this study, we found that arsenic induces apoptosis through JNK-dependent and p53-independent pathways. Thus, arsenic is a potential treatment for cancer patients who have developed drug resistance, especially for those patients with cancers with p53 inactivation.


   ACKNOWLEDGMENTS
 
We thank Dr. Lawrence A. Donehower, Division of Molecular Virology, Baylor College of Medicine, Houston, TX, for the generous gift of p53 +/+ and p53 -/- fibroblasts; Dr. Doug Bibus for critical reading; and Andria Percival for secretarial assistance.


   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 This research was supported by The Hormel Foundation, Grant CA74916 from the National Cancer Institute, and a Grant-in-Aid from the Graduate School of the University of Minnesota. Back

2 To whom requests for reprints should be addressed, at The Hormel Institute, University of Minnesota, 801 16th Avenue NE, Austin, MN 55912. Phone: (507) 437-9640; Fax: (507) 437-9606; E-mail: zgdong{at}smig.net Back

3 The abbreviations used are: JNK, c-Jun NH2-terminal kinase; Erk, extracellular signal-regulated kinase; EGF, epidermal growth factor; P+, tumor promoter-sensitive; P-, tumor promoter-resistant; FBS, fetal bovine serum; BME, basal medium Eagle; MAPK, mitogen-activated protein kinase. Back

Received 3/ 1/99. Accepted 5/17/99.


   REFERENCES
Top
 ABSTRACT
 Introduction
 Materials and Methods
 REFERENCES
 

  1. Nriagu J. O. Arsenic in the Environment, Part II. Human Health and Ecosystem Effects, : Wiley New York 1994.
  2. Sommers S. C., McManus R. G. Multiple arsenical cancers of skin and internal organs. Cancer (Phila.), 6: 347-359, 1953.
  3. Chen C-J., Chuang Y-C., Lin T-M., Wu H-Y. Malignant neoplasms among residents of a Blackfoot disease-endemic area in Taiwan: High-arsenic artesian well water and cancers. Cancer Res., 45: 5895-5899, 1985.[Medline]
  4. Smith A. H., Hopenhayn-Rich C., Bates M. N., Goeden H. M., Hertz-Picciotto P., Duggan H. M., Wood R., Kornett M. J., Smith M. T. Cancer risks from arsenic in drinking water. Environ. Health Perspect., 97: 259-267, 1992.[Medline]
  5. Bainbridge W. S. . The Cancer Problem, : p. 271 Macmillan New York 1914.
  6. Forkner C., McNair-Scott T. F. Arsenic as a therapeutic agent in chronic myeloid leukemia. J. Am. Med. Assoc., 97: 305 1931.
  7. Kandel E. V., Leroy G. V. Chronic arsenical poisoning during the treatment of chronic myeloid leukemia. Arch. Intern. Med., 60: 846-866, 1937.
  8. Konig A., Wrazel L., Warrel R. P., Jr., Rivi R., Pandolfi P. P., Jakubowski A., Gabrilcve J. L. Comparative activity of melarsoprol and arsenic trioxide in chronic B-cell leukemia lines. Blood, 90: 562-570, 1997.[Abstract/Free Full Text]
  9. Chen G-Q., Shi X-G., Tang W., Xiong S-M., Zhu J., Cai X., Han Z-G., Ni J-H., Shi G-Y., Jia P-M., Liu M-M., He K-L., Niu C., Ma J., Zhang P., Zhang T-D., Paul P., Naoe T., Kitamura K., Miller W., Waxman S., Wang Z-Y., de The H., Chen S-J., Chen Z. Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL). I. As2O3 exerts dose-dependent dual effects on APL cells. Blood, 89: 3345-3353, 1997.[Abstract/Free Full Text]
  10. Chen G-Q., Zhu J., Shi X-G., Ni J-H., Zhong H-J., Si G-Y., Jin X-L., Tang W., Li X-S., Xong S-M., Shen Z-X., Sun G-L., Ma J., Zhang P., Zhang T-D., Gazin C., Naoe T., Chen S-J., Wang Z-Y., Chen Z. In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-Ra{alpha}/PML proteins. Blood, 88: 1052-1061, 1996.[Abstract/Free Full Text]
  11. Huang C., Ma W-Y., Li J., Goranson A., Dong Z. Requirement of Erks, but not JNKs, for arsenite-induced cell transformation. J. Biol. Chem., 274: 14595-14601, 1999.[Abstract/Free Full Text]
  12. Ludwig S. A., Hoffmeyer A., Goebeler M., Kilian K., Hafner H., Neufeld B., Han J., Rapp U. R. The stress inducer arsenite activates mitogen-activated protein kinases extracellular signal-regulated kinases 1 and 2 via a MAPK kinase 6/p38-dependent pathway. J. Biol. Chem., 273: 1917-1922, 1998.[Abstract/Free Full Text]
  13. Look A. T. Arsenic and apoptosis in the treatment of acute promyelocytic leukemia. J. Natl. Cancer Inst., 90: 86-88, 1998.[Free Full Text]
  14. Shao W., Fanelli M., Ferrara F. F., Riccioni R., Rosenauer A., Davison K., Lamph W. W., Waxman S., Pelicci P. G., Lo Coco F., Avvisati G., Testa U., Peschle C., Gambacorti-Passerini C., Nervi C., Miller W. H., Jr. Arsenic trioxide as an inducer of apoptosis and loss of PML/RAR{alpha} protein in acute promyelocytic leukemia cells. J. Natl. Cancer. Inst., 90: 124-133, 1998.[Abstract/Free Full Text]
  15. Huang C., Ma W-Y., Dong Z. Requirement of phosphatidylinositol-3 kinase in epidermal growth factor-induced AP-1 transactivation and transformation in JB6 P+ cells. Mol. Cell. Biol., 16: 6427-6435, 1996.[Abstract]
  16. Huang C., Ma W-Y., Li J., Hecht S., Dong Z. Essential role of p53 in phenethyl isothiocyanate-induced apoptosis. Cancer Res., 58: 4102-4106, 1998.[Abstract/Free Full Text]
  17. Harvey M., Sands A. T., Weiss R. S., Hegi M. E., Wiseman R. W., Pantazis P., Giovanella B. C., Tainsky M. A., Bradley A., Donehower L. A. In vitro growth characteristics of embryo fibroblasts isolated from p53-deficient mice. Oncogene, 8: 2457-2467, 1993.[Medline]
  18. Huang C., Schmid P. C., Ma W-Y., Schmid H. H. O., Dong Z. Phosphatidylinositol-3 kinase is necessary for 12-O-tetradecanoylphorbol-13-acetate-induced cell transformation and activated protein-1 activation. J. Biol. Chem., 272: 4187-4194, 1997.[Abstract/Free Full Text]
  19. Huang C., Ma W-Y., Ryan C. A., Dong Z. Proteinase inhibitors I and II from potatoes specifically block UV-induced activator protein-1 activation through a pathway that is independent of extracellular signal-regulated kinases, c-Jun N-terminal kinases, and P38 kinase. Proc. Natl. Acad. Sci. USA, 94: 11957-11962, 1997.[Abstract/Free Full Text]
  20. Huang C., Ma W-Y., Goranson A., Dong Z. Resveratrol suppresses cell transformation and induces apoptosis through a p53-dependent pathway. Carcinogenesis, 20: 237-242, 1999.[Abstract/Free Full Text]
  21. Huang C., Ma W-Y., Ding M., Bowden G. T., Dong Z. Direct evidence for an important role of sphingomyelinase in ultraviolet-induced activation of c-Jun N-terminal kinase. J. Biol. Chem., 272: 27753-27757, 1997.[Abstract/Free Full Text]
  22. Amundson S. A., Myers T. G., Fornace A. J., Jr. Roles for p53 in growth arrest and apoptosis: putting on the brakes after genotoxic stress. Oncogene, 17: 3287-3299, 1998.[Medline]
  23. Basu S., Kolesnick R. Stress signals for apoptosis: ceramide and c-Jun kinase. Oncogene, 17: 3277-3285, 1998.[Medline]
  24. Dragovich T., Rudin C. M., Thompson C. R. Signal transduction pathways that regulate cell survival and cell death. Oncogene, 17: 3207-3213, 1998.[Medline]
  25. Merritt A. J., Potten C. S., Kemp C. J., Hickman J. A., Balmain M., Lane D. P., Hall P. A. The role of p53 in spontaneous and radiation-induced apoptosis in the gastrointestinal tract of normal and p53-deficient mice. Cancer Res., 54: 614-617, 1994.[Abstract/Free Full Text]
  26. Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer drugs. Cell, 74: 957-967, 1993.[Medline]
  27. Clarke A. R., Purdie C. A., Harrison D. J., Morris R. G., Bird C. C., Hooper M. L., Wyllie A. H. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature (Lond.), 362: 849-852, 1993.[Medline]
  28. Ochi T., Nakajima F., Sakurai T., Kaise T., Oya-Ohta Y. Dimethylarsinic acid causes apoptosis in HL-60 cells via interaction with glutathione. Arch. Toxicol., 70: 815-821, 1996.[Medline]
  29. Hollstein M., Rice K., Greenbalt M. S., Soussi T., Fuchs R., Sorlie T., Hovig E., Smith-Sorenson B., Montesano R., Harris C. C. Database of p53 gene somatic mutations in human tumors and cell lines. Nucleic Acids Res., 22: 3551-3555, 1994.
  30. Donehower L. A., Harvey M., Slagle B. L., McArthur M. J., Montgomery C. A., Jr., Butel J. S., Bradley A. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumors. Nature (Lond.), 356: 215-221, 1992.[Medline]
  31. Malkin D., Li F. P., Strong L. C., Fraumeni J. F., Jr., Nelson C. E., Kim D. H., Kassel J., Gryka M. A., Bischoff F. Z., Tainsky M. A., Friend S. H. Germ line p53 mutations in a familial syndrome of breast cancer, sarcomas, and other neoplasms. Science (Washington DC), 250: 1233-1238, 1990.[Abstract/Free Full Text]
  32. Donehower L. A., Harvey M., Vogel H., McArthur M. J., Montgomery C. A. J., Park S. H., Thompson T., Ford R. J., Bradley A. Effects of genetic background on tumorigenesis in p53-deficient mice. Mol. Carcinog., 14: 16-22, 1995.[Medline]
  33. Lavigueur A., Maltby V., Mock D., Rossant J., Pawson T., Berstein A. High incidence of lung, bone, and lymphoid tumors in transgenic mice overexpressing mutant alleles of the p53 oncogene. Mol. Cell. Biol., 9: 3982-3991, 1989.[Abstract/Free Full Text]
  34. Yonish-Rouach E., Resnitzky D., Lotem J., Sachs L., Kimchi A., Oren M. Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6. Nature (Lond.), 352: 345-347, 1991.[Medline]
  35. Ziegler A., Jonason A. S., Leffell D. J., Simon J. A., Sharma H. W., Kimmelman J., Jacks T., Brash D. E. Sunburn and p53 in the onset of skin cancer. Nature (Lond.), 372: 773-776, 1994.[Medline]
  36. Yang D. D., Kuan C. Y., Whitmarsh A. J., Rincon M., Zheng T. S., Davis R. J., Rakie P., Flavell R. A. Absence of excitotoxicity-induced apoptosis in the hippocampus of mice lacking the JNK3 gene. Nature (Lond.), 389: 865-870, 1997.[Medline]
  37. Chen Y. R., Wang X. P., Templeton D., Davis R. J., Tan T. H. The role of c-Jun N-terminal kinase (JNK) in apoptosis induced by ultraviolet C and gamma radiation. Duration of JNK activation may determine cell death and proliferation. J. Biol. Chem., 271: 31929-31936, 1996.[Abstract/Free Full Text]
  38. Lenczowski J. M., Dominguez L., Eder A. M., King L. B., Zacharchuk C. M., Ashwell J. D. Lack of a role for Jun kinase and AP-1 in Fas-induced apoptosis. Mol. Cell. Biol., 17: 170-181, 1997.[Abstract]
  39. Goillot E., Raingeaud J., Ranger A., Tepper R. I., Davis R. J., Harlow E., Sanchez I. Mitogen-activated protein kinase-mediated Fas apoptotic signaling pathway. Proc. Natl. Acad. Sci. USA, 94: 3302-3307, 1997.[Abstract/Free Full Text]
  40. Yeh W. C., Shahinian A., Speiser D., Kraunus J., Billia F., Wakeham A., de la Pompa J., Ferrick D., Hum B., Iscove N., Ohashi P., Rothe M., Goeddel D., Mak T. Early lethality, functional NF-{kappa}B activation, and increased sensitivity to TNF-induced cell death in TRAF2-deficient mice. Immunity, 7: 715-725, 1997.[Medline]
  41. Miller M., Wilder S., Bannasch D., Lehlback K., Li-Weber M., Friedman S. L., Galle P. R., Stremmel W., Oren M., Krammer P. H. p53 activates the CD95 (APO-1/Fas) gene in response to DNA damage by anticancer drugs. J. Exp. Med., 188: 2033-2045, 1998.[Abstract/Free Full Text]
  42. Eldeiry W. S. The p53 pathway and cancer therapy. Cancer J., 11: 229-236, 1998.
  43. Husain A., Rosales N., Schwartz G. K., Spriggs D. R. Lisofylline sensitizes p53 mutant human ovarian carcinoma cells to the cytotoxic effects of cis-diamminedichloroplatinum (II). Gynecol. Oncol., 70: 17-22, 1998.[Medline]



This article has been cited by other articles:


Home page
 BloodHome page
E. Fox, B. I. Razzouk, B. C. Widemann, S. Xiao, M. O'Brien, W. Goodspeed, G. H. Reaman, S. M. Blaney, A. J. Murgo, F. M. Balis, et al.
Phase 1 trial and pharmacokinetic study of arsenic trioxide in children and adolescents with refractory or relapsed acute leukemia, including acute promyelocytic leukemia or lymphoma
Blood, January 15, 2008; 111(2): 566 - 573.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Biol.Home page
L. Song, J. Li, D. Zhang, Z.-g. Liu, J. Ye, Q. Zhan, H.-M. Shen, M. Whiteman, and C. Huang
IKK{beta} programs to turn on the GADD45{alpha}-MKK4-JNK apoptotic cascade specifically via p50 NF-{kappa}B in arsenite response
J. Cell Biol., November 20, 2006; 175(4): 607 - 617.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. Zhang, L. Song, J. Li, K. Wu, and C. Huang
Coordination of JNK1 and JNK2 Is Critical for GADD45{alpha} Induction and Its Mediated Cell Apoptosis in Arsenite Responses
J. Biol. Chem., November 10, 2006; 281(45): 34113 - 34123.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Ding, J. Li, C. Xue, K. Wu, W. Ouyang, D. Zhang, Y. Yan, and C. Huang
Cyclooxygenase-2 Induction by Arsenite Is through a Nuclear Factor of Activated T-cell-dependent Pathway and Plays an Antiapoptotic Role in Beas-2B Cells
J. Biol. Chem., August 25, 2006; 281(34): 24405 - 24413.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
W. Ouyang, Q. Ma, J. Li, D. Zhang, Z.-g. Liu, A. K. Rustgi, and C. Huang
Cyclin D1 Induction through I{kappa}B Kinase {beta}/Nuclear Factor-{kappa}B Pathway Is Responsible for Arsenite-Induced Increased Cell Cycle G1-S Phase Transition in Human Keratinocytes
Cancer Res., October 15, 2005; 65(20): 9287 - 9293.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
G. McCollum, P. C. Keng, J. C. States, and M. J. McCabe Jr.
Arsenite Delays Progression through Each Cell Cycle Phase and Induces Apoptosis following G2/M Arrest in U937 Myeloid Leukemia Cells
J. Pharmacol. Exp. Ther., May 1, 2005; 313(2): 877 - 887.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
L. Zhou, Y. Jing, M. Styblo, Z. Chen, and S. Waxman
Glutathione-S-transferase {pi} inhibits As2O3-induced apoptosis in lymphoma cells: involvement of hydrogen peroxide catabolism
Blood, February 1, 2005; 105(3): 1198 - 1203.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
V. N. Ivanov and T. K. Hei
Arsenite Sensitizes Human Melanomas to Apoptosis via Tumor Necrosis Factor {alpha}-mediated Pathway
J. Biol. Chem., May 21, 2004; 279(21): 22747 - 22758.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
K. Davison, K. K. Mann, S. Waxman, and W. H. Miller Jr
JNK activation is a mediator of arsenic trioxide-induced apoptosis in acute promyelocytic leukemia cells
Blood, May 1, 2004; 103(9): 3496 - 3502.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
A. T. Y. Lau, M. Li, R. Xie, Q.-Y. He, and J.-F. Chiu
Opposed arsenite-induced signaling pathways promote cell proliferation or apoptosis in cultured lung cells
Carcinogenesis, January 1, 2004; 25(1): 21 - 28.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
Y. Zhang, M. I. Dawson, Y. Ning, L. Polin, R. E. Parchment, T. Corbett, A. N. Mohamed, K.-C. Feng, L. Farhana, A. K. Rishi, et al.
Induction of apoptosis in retinoid-refractory acute myelogenous leukemia by a novel AHPN analog
Blood, November 15, 2003; 102(10): 3743 - 3752.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
D. E. Muscarella, K. A. O'Brien, A. T. Lemley, and S. E. Bloom
Reversal of Bcl-2-Mediated Resistance of the EW36 Human B-Cell Lymphoma Cell Line to Arsenite- and Pesticide-Induced Apoptosis by PK11195, a Ligand of the Mitochondrial Benzodiazepine Receptor
Toxicol. Sci., July 1, 2003; 74(1): 66 - 73.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
E. Raffoux, P. Rousselot, J. Poupon, M.-T. Daniel, B. Cassinat, R. Delarue, A.-L. Taksin, D. Rea, A. Buzyn, A. Tibi, et al.
Combined Treatment With Arsenic Trioxide and All-Trans-Retinoic Acid in Patients With Relapsed Acute Promyelocytic Leukemia
J. Clin. Oncol., June 15, 2003; 21(12): 2326 - 2334.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
E. I. Salim, H. Wanibuchi, K. Morimura, M. Wei, M. Mitsuhashi, K. Yoshida, G. Endo, and S. Fukushima
Carcinogenicity of dimethylarsinic acid in p53 heterozygous knockout and wild-type C57BL/6J mice
Carcinogenesis, February 1, 2003; 24(2): 335 - 342.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. E. Muscarella and S. E. Bloom
Cross-linking of Surface IgM in the Burkitt's Lymphoma Cell Line ST486 Provides Protection against Arsenite- and Stress-induced Apoptosis That Is Mediated by ERK and Phosphoinositide 3-Kinase Signaling Pathways
J. Biol. Chem., January 31, 2003; 278(6): 4358 - 4367.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
C. Huang, J. Li, Q. Zhang, and X. Huang
Role of Bioavailable Iron in Coal Dust-Induced Activation of Activator Protein-1 and Nuclear Factor of Activated T Cells: Difference between Pennsylvania and Utah Coal Dusts
Am. J. Respir. Cell Mol. Biol., November 1, 2002; 27(5): 568 - 574.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
C. Huang, J. Li, Q. Ke, S. S. Leonard, B.-H. Jiang, X.-S. Zhong, M. Costa, V. Castranova, and X. Shi
Ultraviolet-induced Phosphorylation of p70S6K at Thr389 and Thr421/Ser424 Involves Hydrogen Peroxide and Mammalian Target of Rapamycin but not Akt and Atypical Protein Kinase C
Cancer Res., October 15, 2002; 62(20): 5689 - 5697.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
B. J. Kim, M.-S. Kim, K.-B. Kim, K.-W. Kim, Y.-M. Hong, I.-K. Kim, H.-W. Lee, and Y.-K. Jung
Sensitizing effects of cadmium on TNF-{alpha}- and TRAIL-mediated apoptosis of NIH3T3 cells with distinct expression patterns of p53
Carcinogenesis, September 1, 2002; 23(9): 1411 - 1417.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol SciHome page
D. E. Muscarella and S. E. Bloom
Differential Activation of the c-Jun N-Terminal Kinase Pathway in Arsenite-Induced Apoptosis and Sensitization of Chemically Resistant Compared to Susceptible B-Lymphoma Cell Lines
Toxicol. Sci., July 1, 2002; 68(1): 82 - 92.
[Abstract] [Full Text] [PDF]

Home page
 The OncologistHome page
W. H. Miller Jr.
Molecular Targets of Arsenic Trioxide in Malignant Cells
Oncologist, April 1, 2002; 7(90001): 14 - 19.
[Abstract] [Full Text] [PDF]

Home page
 The OncologistHome page
M. O'Dwyer
Multifaceted Approach to the Treatment of Bcr-Abl-Positive Leukemias
Oncologist, April 1, 2002; 7(90001): 30 - 38.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Zhang, W.-Y. Ma, A. Kaji, A. M. Bode, and Z. Dong
Requirement of ATM in UVA-induced Signaling and Apoptosis
J. Biol. Chem., January 25, 2002; 277(5): 3124 - 3131.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
W. Qu, C. D. Bortner, T. Sakurai, M. J. Hobson, and M. P. Waalkes
Acquisition of apoptotic resistance in arsenic-induced malignant transformation: role of the JNK signal transduction pathway
Carcinogenesis, January 1, 2002; 23(1): 151 - 159.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
H. Maeda, S. Hori, H. Nishitoh, H. Ichijo, O. Ogawa, Y. Kakehi, and A. Kakizuka
Tumor Growth Inhibition by Arsenic Trioxide (As2O3) in the Orthotopic Metastasis Model of Androgen-independent Prostate Cancer
Cancer Res., July 1, 2001; 61(14): 5432 - 5440.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
N. Chen, M. Nomura, Q.-B. She, W.-Y. Ma, A. M. Bode, L. Wang, R. A. Flavell, and Z. Dong
Suppression of Skin Tumorigenesis in c-Jun NH2-Terminal Kinase-2-Deficient Mice
Cancer Res., May 1, 2001; 61(10): 3908 - 3912.
[Abstract] [Full Text]

Home page
 J. Pharmacol. Exp. Ther.Home page
M. J. McCabe Jr., K. P. Singh, S. A. Reddy, B. Chelladurai, J. G. Pounds, J. J. Reiners Jr., and J. C. States
Sensitivity of Myelomonocytic Leukemia Cells to Arsenite-Induced Cell Cycle Disruption, Apoptosis, and Enhanced Differentiation Is Dependent on the Inter-Relationship between Arsenic Concentration, Duration of Treatment, and Cell Cycle Phase
J. Pharmacol. Exp. Ther., November 1, 2000; 295(2): 724 - 733.
[Abstract] [Full Text]

Home page
 Cancer Res.Home page
S. Mandlekar, R. Yu, T.-H. Tan, and A.-N. T. Kong
Activation of Caspase-3 and c-Jun NH2-terminal Kinase-1 Signaling Pathways in Tamoxifen-induced Apoptosis of Human Breast Cancer Cells
Cancer Res., November 1, 2000; 60(21): 5995 - 6000.
[Abstract] [Full Text]

Home page
 J. Immunol.Home page
K. Hossain, A. A. Akhand, M. Kato, J. Du, K. Takeda, J. Wu, K. Takeuchi, W. Liu, H. Suzuki, and I. Na