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Molecular Cancer
Published in: Molecular Cancer 1/2008

Open Access 01-12-2008 | Research

Hypoxia induces protection against etoposide-induced apoptosis: molecular profiling of changes in gene expression and transcription factor activity

Authors: Audrey Sermeus, Jean-Philippe Cosse, Marianne Crespin, Veronique Mainfroid, Francoise de Longueville, Noelle Ninane, Martine Raes, Jose Remacle, Carine Michiels

Published in: Molecular Cancer | Issue 1/2008

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Abstract

Background

it is now well established that hypoxia renders tumor cells resistant to radio- but also chemotherapy. However, few elements are currently available as for the mechanisms underlying this protection.

Results

in this study, physiological hypoxia was shown to inhibit apoptosis induced in HepG2 cells by etoposide. Indeed, hypoxia reduced DNA fragmentation, caspase activation and PARP cleavage. The DNA binding activity of 10 transcription factors was followed while the actual transcriptional activity was measured using specific reporter plasmids. Of note is the inhibition of the etoposide-induced activation of p53 under hypoxia. In parallel, data from low density DNA microarrays indicate that the expression of several pro- and anti-apoptotic genes was modified, among which are Bax and Bak whose expression profile paralleled p53 activity. Cluster analysis of data unravels several possible pathways involved in the hypoxia-induced protection against etoposide-induced apoptosis: one of them could be the inhibition of p53 activity under hypoxia since caspase 3 activity parallels Bax and Bak expression profile. Moreover, specific downregulation of HIF-1α by RNA interference significantly enhanced apoptosis under hypoxia possibly by preventing the hypoxia mediated decrease in Bak expression without altering Bax expression.

Conclusion

these results are a clear demonstration that hypoxia has a direct protective effect on apoptotic cell death. Moreover, molecular profiling points to putative pathways responsible for tumor growth in challenging environmental conditions and cancer cell resistance to chemotherapeutic agents.
Appendix
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Literature
1.
Hopfl G, Ogunshola O, Gassmann M: HIFs and tumors--causes and consequences. Am J Physiol Regul Integr Comp Physiol. 2004, 286 (4): R608-23.CrossRefPubMed
2.
3.
Wenger RH: Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. Faseb J. 2002, 16 (10): 1151-1162.CrossRefPubMed
4.
Hockel M, Schlenger K, Hockel S, Aral B, Schaffer U, Vaupel P: Tumor hypoxia in pelvic recurrences of cervical cancer. Int J Cancer. 1998, 79 (4): 365-369.CrossRefPubMed
5.
6.
Teicher BA, Holden SA, al-Achi A, Herman TS: Classification of antineoplastic treatments by their differential toxicity toward putative oxygenated and hypoxic tumor subpopulations in vivo in the FSaIIC murine fibrosarcoma. Cancer Res. 1990, 50 (11): 3339-3344.PubMed
7.
Comerford KM, Wallace TJ, Karhausen J, Louis NA, Montalto MC, Colgan SP: Hypoxia-inducible factor-1-dependent regulation of the multidrug resistance (MDR1) gene. Cancer Res. 2002, 62 (12): 3387-3394.PubMed
8.
9.
10.
Piret JP, Mottet D, Raes M, Michiels C: Is HIF-1alpha a pro- or an anti-apoptotic protein?. Biochem Pharmacol. 2002, 64 (5-6): 889-892.CrossRefPubMed
11.
An WG, Kanekal M, Simon MC, Maltepe E, Blagosklonny MV, Neckers LM: Stabilization of wild-type p53 by hypoxia-inducible factor 1alpha. Nature. 1998, 392 (6674): 405-408.CrossRefPubMed
12.
Zhou J, Schmid T, Schnitzer S, Brune B: Tumor hypoxia and cancer progression. Cancer Lett. 2006, 237 (1): 10-21.CrossRefPubMed
13.
14.
Lee MJ, Kim JY, Suk K, Park JH: Identification of the hypoxia-inducible factor 1 alpha-responsive HGTD-P gene as a mediator in the mitochondrial apoptotic pathway. Mol Cell Biol. 2004, 24 (9): 3918-3927.PubMedCentralCrossRefPubMed
15.
Tanaka T, Miyata T, Inagi R, Kurokawa K, Adler S, Fujita T, Nangaku M: Hypoxia-induced apoptosis in cultured glomerular endothelial cells: involvement of mitochondrial pathways. Kidney Int. 2003, 64 (6): 2020-2032.CrossRefPubMed
16.
Vaupel P, Harrison L: Tumor hypoxia: causative factors, compensatory mechanisms, and cellular response. Oncologist. 2004, 9 Suppl 5: 4-9.CrossRefPubMed
17.
Karpinich NO, Tafani M, Rothman RJ, Russo MA, Farber JL: The course of etoposide-induced apoptosis from damage to DNA and p53 activation to mitochondrial release of cytochrome c. J Biol Chem. 2002, 277 (19): 16547-16552.CrossRefPubMed
18.
Bjorling-Poulsen M, Issinger OG: cDNA array analysis of alterations in gene expression in the promyelocytic leukemia cell line, HL-60, after apoptosis induction with etoposide. Apoptosis. 2003, 8 (4): 377-388.CrossRefPubMed
19.
Cummins EP, Taylor CT: Hypoxia-responsive transcription factors. Pflugers Arch. 2005, 450 (6): 363-371.CrossRefPubMed
20.
Appella E, Anderson CW: Post-translational modifications and activation of p53 by genotoxic stresses. Eur J Biochem. 2001, 268 (10): 2764-2772.CrossRefPubMed
21.
Michiels C, Minet E, Michel G, Mottet D, Piret JP, Raes M: HIF-1 and AP-1 cooperate to increase gene expression in hypoxia: role of MAP kinases. IUBMB Life. 2001, 52 (1-2): 49-53.CrossRefPubMed
22.
Potapova O, Basu S, Mercola D, Holbrook NJ: Protective role for c-Jun in the cellular response to DNA damage. J Biol Chem. 2001, 276 (30): 28546-28553.CrossRefPubMed
23.
Piret JP, Minet E, Cosse JP, Ninane N, Debacq C, Raes M, Michiels C: Hypoxia-inducible factor-1-dependent overexpression of myeloid cell factor-1 protects hypoxic cells against tert-butyl hydroperoxide-induced apoptosis. J Biol Chem. 2005, 280 (10): 9336-9344.CrossRefPubMed
24.
Minet E, Michel G, Mottet D, Piret JP, Barbieux A, Raes M, Michiels C: c-JUN gene induction and AP-1 activity is regulated by a JNK-dependent pathway in hypoxic HepG2 cells. Exp Cell Res. 2001, 265 (1): 114-124.CrossRefPubMed
25.
Wang Q, Wang X, Evers BM: Induction of cIAP-2 in human colon cancer cells through PKC delta/NF-kappa B. J Biol Chem. 2003, 278 (51): 51091-51099.CrossRefPubMed
26.
Xie J, Shaikh ZA: Cadmium-induced apoptosis in rat kidney epithelial cells involves decrease in nuclear factor-kappa B activity. Toxicol Sci. 2006, 91 (1): 299-308.CrossRefPubMed
27.
Weinmann M, Belka C, Plasswilm L: Tumour hypoxia: impact on biology, prognosis and treatment of solid malignant tumours. Onkologie. 2004, 27 (1): 83-90.CrossRefPubMed
28.
Harrison L, Blackwell K: Hypoxia and anemia: factors in decreased sensitivity to radiation therapy and chemotherapy?. Oncologist. 2004, 9 Suppl 5: 31-40.CrossRefPubMed
29.
Piret JP, Cosse JP, Ninane N, Raes M, Michiels C: Hypoxia protects HepG2 cells against etoposide-induced apoptosis VIA a HIF-1-independent pathway. Exp Cell Res. 2006
30.
Piret JP, Lecocq C, Toffoli S, Ninane N, Raes M, Michiels C: Hypoxia and CoCl2 protect HepG2 cells against serum deprivation- and t-BHP-induced apoptosis: a possible anti-apoptotic role for HIF-1. Exp Cell Res. 2004, 295 (2): 340-349.CrossRefPubMed
31.
Song X, Liu X, Chi W, Liu Y, Wei L, Wang X, Yu J: Hypoxia-induced resistance to cisplatin and doxorubicin in non-small cell lung cancer is inhibited by silencing of HIF-1alpha gene. Cancer Chemother Pharmacol. 2006
32.
Tanaka T, Kojima I, Ohse T, Inagi R, Miyata T, Ingelfinger JR, Fujita T, Nangaku M: Hypoxia-inducible factor modulates tubular cell survival in cisplatin nephrotoxicity. Am J Physiol Renal Physiol. 2005, 289 (5): F1123-33.CrossRefPubMed
33.
Mizumoto K, Rothman RJ, Farber JL: Programmed cell death (apoptosis) of mouse fibroblasts is induced by the topoisomerase II inhibitor etoposide. Mol Pharmacol. 1994, 46 (5): 890-895.PubMed
34.
Martins LM, Mesner PW, Kottke TJ, Basi GS, Sinha S, Tung JS, Svingen PA, Madden BJ, Takahashi A, McCormick DJ, Earnshaw WC, Kaufmann SH: Comparison of caspase activation and subcellular localization in HL-60 and K562 cells undergoing etoposide-induced apoptosis. Blood. 1997, 90 (11): 4283-4296.PubMed
35.
Bian J, Sun Y: Transcriptional activation by p53 of the human type IV collagenase (gelatinase A or matrix metalloproteinase 2) promoter. Mol Cell Biol. 1997, 17 (11): 6330-6338.PubMedCentralPubMed
36.
Wang Y, Rea T, Bian J, Gray S, Sun Y: Identification of the genes responsive to etoposide-induced apoptosis: application of DNA chip technology. FEBS Lett. 1999, 445 (2-3): 269-273.CrossRefPubMed
37.
Semenza GL, Jiang BH, Leung SW, Passantino R, Concordet JP, Maire P, Giallongo A: Hypoxia response elements in the aldolase A, enolase 1, and lactate dehydrogenase A gene promoters contain essential binding sites for hypoxia-inducible factor 1. J Biol Chem. 1996, 271 (51): 32529-32537.CrossRefPubMed
38.
Erler JT, Cawthorne CJ, Williams KJ, Koritzinsky M, Wouters BG, Wilson C, Miller C, Demonacos C, Stratford IJ, Dive C: Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance. Mol Cell Biol. 2004, 24 (7): 2875-2889.PubMedCentralCrossRefPubMed
39.
Zeng L, Kizaka-Kondoh S, Itasaka S, Xie X, Inoue M, Tanimoto K, Shibuya K, Hiraoka M: Hypoxia inducible factor-1 influences sensitivity to paclitaxel of human lung cancer cell lines under normoxic conditions. Cancer Sci. 2007, 98 (9): 1394-1401.CrossRefPubMed
40.
Fang Y, Sullivan R, Graham CH: Confluence-dependent resistance to doxorubicin in human MDA-MB-231 breast carcinoma cells requires hypoxia-inducible factor-1 activity. Exp Cell Res. 2007, 313 (5): 867-877.CrossRefPubMed
41.
Kilic M, Kasperczyk H, Fulda S, Debatin KM: Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance. Oncogene. 2007, 26 (14): 2027-2038.CrossRefPubMed
42.
Fels DR, Koumenis C: HIF-1alpha and p53: the ODD couple?. Trends Biochem Sci. 2005, 30 (8): 426-429.CrossRefPubMed
43.
Fong CC, Zhang Q, Shi YF, Wu RS, Fong WF, Yang M: Effect of hypoxia on RAW264.7 macrophages apoptosis and signaling. Toxicology. 2007, 235 (1-2): 52-61.CrossRefPubMed
44.
Walmsley SR, Print C, Farahi N, Peyssonnaux C, Johnson RS, Cramer T, Sobolewski A, Condliffe AM, Cowburn AS, Johnson N, Chilvers ER: Hypoxia-induced neutrophil survival is mediated by HIF-1alpha-dependent NF-kappaB activity. J Exp Med. 2005, 201 (1): 105-115.PubMedCentralCrossRefPubMed
45.
Matsushita H, Morishita R, Nata T, Aoki M, Nakagami H, Taniyama Y, Yamamoto K, Higaki J, Yasufumi K, Ogihara T: Hypoxia-induced endothelial apoptosis through nuclear factor-kappaB (NF-kappaB)-mediated bcl-2 suppression: in vivo evidence of the importance of NF-kappaB in endothelial cell regulation. Circ Res. 2000, 86 (9): 974-981.CrossRefPubMed
46.
Shi Q, Le X, Abbruzzese JL, Wang B, Mujaida N, Matsushima K, Huang S, Xiong Q, Xie K: Cooperation between transcription factor AP-1 and NF-kappaB in the induction of interleukin-8 in human pancreatic adenocarcinoma cells by hypoxia. J Interferon Cytokine Res. 1999, 19 (12): 1363-1371.CrossRefPubMed
47.
Lluis JM, Buricchi F, Chiarugi P, Morales A, Fernandez-Checa JC: Dual role of mitochondrial reactive oxygen species in hypoxia signaling: activation of nuclear factor-{kappa}B via c-SRC and oxidant-dependent cell death. Cancer Res. 2007, 67 (15): 7368-7377.CrossRefPubMed
48.
Natarajan R, Fisher BJ, Jones DG, Ghosh S, Fowler AA: Reoxygenating microvascular endothelium exhibits temporal dissociation of NF-kappaB and AP-1 activation. Free Radic Biol Med. 2002, 32 (10): 1033-1045.CrossRefPubMed
49.
Natarajan R, Fisher BJ, Jones DG, Fowler AA: Atypical mechanism of NF-kappaB activation during reoxygenation stress in microvascular endothelium: a role for tyrosine kinases. Free Radic Biol Med. 2002, 33 (7): 962-CrossRefPubMed
50.
Tamatani M, Mitsuda N, Matsuzaki H, Okado H, Miyake S, Vitek MP, Yamaguchi A, Tohyama M: A pathway of neuronal apoptosis induced by hypoxia/reoxygenation: roles of nuclear factor-kappaB and Bcl-2. J Neurochem. 2000, 75 (2): 683-693.CrossRefPubMed
51.
Cosse JP, Sermeus A, Vannuvel K, Ninane N, Raes M, Michiels C: Differential effects of hypoxia on etoposide-induced apoptosis according to the cancer cell lines. Mol Cancer. 2007, 6 (1): 61-PubMedCentralCrossRefPubMed
52.
Chen Y, Takeshita A, Ozaki K, Kitano S, Hanazawa S: Transcriptional regulation by transforming growth factor beta of the expression of retinoic acid and retinoid X receptor genes in osteoblastic cells is mediated through AP-1. J Biol Chem. 1996, 271 (49): 31602-31606.CrossRefPubMed
53.
Michel G, Minet E, Mottet D, Remacle J, Michiels C: Site-directed mutagenesis studies of the hypoxia-inducible factor-1alpha DNA-binding domain. Biochim Biophys Acta. 2002, 1578 (1-3): 73-83.CrossRefPubMed
54.
Kern SE, Pietenpol JA, Thiagalingam S, Seymour A, Kinzler KW, Vogelstein B: Oncogenic forms of p53 inhibit p53-regulated gene expression. Science. 1992, 256 (5058): 827-830.CrossRefPubMed
55.
James L, Eisenman RN: Myc and Mad bHLHZ domains possess identical DNA-binding specificities but only partially overlapping functions in vivo. Proc Natl Acad Sci U S A. 2002, 99 (16): 10429-10434.PubMedCentralCrossRefPubMed
56.
de Longueville F, Surry D, Meneses-Lorente G, Bertholet V, Talbot V, Evrard S, Chandelier N, Pike A, Worboys P, Rasson JP, Le Bourdelles B, Remacle J: Gene expression profiling of drug metabolism and toxicology markers using a low-density DNA microarray. Biochem Pharmacol. 2002, 64 (1): 137-149.CrossRefPubMed
57.
de Longueville F, Atienzar FA, Marcq L, Dufrane S, Evrard S, Wouters L, Leroux F, Bertholet V, Gerin B, Whomsley R, Arnould T, Remacle J, Canning M: Use of a low-density microarray for studying gene expression patterns induced by hepatotoxicants on primary cultures of rat hepatocytes. Toxicol Sci. 2003, 75 (2): 378-392.CrossRefPubMed
58.
Debacq-Chainiaux F, Borlon C, Pascal T, Royer V, Eliaers F, Ninane N, Carrard G, Friguet B, de Longueville F, Boffe S, Remacle J, Toussaint O: Repeated exposure of human skin fibroblasts to UVB at subcytotoxic level triggers premature senescence through the TGF-beta1 signaling pathway. J Cell Sci. 2005, 118 (Pt 4): 743-758.CrossRefPubMed
59.
Metadata
Title
Hypoxia induces protection against etoposide-induced apoptosis: molecular profiling of changes in gene expression and transcription factor activity
Authors
Audrey Sermeus
Jean-Philippe Cosse
Marianne Crespin
Veronique Mainfroid
Francoise de Longueville
Noelle Ninane
Martine Raes
Jose Remacle
Carine Michiels
Publication date
01-12-2008
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2008
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-7-27

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