[en] 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 etoposideinduced
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.
Research center :
Unité de Recherche en Biologie Cellulaire
Disciplines :
Oncology
Author, co-author :
Sermeus, Audrey
Cosse, Jean-Philippe ; Université de Liège - ULiège > GIGA-R : Epigénétique Cellulaire et Moléculaire
Crespin, Marianne
Mainfroid, Véronique
de Longueville, Françoise
Ninane, Noelle
Raes, Martine
Remacle, Jose
Michiels, Carine
Language :
English
Title :
Hypoxia induces protection against etoposide-induced apoptosis: molecular profiling of changes in gene expression and transcription factor activity
Publication date :
2008
Journal title :
Molecular Cancer
eISSN :
1476-4598
Publisher :
BioMed Central, United Kingdom
Volume :
7
Pages :
27
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FUNDP - Facultés Universitaires Notre-Dame de la Paix [BE]
Hopfl G Ogunshola O Gassmann M HIFs and tumors--causes and consequences Am J Physiol Regul Integr Comp Physiol 2004 286(4):R608-23 15003941
Semenza GL Targeting HIF-1 for cancer therapy Nat Rev Cancer 2003 3(10):721-732 10.1038/nrc1187 13130303
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 10.1096/fj.01-0944rev 12153983
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 10.1002/(SICI)1097-0215(19980821)79:4<365::AID-IJC10>3.0.CO;2-4 9699528
Harris AL Hypoxia --A key regulatory factor in tumour growth Nat Rev Cancer 2002 2(1):38-47 10.1038/nrc704 11902584
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 2334928
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 12067980
Gordan JD Simon MC Hypoxia-inducible factors: Central regulators of the tumor phenotype Curr Opin Genet Dev 2007 17(1):71-77 10.1016/ j.gde.2006.12.006 17208433
Greijer AE van der Wall E The role of hypoxia inducible factor 1 (HIF-1) in hypoxia induced apoptosis J Clin Pathol 2004 57(10):1009-1014 1770458 15452150 10.1136/jcp.2003.015032
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 10.1016/ S0006-2952(02)01155-3 12213583
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 10.1038/32925 9537326
Zhou J Schmid T Schnitzer S Brune B Tumor hypoxia and cancer progression Cancer Lett 2006 237(1):10-21 10.1016/j.canlet.2005.05.028 16002209
Bruick RK Expression of the gene encoding the proapoptotic Nip3 protein is induced by hypoxia Proc Natl Acad Sci U S A 2000 97(16):9082-9087 16825 10922063 10.1073/pnas.97.16.9082
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 387743 15082785 10.1128/MCB.24.9.3918-3927.2004
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 10.1046/j.1523-1755.2003.00301.x 14633124
Vaupel P Harrison L Tumor hypoxia: Causative factors, compensatory mechanisms, and cellular response Oncologist 2004 9(Suppl 5):4-9 10.1634/ theoncologist.9-90005-4 15591417
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 10.1074/jbc.M110629200 11864976
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):37- 388 10.1023/ A:1024177119658 12815281
Cummins EP Taylor CT Hypoxia-responsive transcription factors Pflugers Arch 2005 450(6):363-371 10.1007/s00424-005-1413-7 16007431
Appella E Anderson CW Post-translational modifications and activation of p53 by genotoxic stresses Eur J Biochem 2001 268(10):2764-2772 10.1046/ j.1432-1327.2001.02225.x 11358490
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 10.1080/15216540252774766 11795593
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 10.1074/jbc.M102075200 11352915
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 10.1074/jbc.M411858200 15611089
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 10.1006/ excr.2001.5180 11281649
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 10.1074/jbc.M306541200 14527959
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 10.1093/toxsci/kfj131 16478757
Weinmann M Belka C Plasswilm L Tumour hypoxia: Impact on biology, prognosis and treatment of solid malignant tumours Onkologie 2004 27(1):83-90 10.1159/000075611 15007254
Harrison L Blackwell K Hypoxia and anemia: Factors in decreased sensitivity to radiation therapy and chemotherapy? Oncologist 2004 9(Suppl 5):3- 40 10.1634/theoncologist.9-90005-31 15591420
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 16844113
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 10.1016/j.yexcr.2004.01.024 15093734
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
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 10.1152/ajprenal.00081.2005 15956779
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 7969076
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 9373239
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 232484 9343394
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 10.1016/S0014-5793(99)00136-2 10094470
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 10.1074/ jbc.271.51.32529 8955077
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 371100 15024076 10.1128/ MCB.24.7.2875-2889.2004
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 10.1111/j.1349-7006.2007.00537.x 17608771
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 10.1016/j.yexcr.2006.12.004 17289019
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 10.1038/sj.onc.1210008 17043658
Fels DR Koumenis C HIF-1alpha and p53: The ODD couple? Trends Biochem Sci 2005 30(8):426-429 10.1016/j.tibs.2005.06.009 15996866
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 10.1016/j.tox.2007.03.006 17416447
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 2212759 15630139 10.1084/jem.20040624
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 10807870
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 10.1089/ 107999099312821 10638705
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 10.1158/0008-5472.CAN-07-0515 17671207
Natarajan R Fisher BJ Jones DG Ghosh S Fowler AA 3rd Reoxygenating microvascular endothelium exhibits temporal dissociation of NF-kappaB and AP-1 activation Free Radic Biol Med 2002 32(10):1033-1045 10.1016/ S0891-5849(02)00813-4 12008118
Natarajan R Fisher BJ Jones DG Fowler AA 3rd 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 10.1016/S0891-5849(02)00990-5 12361806
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 10.1046/j.1471-4159.2000.0750683.x 10899943
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 2099441 17894897 10.1186/ 1476-4598-6-61
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 10.1074/ jbc.271.49.31602 8940178
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 12393189
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 10.1126/science.1589764 1589764
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 124931 12149476 10.1073/pnas.162369299
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 10.1016/ S0006-2952(02)01055-9 12106614
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 10.1093/toxsci/kfg196 12883083
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 10.1242/jcs.01651 15671065
