BNIP3 protects HepG2 cells against etoposide-induced cell death under hypoxia by an autophagy-independent pathway

[en] Tumor hypoxia is a common characteristic of most solid tumors and is correlated with poor prognosis for patients partly because hypoxia promotes resistance to cancer therapy. Hypoxia selects cancer cells that are resistant to apoptosis and allows the onset of mechanisms that promote cancer cells survival including autophagy. Previously, we showed that human hepatoma HepG2 cells were protected under hypoxia against the etoposide-induced apoptosis. In this study, respective putative contribution of autophagy and BNIP3 in the protection conferred by hypoxia against the etoposide-induced apoptosis was investigated. We report that autophagy is induced by etoposide, a process that is not affected by hypoxic conditions. Using Atg5 siRNA, we show that etoposide-induced autophagy promotes apoptotic cell death under normoxia but not under hypoxia. Then, we investigated whether the hypoxia-induced protein BNIP3 could explain the different effect of autophagy on cell death under hypoxia or normoxia. We show that the silencing of BNIP3 does not affect autophagy whatever the pO2 but participates in the protective effect of hypoxia against etoposide-induced apoptosis. Together, these results suggest that autophagy might be involved in etoposide-induced cell death only under normoxia and that BNIP3 is a major effector of the protective mechanism conferred by hypoxia to protect cancer cells against etoposide-induced apoptotic cell death.

Research Center/Unit :

Unité de Recherche en Biologie Cellulaire

Disciplines :

Oncology

Author, co-author :

Cosse, Jean-Philippe ; Université de Liège - ULiège > GIGA-R : Epigénétique Cellulaire et Moléculaire

Rommelaere, Guillaume

Ninane, Noelle

Arnould, Thierry

Michiels, Carine

Language :

English

Title :

BNIP3 protects HepG2 cells against etoposide-induced cell death under hypoxia by an autophagy-independent pathway

Publication date :

2010

Journal title :

Biochemical Pharmacology

ISSN :

0006-2952

eISSN :

1873-2968

Publisher :

Elsevier Science, Oxford, United Kingdom

Volume :

Pages :

1160-1169

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

FUNDP - Facultés Universitaires Notre-Dame de la Paix

Available on ORBi :

since 15 February 2011

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Bibliography

Rockwell S. Oxygen delivery: implications for the biology and therapy of solid tumors. Oncol Res 1997, 9:383-390.
Vaupel P., Mayer A. Hypoxia and anemia: effects on tumor biology and treatment resistance. Transfus Clin Biol 2005, 12:5-10.
Tredan O., Galmarini C.M., Patel K., Tannock I.F. Drug resistance and the solid tumor microenvironment. J Natl Cancer Inst 2007, 99:1441-1454.
Cosse J.P., Michiels C. Tumour hypoxia affects the responsiveness of cancer cells to chemotherapy and promotes cancer progression. Anticancer Agents Med Chem 2008, 8:790-797.
Erler J.T., Cawthorne C.J., Williams K.J., Koritzinsky M., Wouters B.G., Wilson C., et al. 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, 24:2875-2889.
Kim J.Y., Ahn H.J., Ryu J.H., Suk K., Park J.H. BH3-only protein Noxa is a mediator of hypoxic cell death induced by hypoxia-inducible factor 1alpha. J Exp Med 2004, 199:113-124.
Peng X.H., Karna P., Cao Z., Jiang B.H., Zhou M., Yang L. Cross-talk between epidermal growth factor receptor and hypoxia-inducible factor-1alpha signal pathways increases resistance to apoptosis by up-regulating survivin gene expression. J Biol Chem 2006, 281:25903-25914.
Piret J.P., Minet E., Cosse J.P., Ninane N., Debacq C., Raes M., et al. Hypoxia-inducible factor-1-dependent overexpression of myeloid cell factor-1 protects hypoxic cells against tert-butyl hydroperoxide-induced apoptosis. J Biol Chem 2005, 280:9336-9344.
Graeber T.G., Osmanian C., Jacks T., Housman D.E., Koch C.J., Lowe S.W., et al. Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature 1996, 379:88-91.
Zhang H., Bosch-Marce M., Shimoda L.A., Tan Y.S., Baek J.H., Wesley J.B., et al. Mitochondrial autophagy is an HIF-1-dependent adaptive metabolic response to hypoxia. J Biol Chem 2008, 283:10892-10903.
Scarlatti F., Granata R., Meijer A.J., Codogno P. Does autophagy have a license to kill mammalian cells?. Cell Death Differ 2009, 16:12-20.
Chen G., Cizeau J., Vande Velde C., Park J.H., Bozek G., Bolton J., et al. Nix and Nip3 form a subfamily of pro-apoptotic mitochondrial proteins. J Biol Chem 1999, 274:7-10.
Regula K.M., Ens K., Kirshenbaum L.A. Inducible expression of BNIP3 provokes mitochondrial defects and hypoxia-mediated cell death of ventricular myocytes. Circ Res 2002, 91:226-231.
Kubli D.A., Ycaza J.E., Gustafsson A.B. Bnip3 mediates mitochondrial dysfunction and cell death through Bax and Bak. Biochem J 2007, 405:407-415.
Zhang Z., Yang X., Zhang S., Ma X., Kong J. BNIP3 upregulation and EndoG translocation in delayed neuronal death in stroke and in hypoxia. Stroke 2007, 38:1606-1613.
Azad M.B., Chen Y., Henson E.S., Cizeau J., McMillan-Ward E., Israels S.J., et al. Hypoxia induces autophagic cell death in apoptosis-competent cells through a mechanism involving BNIP3. Autophagy 2008, 4:195-204.
Li Y., Wang Y., Kim E., Beemiller P., Wang C.Y., Swanson J., et al. Bnip3 mediates the hypoxia-induced inhibition on mammalian target of rapamycin by interacting with Rheb. J Biol Chem 2007, 282:35803-35813.
Tracy K., Macleod K.F. Regulation of mitochondrial integrity, autophagy and cell survival by BNIP3. Autophagy 2007, 3:616-619.
Lozano J., Menendez S., Morales A., Ehleiter D., Liao W.C., Wagman R., et al. Cell autonomous apoptosis defects in acid sphingomyelinase knockout fibroblasts. J Biol Chem 2001, 276:442-448.
Wellington C.L., Ellerby L.M., Hackam A.S., Margolis R.L., Trifiro M.A., Singaraja R., et al. Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J Biol Chem 1998, 273:9158-9167.
Cosse J.P., 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, 6:61.
Cosse J.P., Ronvaux M., Ninane N., Raes M.J., Michiels C. Hypoxia-induced decrease in p53 protein level and increase in c-jun DNA binding activity results in cancer cell resistance to etoposide. Neoplasia 2009, 11:976-986.
Sermeus A., Cosse J.P., Crespin M., Mainfroid V., de Longueville F., Ninane N., et al. Hypoxia induces protection against etoposide-induced apoptosis: molecular profiling of changes in gene expression and transcription factor activity. Mol Cancer 2008, 7:27.
Takeuchi H., Kondo Y., Fujiwara K., Kanzawa T., Aoki H., Mills G.B., et al. Synergistic augmentation of rapamycin-induced autophagy in malignant glioma cells by phosphatidylinositol 3-kinase/protein kinase B inhibitors. Cancer Res 2005, 65:3336-3346.
Kanzawa T., Germano I.M., Komata T., Ito H., Kondo Y., Kondo S. Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ 2004, 11:448-457.
Biederbick A., Kern H.F., Elsasser H.P. Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles. Eur J Cell Biol 1995, 66:3-14.
Munafo D.B., Colombo M.I. A novel assay to study autophagy: regulation of autophagosome vacuole size by amino acid deprivation. J Cell Sci 2001, 114:3619-3629.
Bampton E.T., Goemans C.G., Niranjan D., Mizushima N., Tolkovsky A.M. The dynamics of autophagy visualized in live cells: from autophagosome formation to fusion with endo/lysosomes. Autophagy 2005, 1:23-36.
Klionsky D.J., Abeliovich H., Agostinis P., Agrawal D.K., Aliev G., Askew D.S., et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 2008, 4:151-175.
Ichimura Y., Imamura Y., Emoto K., Umeda M., Noda T., Ohsumi Y. In vivo and in vitro reconstitution of Atg8 conjugation essential for autophagy. J Biol Chem 2004, 279:40584-40592.
Mizushima N., Yoshimori T. How to interpret LC3 immunoblotting. Autophagy 2007, 3:542-545.
Tanida I., Minematsu-Ikeguchi N., Ueno T., Kominami E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 2005, 1:84-91.
Sowter H.M., Ratcliffe P.J., Watson P., Greenberg A.H., Harris A.L. HIF-1-dependent regulation of hypoxic induction of the cell death factors BNIP3 and NIX in human tumors. Cancer Res 2001, 61:6669-6673.
Yang Y.P., Liang Z.Q., Gu Z.L., Qin Z.H. Molecular mechanism and regulation of autophagy. Acta Pharmacol Sin 2005, 26:1421-1434.
Elmore S.P., Qian T., Grissom S.F., Lemasters J.J. The mitochondrial permeability transition initiates autophagy in rat hepatocytes. FASEB J 2001, 15:2286-2287.
Clarke P.G. Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl) 1990, 181:195-213.
Bursch W. The autophagosomal-lysosomal compartment in programmed cell death. Cell Death Differ 2001, 8:569-581.
Lemasters J.J., Nieminen A.L., Qian T., Trost L.C., Elmore S.P., Nishimura Y., et al. The mitochondrial permeability transition in cell death: a common mechanism in necrosis, apoptosis and autophagy. Biochim Biophys Acta 1998, 1366:177-196.
Boya P., Gonzalez-Polo R.A., Casares N., Perfettini J.L., Dessen P., Larochette N., et al. Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol 2005, 25:1025-1040.
Ravikumar B., Berger Z., Vacher C., O'Kane C.J., Rubinsztein D.C. Rapamycin pre-treatment protects against apoptosis. Hum Mol Genet 2006, 15:1209-1216.
Espert L., Denizot M., Grimaldi M., Robert-Hebmann V., Gay B., Varbanov M., et al. Autophagy is involved in T cell death after binding of HIV-1 envelope proteins to CXCR4. J Clin Invest 2006, 116:2161-2172.
Abedin M.J., Wang D., McDonnell M.A., Lehmann U., Kelekar A. Autophagy delays apoptotic death in breast cancer cells following DNA damage. Cell Death Differ 2007, 14:500-510.
Hoyer-Hansen M., Bastholm L., Mathiasen I.S., Elling F., Jaattela M. Vitamin D analog EB1089 triggers dramatic lysosomal changes and Beclin 1-mediated autophagic cell death. Cell Death Differ 2005, 12:1297-1309.
Kanzawa T., Zhang L., Xiao L., Germano I.M., Kondo Y., Kondo S. Arsenic trioxide induces autophagic cell death in malignant glioma cells by upregulation of mitochondrial cell death protein BNIP3. Oncogene 2005, 24:980-991.
Kaushal G.P., Kaushal V., Herzog C., Yang C. Autophagy delays apoptosis in renal tubular epithelial cells in cisplatin cytotoxicity. Autophagy 2008, 4:710-712.
Qadir M.A., Kwok B., Dragowska W.H., To K.H., Le D., Bally M.B., et al. Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization. Breast Cancer Res Treat 2008, 112:389-403.
Shimizu S., Kanaseki T., Mizushima N., Mizuta T., Arakawa-Kobayashi S., Thompson C.B., et al. Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes. Nat Cell Biol 2004, 6:1221-1228.
Munoz-Gamez J.A., Rodriguez-Vargas J.M., Quiles-Perez R., Aguilar-Quesada R., Martin-Oliva D., de Murcia G., et al. PARP-1 is involved in autophagy induced by DNA damage. Autophagy 2009, 5:61-74.
Papandreou I., Lim A.L., Laderoute K., Denko N.C. Hypoxia signals autophagy in tumor cells via AMPK activity, independent of HIF-1, BNIP3, and BNIP3L. Cell Death Differ 2008, 15:1572-1581.
Rouschop K.M, Wouters B.G. Regulation of autophagy through multiple independent hypoxic signaling pathways. Curr Mol Med 2009, 9:417-424.
Longo L., Platini F., Scardino A., Alabiso O., Vasapollo G., Tessitore L. Autophagy inhibition enhances anthocyanin-induced apoptosis in hepatocellular carcinoma. Mol Cancer Ther 2008, 7:2476-2485.
Yokoyama T., Miyazawa K., Naito M., Toyotake J., Tauchi T., Itoh M., et al. Vitamin K2 induces autophagy and apoptosis simultaneously in leukemia cells. Autophagy 2008, 4:629-640.
Yousefi S., Perozzo R., Schmid I., Ziemiecki A., Schaffner T., Scapozza L., et al. Calpain-mediated cleavage of Atg5 switches autophagy to apoptosis. Nat Cell Biol 2006, 8:1124-1132.
Vande Velde C., Cizeau J., Dubik D., Alimonti J., Brown T., Israels S., et al. BNIP3 and genetic control of necrosis-like cell death through the mitochondrial permeability transition pore. Mol Cell Biol 2000, 20:5454-5468.
Pattingre S., Tassa A., Qu X., Garuti R., Liang X.H., Mizushima N., et al. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 2005, 122:927-939.
Burton T.R., Eisenstat D.D., Gibson S.B. BNIP3 (Bcl-2 19kDa interacting protein) acts as transcriptional repressor of apoptosis-inducing factor expression preventing cell death in human malignant gliomas. J Neurosci 2009, 29:4189-4199.