Abstract
Hypoxia-inducible factor-1α (HIF-1α) is a principal regulator of angiogenesis and other cellular responses to hypoxic stress in both normal and tumor cells. To identify novel mechanisms that regulate expression of HIF-1α, we designed a genome-wide screen for expressed sequence tags (ESTs) that when transcribed in the antisense direction increase production of the HIF-1α target, vascular endothelial growth factor (VEGF), in human breast cancer cells. We discovered that heat shock factor (HSF) proteins 2 and 4—which previously have been implicated in the control of multiple genes that modulate cell growth and differentiation and protect against effects of environmental and cellular stresses—function together to maintain a steady state level of HIF-1α transcription and VEGF production in these cells. We show both HSFs bind to discontinuous heat shock element (HSE) sequences we identified in the HIF-1α promoter region and that downregulation of either HSF activates transcription of HIF-1α. We further demonstrate that HSF2 and HSF4 displace each other from HSF/HSE complexes in the HIF-1α promoter so that HIF-1α transcription is also activated by overexpression of either HSFs. These results argue that HSF2 and HSF4 regulate transcription of HIF-1α and that a critical balance between these HSF is required to maintain HIF-α expression in a repressed state. Our findings reveal a previously unsuspected role for HSFs in control of VEGF and other genes activated by canonical HIF-1α-mediated signaling.
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References
Akerfelt M, Henriksson E, Laiho A, Vihervaara A, Rautoma K, Kotaja N et al. (2008). Promoter ChIP-chip analysis in mouse testis reveals Y chromosome occupancy by HSF2. Proc Natl Acad Sci USA 105: 11224–11229.
Akerfelt M, Trouillet D, Mezger V, Sistonen L . (2007). Heat shock factors at a crossroad between stress and development. Ann NY Acad Sci 1113: 15–27.
Bos R, Zhong H, Hanrahan CF, Mommers EC, Semenza GL, Pinedo HM et al. (2001). Levels of hypoxia-inducible factor-1 alpha during breast carcinogenesis. J Natl Cancer Inst 93: 309–314.
Brahimi-Horn MC, Pouyssegur J . (2007). Harnessing the hypoxia-inducible factor in cancer and ischemic disease. Biochem Pharmacol 73: 450–457.
Busca R, Berra E, Gaggioli C, Khaled M, Bille K, Marchetti B et al. (2005). Hypoxia-inducible factor 1{alpha} is a new target of microphthalmia-. J Cell Biol 170: 49–59.
Chang Y, Ostling P, Akerfelt M, Trouillet D, Rallu M, Gitton Y et al. (2006). Role of heat-shock factor 2 in cerebral cortex formation and as a regulator of p35 expression. Genes Dev 20: 836–847.
Chi NC, Karliner JS . (2004). Molecular determinants of responses to myocardial ischemia/reperfusion injury: focus on hypoxia-inducible and heat shock factors. Cardiovasc Res 61: 437–447.
Fraisl P, Mazzone M, Schmidt T, Carmeliet P . (2009). Regulation of angiogenesis by oxygen and metabolism. Dev Cell 16: 167–179.
Fujimoto M, Izu H, Seki K, Fukuda K, Nishida T, Yamada S et al. (2004). HSF4 is required for normal cell growth and differentiation during mouse lens development. EMBO J 23: 4297–4306.
Fujimoto M, Oshima K, Shinkawa T, Wang BB, Inouye S, Hayashida N et al. (2008). Analysis of HSF4 binding regions reveals its necessity for gene regulation during development and heat shock response in mouse lenses. J Biol Chem 283: 29961–29970.
Fujimoto M, Nakai A . (2010). The heat shock factor family and adaptation to proteotoxic stress. FEBS J 277: 4112–4125.
Fukumura D, Xavier R, Sugiura T, Chen Y, Park EC, Lu N et al. (1998). Tumor induction of VEGF promoter activity in stromal cells. Cell 94: 715–725.
Isaacs JS, Jung YJ, Mimnaugh EG, Martinez A, Cuttitta F, Neckers LM . (2002). Hsp90 regulates a von Hippel Lindau-independent hypoxia-inducible factor-1 alpha-degradative pathway. J Biol Chem 277: 29936–29944.
Jiang BH, Agani F, Passaniti A, Semenza GL . (1997). V-SRC induces expression of hypoxia-inducible factor 1 (HIF-1) and transcription of genes encoding vascular endothelial growth factor and enolase 1: involvement of HIF-1 in tumor progression. Cancer Res 57: 5328–5335.
Loison F, Debure L, Nizard P, le Goff P, Michel D, le Drean Y . (2006). Up-regulation of the clusterin gene after proteotoxic stress: implication of HSF1-HSF2 heterocomplexes. Biochem J 395: 223–231.
Lu Q, Wei W, Kowalski PE, Chang AC, Cohen SN . (2004). EST-based genome-wide gene inactivation identifies ARAP3 as a host protein affecting cellular susceptibility to anthrax toxin. Proc Natl Acad Sci USA 101: 17246–17251.
Mathew A, Mathur SK, Jolly C, Fox SG, Kim S, Morimoto RI . (2001). Stress-specific activation and repression of heat shock factors 1 and 2. Mol Cell Biol 21: 7163–7171.
Mazure NM, Chen EY, Laderoute KR, Giaccia AJ . (1997). Induction of vascular endothelial growth factor by hypoxia is modulated by a phosphatidylinositol 3-kinase/Akt signaling pathway in Ha-ras-transformed cells through a hypoxia inducible factor-1 transcriptional element. Blood 90: 3322–3331.
Mazure NM, Chen EY, Laderoute KR, Giaccia AJ . (1996). Oncogenic transformation and hypoxia synergistically act to modulate vascular endothelial growth factor expression. Cancer Res 56: 3436–3440.
Min JN, Zhang Y, Moskophidis D, Mivechi NF . (2004). Unique contribution of heat shock transcription factor 4 in ocular lens development and fiber cell differentiation. Genesis 40: 205–217.
Minet E, Mottet D, Michel G, Roland I, Raes M, Remacle J et al. (1999). Hypoxia-induced activation of HIF-1: role of HIF-1alpha-Hsp90 interaction. FEBS Lett 460: 251–256.
Morimoto RI . (2008). Proteotoxic stress and inducible chaperone networks in neurodegenerative disease and aging. Genes Dev 22: 1427–1438.
Murphy LA, Wilkerson DC, Hong Y, Sarge KD . (2008). PRC1 associates with the hsp70i promoter and interacts with HSF2 during mitosis. Exp Cell Res 314: 2224–2230.
Nakai A, Tanabe M, Kawazoe Y, Inazawa J, Morimoto RI, Nagata K . (1997). HSF4, a new member of the human heat shock factor family which lacks properties of a transcriptional activator. Mol Cell Biol 17: 469–481.
Nardinocchi L, Puca R, Guidolin D, Belloni AS, Bossi G, Michiels C et al. (2009). Transcriptional regulation of hypoxia-inducible factor 1alpha by HIPK2 suggests a novel mechanism to restrain tumor growth. Biochim Biophys Acta 1793: 368–377.
Ng YS, Krilleke D, Shima DT . (2006). VEGF function in vascular pathogenesis. Exp Cell Res 312: 527–537.
Olenyuk BZ, Zhang GJ, Klco JM, Nickols NG, Kaelin WG, Dervan PB . (2004). Inhibition of vascular endothelial growth factor with a sequence-specific hypoxia response element antagonist. Proc Natl Acad Sci USA 101: 16768–16773.
Ostling P, Bjork JK, Roos-Mattjus P, Mezger V, Sistonen L . (2007). Heat shock factor 2 (HSF2) contributes to inducible expression of hsp genes through interplay with HSF1. J Biol Chem 282: 7077–7086.
Page EL, Robitaille GA, Pouyssegur J, Richard DE . (2002). Induction of hypoxia-inducible factor-1alpha by transcriptional and translational mechanisms. J Biol Chem 277: 48403–48409.
Pipinikas CP, Carter ND, Corbishley CM, Fenske CD . (2008). HIF-1alpha mRNA gene expression levels in improved diagnosis of early stages of prostate cancer. Biomarkers 13: 680–691.
Pirkkala L, Nykanen P, Sistonen L . (2001). Roles of the heat shock transcription factors in regulation of the heat shock response and beyond. FASEB J 15: 1118–1131.
Pore N, Liu S, Shu HK, Li B, Haas-Kogan D, Stokoe D et al. (2004). Sp1 is involved in Akt-mediated induction of VEGF expression through an HIF-1-independent mechanism. Mol Biol Cell 15: 4841–4853.
Pouyssegur J, Dayan F, Mazure NM . (2006). Hypoxia signalling in cancer and approaches to enforce tumour regression. Nature 441: 437–443.
Sandqvist A, Bjork JK, Akerfelt M, Chitikova Z, Grichine A, Vourc'h C et al. (2009). Heterotrimerization of heat-shock factors 1 and 2 provides a transcriptional switch in response to distinct stimuli. Mol Biol Cell 20: 1340–1347.
Schafer C, Clapp P, Welsh MJ, Benndorf R, Williams JA . (1999). HSP27 expression regulates CCK-induced changes of the actin cytoskeleton in CHO-CCK-A cells. Am J Physiol 277: C1032–C1043.
Semenza GL . (2003). Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3: 721–732.
Somasundaram T, Bhat SP . (2004). Developmentally dictated expression of heat shock factors: exclusive expression of HSF4 in the postnatal lens and its specific interaction with alphaB-crystallin heat shock promoter. J Biol Chem 279: 44497–44503.
Tsuzuki Y, Fukumura D, Oosthuyse B, Koike C, Carmeliet P, Jain RK . (2000). Vascular endothelial growth factor (VEGF) modulation by targeting hypoxia-inducible factor-1alpha--> hypoxia response element--> VEGF cascade differentially regulates vascular response and growth rate in tumors. Cancer Res 60: 6248–6252.
Wartenberg M, Donmez F, Ling FC, Acker H, Hescheler J, Sauer H . (2001). Tumor-induced angiogenesis studied in confrontation cultures of multicellular tumor spheroids and embryoid bodies grown from pluripotent embryonic stem cells. FASEB J 15: 995–1005.
Xing H, Wilkerson DC, Mayhew CN, Lubert EJ, Skaggs HS, Goodson ML et al. (2005). Mechanism of hsp70i gene bookmarking. Science 307: 421–423.
Yamamoto N, Takemori Y, Sakurai M, Sugiyama K, Sakurai H . (2009). Differential recognition of heat shock elements by members of the heat shock transcription factor family. FEBS J 276: 1962–1974.
Zhang J, Goodson ML, Hong Y, Sarge KD . (2008). MEL-18 interacts with HSF2 and the SUMO E2 UBC9 to inhibit HSF2 sumoylation. J Biol Chem 283: 7464–7469.
Zhang Y, Frejtag W, Dai R, Mivechi NF . (2001). Heat shock factor-4 (HSF-4a) is a repressor of HSF-1 mediated transcription. J Cell Biochem 82: 692–703.
Acknowledgements
The study was supported by funds from the Kwoh-Ting Li Professorship to SNC, and by a grant from the National Foundation for Cancer Research (NFCR) to SNC.
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Chen, R., Liliental, J., Kowalski, P. et al. Regulation of transcription of hypoxia-inducible factor-1α (HIF-1α) by heat shock factors HSF2 and HSF4. Oncogene 30, 2570–2580 (2011). https://doi.org/10.1038/onc.2010.623
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DOI: https://doi.org/10.1038/onc.2010.623
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