Abstract
Current cancer therapies including cytotoxic chemotherapy, radiation and hyperthermic therapy induce acute proteotoxic stress in tumour cells. A major challenge to cancer therapeutic efficacy is the recurrence of therapy-resistant tumours and how to overcome their emergence. The current study examines the concept that tumour cell exposure to acute proteotoxic stress results in the acquisition of a more advanced and aggressive cancer cell phenotype. Specifically, we determined whether heat stress resulted in an epithelial-to-mesenchymal transition (EMT) and/or the enhancement of cell migration, components of an advanced and therapeutically resistant cancer phenotype. We identified that heat stress enhanced cell migration in both the lung A549, and breast MDA-MB-468 human adenocarcinoma cell lines, with A549 cells also undergoing a partial EMT. Moreover, in an in vivo model of thermally ablated liver metastases of the mouse colorectal MoCR cell line, immunohistological analysis of classical EMT markers demonstrated a shift to a more mesenchymal phenotype in the surviving tumour fraction, further demonstrating that thermal stress can induce epithelial plasticity. To identify a mechanism by which thermal stress modulates epithelial plasticity, we examined whether the major transcriptional regulator of the heat shock response, heat shock factor 1 (HSF1), was a required component. Knockdown of HSF1 in the A549 model did not prevent the associated morphological changes or enhanced migratory profile of heat stressed cells. Therefore, this study provides evidence that heat stress significantly impacts upon cancer cell epithelial plasticity and the migratory phenotype independent of HSF1. These findings further our understanding of novel biological downstream effects of heat stress and their potential independence from the classical heat shock pathway.
Similar content being viewed by others
References
Anckar J, Sistonen L (2011) Regulation of HSF1 function in the heat stress response: implications in aging and disease. Annu Rev Biochem 80:1089–1115. doi:10.1146/annurev-biochem-060809-095203
Baritaki S, Chapman A, Yeung K, Spandidos DA, Palladino M, Bonavida B (2009) Inhibition of epithelial to mesenchymal transition in metastatic prostate cancer cells by the novel proteasome inhibitor, NPI-0052: pivotal roles of snail repression and RKIP induction. Oncogene 28(40):3573–3585. doi:10.1038/onc.2009.214
Blagosklonny MV (2005a) Carcinogenesis, cancer therapy and chemoprevention. Cell Death Differ 12:592–602
Blagosklonny MV (2005b) Why therapeutic response may not prolong the life of a cancer patient. Cell Cycle 4(12):1693–1698
Cannito S, Novo E, Compagnone A, Valfre di Bonzo L, Busletta C, Zmara E, Paternostro C, Povero D, Bandino A, Bozzo F, Cravanzola C, Bravoco V, Colombatto S, Parola M (2008) Redox mechanisms switch on hypoxia-dependent epithelial-mesenchymal transition in cancer cells. Carcinogenesis 29(12):2267–2278
Chakraborty PK, Scharner B, Jurasovic J, Messner B, Bernhard D, Thevenod F (2010) Chronic cadmium exposure induces transcriptional activation of the Wnt pathway and upregulation of epithelial-to-mesenchymal transition markers in mouse kidney. Toxicol Lett 198(1):69–76
Ciocca DR, Calderwood SK (2005) Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. Cell Stress Chaperones 10(2):86–103
Condeelis J, Singer RH, Segall JE (2005) The great escape: when cancer cells hijack the genes for chemotaxis and motility. The Ann Rev Cell Dev Biol 21:695–718. doi:10.1146/
Dai C, Whitesell L, Rogers AB, Lindquist S (2007) Heat shock factor 1 is a powerful multifaceted modifier of carcinogenesis. Cell 130(6):1005–1018. doi:10.1016/j.cell.2007.07.020
Dai C, Dai S, Cao J (2012) Proteotoxic stress of cancer: implication of the heat-shock response in oncogenesis. J Cell Physiol 227(8):2982–2987. doi:10.1002/jcp.24017
Dou QP, Li B (1999) Proteasome inhibitors as potential novel anticancer agents. Drug Resist Updates 2:215–223
Dubois M, Bensaude O (1993) MAP kinase activation during heat shock in quiescent and exponentially growing mammalian cells. FEBS Lett 324(2):191–195
Ebos JM, Lee CR, Cruz-Munoz W, Bjarnason GA, Christensen JG, Kerbel RS (2009) Accelerated metastasis after short-term treatment with a potent inhibitor of tumor angiogenesis. Cancer Cell 15(3):232–239. doi:10.1016/j.ccr.2009.01.021
Fifis T, Malcontenti-Wilson C, Amijoyo J, Anggono B, Muralidharan V, Nikfarjam M, Christophi C (2011) Changes in growth factor levels after thermal ablation in a murine model of colorectal liver metastases. HPB: Offic J Intl Hepatol Pancreatol Biliary Assoc 13(4):246–255. doi:10.1111/j.1477-2574.2010.00278.x
Forsyth CB, Tang Y, Shaikh M, Zhang L, Keshavarzian A (2010) Alcohol stimulates activation of Snail, epidermal growth factor receptor signaling, and biomarkers of epithelial-mesenchymal transition in colon and breast cancer cells. Alcohol Clin Exp Res 34(1):19–31. doi:10.1111/j.1530-0277.2009.01061.x
Fribley A, Zeng Q, Wang CY (2004) Proteasome inhibitor PS-341 induces apoptosis through induction of endoplasmic reticulum stress-reactive oxygen species in head and neck squamous cell carcinoma cells. Mol Cell Biol 24(22):9695–9704. doi:10.1128/MCB.24.22.9695-9704.2004
Garcia MP, Cavalheiro JR, Fernandes MH (2012) Acute and long-term effects of hyperthermia in b16-f10 melanoma cells. PLoS One 7(4):e35489. doi:10.1371/journal.pone.0035489
Guettouche T, Boellmann F, Lane WS, Voellmy R (2005) Analysis of phosphorylation of human heat shock factor 1 in cells experiencing a stress. BMC Biochem 6:4. doi:10.1186/1471-2091-6-4
Hayashida N, Fujimoto M, Tan K, Prakasam R, Shinkawa T, Li L, Ichikawa H, Takii R, Nakai A (2010) Heat shock factor 1 ameliorates proteotoxicity in cooperation with the transcription factor NFAT. EMBO J 29(20):3459–3469. doi:10.1038/emboj.2010.225
Heldens L, Hensen SM, Onnekink C, van Genesen ST, Dirks RP, Lubsen NH (2011) An atypical unfolded protein response in heat shocked cells. PLoS One 6(8):e23512. doi:10.1371/journal.pone.0023512
Hildebrandt B, Wust P, Ahlers O, Dieing A, Sreenivasa G, Kerner T, Felix R, Riess H (2002) The cellular and molecular basis of hyperthermia. Crit Rev Oncol/Hematol 43:33–56
Hsu Y, Yu H, Lin H, Wu K, Yang R, Kuo P (2011) Heat shock induces apoptosis through reactive oxygen species involving mitochondrial and death receptor pathways in corneal cells. Exp Eye Res 93(4):405–412
Huber MA, Kraut N, Beug H (2005) Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 17(5):548–558. doi:10.1016/j.ceb.2005.08.001
Huff CA, Matsui W, Smith BD, Jones RJ (2006) The paradox of response and survival in cancer therapeutics. Blood 107(2):431–434. doi:10.1182/blood-2005-06-2517
Ianaro A, Ialenti A, Maffia P, Pisano B, Di Rosa M (2001) HSF1/hsp72 pathway as an endogenous anti-inflammatory system. FEBS Lett 499(3):239–244
Jolly C, Morimoto RI (2000) Role of the heat shock response and molecular chaperones in oncogenesis and cell death. J Natl Cancer Inst 19:1564–1572
Jozwiak Z, Leyko W (1992) Role of the membrane components in thermal injury of cells and development of thermotolerance. Int J Radiat Biol 62(6):743–756
Kasai H, Allen JT, Mason RM, Kamimura T, Zhang Z (2005) TGF-beta1 induces human alveolar epithelial to mesenchymal cell transition (EMT). Respir Res 6:56. doi:10.1186/1465-9921-6-56
Khalil AA, Kabapy NF, Deraz SF, Smith C (2011) Heat shock proteins in oncology: diagnostic biomarkers or therapeutic targets? Biochim Biophys Acta 1816(2):89–104. doi:10.1016/j.bbcan.2011.05.001
Kouspou MM, Price JT (2011) Analysis of cellular migration using a two-chamber methodology. Methods Mol Biol 787:303–317
Li QQ, Xu JD, Wang WJ, Cao XX, Chen Q, Tang F, Chen ZQ, Liu XP, Xu ZD (2009) Twist1-mediated adriamycin-induced epithelial-mesenchymal transition relates to multidrug resistance and invasive potential in breast cancer cells. Clin Cancer Res 15(8):2657–2665. doi:10.1158/1078-0432.CCR-08-2372
Lin RZ, Hu Z-W, Chin JH, Hoffman BB (1997) Heat shock activates c-Src tyrosine kinases and phosphatidylinositol 3-kinase in NIH3T3 fibroblasts. J Biol Chem 272(49):31196–31202
Lo HW, Hsu SC, Xia W, Cao X, Shih JY, Wei Y, Abbruzzese JL, Hortobagyi GN, Hung MC (2007) Epidermal growth factor receptor cooperates with signal transducer and activator of transcription 3 to induce epithelial-mesenchymal transition in cancer cells via up-regulation of TWIST gene expression. Cancer Res 67(19):9066–9076. doi:10.1158/0008-5472.CAN-07-0575
Maity TK, Henry MM, Tulapurkar ME, Shah NG, Hasday JD, Singh IS (2011) Distinct, gene-specific effect of heat shock on heat shock factor-1 recruitment and gene expression of CXC chemokine genes. Cytokine 54(1):61–67. doi:10.1016/j.cyto.2010.12.017
Mak P, Leav I, Pursell B, Bae D, Yang X, Taglienti CA, Gouvin LM, Sharma VM, Mercurio AM (2010) ERbeta impedes prostate cancer EMT by destabilizing HIF-1alpha and inhibiting VEGF-mediated snail nuclear localization: implications for Gleason grading. Cancer Cell 17(4):319–332. doi:10.1016/j.ccr.2010.02.030
McMillan RD, Xiao X, Shao L, Graves K, Benjamin IJ (1998) Targeted disruption of heat shock transcription factor 1 abolishes thermotolerance and protection against heat-inducible apoptosis. J Biol Chem 273(13):7523–7528
Nadeau SI, Landry J (2007) Mechanisms of activation and regulation of the heat shock-sensitive signaling pathways. Adv Exp Med Biol 594:100–113
Neznanov N, Komarov AP, Neznanova L, Stanhope-Baker P, Gudkov AV (2011) Proteotoxic stress targeted therapy (PSTT): induction of protein misfolding enhances the antitumor effect of the proteasome inhibitor bortezomib. Oncotarget 2(3):209–221
Nikfarjam M, Muralidharan V, Su K, Malcontenti-Wilson C, Christophi C (2005) Patterns of heat shock protein (HSP70) expression and Kupffer cell activity following thermal ablation of liver and colorectal liver metastases. Int J Hyperth 21(4):319–332. doi:10.1080/02656730500133736
O’Callaghan-Sunol C, Sherman MY (2006) Heat shock transcription factor (HSF1) plays a critical role in cell migration via maintaining MAP kinase signaling. Cell Cycle 5(13):1431–1437
Oliveira-Filho RS, Bevilacqua RG, Chammas R (1997) Hyperthermia increases the metastatic potential of murine melanoma. Braz J Med Biol Res 30:941–945
Paez-Ribes M, Allen E, Hudock J, Takeda T, Okuyama H, Vinals F, Inoue M, Bergers G, Hanahan D, Casanovas O (2009) Antiangiogenic therapy elicits malignant progression of tumors to increased local invasion and distant metastasis. Cancer Cell 15(3):220–231. doi:10.1016/j.ccr.2009.01.027
Page TJ, Sikder D, Yang L, Pluta L, Wolfinger RD, Kodadek T, Thomas RS (2006) Genome-wide analysis of human HSF1 signaling reveals a transcriptional program linked to cellular adaptation and survival. Mol Biosyst 2(12):627–639. doi:10.1039/b606129j
Price JT, Quinn JM, Sims NA, Vieusseux J, Waldeck K, Docherty SE, Myers D, Nakamura A, Waltham MC, Gillespie MT, Thompson EW (2005) The heat shock protein 90 inhibitor, 17-allylamino-17-demethoxygeldanamycin, enhances osteoclast formation and potentiates bone metastasis of a human breast cancer cell line. Cancer Res 65(11):4929–4938. doi:10.1158/0008-5472.CAN-04-4458
Robson EJD, Khaled WT, Abell K, Watson CJ (2006) Epithelial-to-mesenchymal transition confers resistance to apoptosis in three murine mammary epithelial cell lines. Differentiation 74(5):254–264
Ruijter JM, Ramakers C, Hoogaars WM, Karlen Y, Bakker O, van den Hoff MJ, Moorman AF (2009) Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res 37(6):e45. doi:10.1093/nar/gkp045
Rylander MN, Feng Y, Bass J, Diller KR (2005) Thermally induced injury and heat-shock protein expression in cells and tissues. Ann N Y Acad Sci 1066:222–242. doi:10.1196/annals.1363.009
Saitoh M, Shirakihara T, Miyazono K (2009) Regulation of the stability of cell surface E-cadherin by the proteasome. Biochem Biophys Res Commun 381(4):560–565. doi:10.1016/j.bbrc.2009.02.098
Samali A, Cotter TG (1996) Heat shock proteins increase resistance to apoptosis. Exper Cell Res 223:163–170
Santagata S, Hu R, Lin NU, Mendillo ML, Collins LC, Hankinson SE, Schnitt SJ, Whitesell L, Tamimi RM, Lindquist S, Ince TA (2011) High levels of nuclear heat-shock factor 1 (HSF1) are associated with poor prognosis in breast cancer. PNAS 108(45):18378–18383
Schmalhofer O, Brabletz S, Brabletz T (2009) E-cadherin, beta-catenin, and ZEB1 in malignant progression of cancer. Cancer Metastasis Rev 28(1–2):151–166. doi:10.1007/s10555-008-9179-y
Singh IS, Gupta A, Nagarsekar A, Cooper Z, Manka C, Hester L, Benjamin IJ, He JR, Hasday JD (2008) Heat shock co-activates interleukin-8 transcription. Am J Respir Cell Mol Biol 39(2):235–242. doi:10.1165/rcmb.2007-0294OC
Spaderna S, Schmalhofer O, Wahlbuhl M, Dimmler A, Bauer K, Sultan A, Hlubek F, Jung A, Strand D, Eger A, Kirchner T, Behrens J, Brabletz T (2008) The transcriptional repressor ZEB1 promotes metastasis and loss of cell polarity in cancer. Cancer Res 68(2):537–544. doi:10.1158/0008-5472.CAN-07-5682
Tamminen JA, Myllärniemi M, Hyytiäinen M, Keski-Oja J, Koli K (2012) Asbestos exposure induces alveolar epithelial cell plasticity through MAPK/Erk signaling. J Cell Biochem:n/a-n/a. doi:10.1002/jcb.24094
Thiery JP, Sleeman JP (2006) Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 7(2):131–142. doi:10.1038/nrm1835
Tiwari N, Gheldof A, Tatari M, Christofori G (2012) EMT as the ultimate survival mechanism of cancer cells. Semin Cancer Biol. doi:10.1016/j.semcancer.2012.02.013
Trinklein ND, Murray JI, Hartman SJ, Botstein D, Myers RM (2004) The role of heat shock transcription factor 1 in the genome-wide regulation of the mammalian heat shock response. Mol Biol Cell 15(3):1254–1261. doi:10.1091/mbc.E03-10-0738
Vilaboa NE, Galan A, Troyano A, de Blas E, Aller P (2000) Regulation of multidrug resistance 1 (MDR1)/P-glycoprotein gene expression and activity by heat-shock transcription factor 1 (HSF1). J Biol Chem 275(32):24970–24976. doi:10.1074/jbc.M909136199
Wang W, Goswami S, Sahai E, Wyckoff JB, Segall JE, Condeelis JS (2005) Tumor cells caught in the act of invading: their strategy for enhanced cell motility. Trends Cell Biol 15(3):138–145. doi:10.1016/j.tcb.2005.01.003
Wei L, Liu TT, Wang HH, Hong HM, Yu AL, Feng HP, Chang WW (2011) Hsp27 participates in the maintenance of breast cancer stem cells through regulation of epithelial-mesenchymal transition and nuclear factor-kappaB. Breast Cancer Res: BCR 13(5):R101. doi:10.1186/bcr3042
Whitesell L, Lindquist S (2009) Inhibiting the transcription factor HSF1 as an anticancer strategy. Expert Opin Ther Targets 13(4):469–478
Wolf F, Li W, Li F, Li C (2011) Non-invasive, quantitiative monitoring of hyperthermia-induced EGFR activation in xenograft tumours. Int J Hyperth 27(5):427–434
Workman P, Davies FE (2011) A stressful life (or death): combinatorial proteotoxic approaches to cancer0selective therapeutic vulnerability. Oncotarget 2(4):277–280
Xiao X, Zuo X, Davis AA, McMillan DR, Curry BB, Richardson JA, Benjamin IJ (1999) HSF1 is required for extra-embryonic development, postnatal growth and protection during inflammatory responses in mice. EMBO J 18(21):5943–5952
Xie K, Huang S (2003) Regulation of cancer metastasis by stress pathways. Clin Exp Metastasis 20:31–43
Yang Y, Kitagaki J, Wang H, Hou DX, Perantoni AO (2009) Targeting the ubiquitin-proteasome system for cancer therapy. Cancer Sci 100(1):24–28. doi:10.1111/j.1349-7006.2008.01013.x
Zaarur N, Gabai VL, Porco JA Jr, Calderwood S, Sherman MY (2006) Targeting heat shock response to sensitize cancer cells to proteasome and Hsp90 inhibitors. Cancer Res 66(3):1783–1791. doi:10.1158/0008-5472.CAN-05-3692
Zeisberg M, Neilson EG (2009) Biomarkers for epithelial-mesenchymal transitions. J Clin Invest 119(6):1429–1437. doi:10.1172/JCI36183
Zhong Q, Zhou B, Ann DK, Minoo P, Liu Y, Banfalvi A, Krishnaveni MS, Dubourd M, Demaio L, Willis BC, Kim KJ, duBois RM, Crandall ED, Beers MF, Borok Z (2011) Role of endoplasmic reticulum stress in epithelial-mesenchymal transition of alveolar epithelial cells: effects of misfolded surfactant protein. Am J Respir Cell Molr Biol 45(3):498–509. doi:10.1165/rcmb.2010-0347OC
Acknowledgments
The authors would like to acknowledge the staff at Monash Micro Imaging and Irene Hatzinisiriou (Monash University) for support in microscopy; staff at Flowcore (Monash University) for FACS support; Brendan Wilding (Monash University) for assistance with software analysis and Ashleigh Unsworth (Monash University) for critical reading of the manuscript. This work was supported by Cancer Council Victoria grant-in-aid no. 545969, National Health and Medical Research Council of Australia R Douglas Wright fellowship no. 395525 (JTP), Australian Postgraduate Award (BJL).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lang, B.J., Nguyen, L., Nguyen, H.C. et al. Heat stress induces epithelial plasticity and cell migration independent of heat shock factor 1. Cell Stress and Chaperones 17, 765–778 (2012). https://doi.org/10.1007/s12192-012-0349-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12192-012-0349-z