Abstract
In human glioma tumours, heme oxygenase-1 (HO-1) is overexpressed when compared with normal brain tissues and during oligodendroglioma progression. However, the molecular mechanisms mediated by HO-1 to promote glioblastoma remain unknown. We therefore aimed at investigating the effect of HO-1 expression and its selective enzymatic inhibition in two different cell lines (i.e. A172 and U87-MG). HO-1 was induced by hemin treatment (10 μM), and VP13/47 (100 μM) was used as a specific non-competitive inhibitor of HO-1 activity. Cell proliferation was measured by cell index measurement (xCelligence technology) and clonogenic assay, whereas cell migration was assessed by wound healing assay. Carbon monoxide-releasing molecules (CORMs) (i.e. CORM-3 and CORM-A1) were also used in a separate set of experiments to confirm the effect of HO-1 by-product in glioblastoma progression further. Our results were further validated using GSE4412 microarray dataset analysis and comparing biopsies overexpressing HO-1 with the rest of the cases. Our results showed that hemin was able to induce both HO-1 gene and protein expression in a cell-dependent manner being A172 more responsive to pharmacological upregulation of HO-1. Hemin, but not CORMs treatment, resulted in a significant increase of cell proliferation following 24 h of treatment as measured by increased cell index and colony formation capacity and such effect was abolished by VP13/47. Interestingly, both hemin and CORMs showed a significant effect on the wound healing assay also exhibiting cell specificity. Finally, our dataset analysis showed a positive correlation of HO-1 gene expression with ITGBI and ITGBII which are membrane receptors involved in cell adhesion, embryogenesis, tissue repair, immune response and metastatic diffusion of tumour cells. In conclusion, our data suggest that HO-1 and its by-product CO exhibit a cell-specific effect on various aspects of disease progression and are associated with a complex series of molecular mechanisms driving cell proliferation, survival and metastasis.
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References
Lefranc F, Le Rhun E, Kiss R, Weller M (2018) Glioblastoma quo vadis: will migration and invasiveness reemerge as therapeutic targets? Cancer Treat Rev 68:145–154. https://doi.org/10.1016/j.ctrv.2018.06.017
Ventura E, Weller M, Burghardt I (2017) Cutting edge: ERK1 mediates the autocrine positive feedback loop of TGF-beta and furin in glioma-initiating cells. J Immunol 198(12):4569–4574. https://doi.org/10.4049/jimmunol.1601176
Suzuki H, Aoki K, Chiba K, Sato Y, Shiozawa Y, Shiraishi Y, Shimamura T, Niida A et al (2015) Mutational landscape and clonal architecture in grade II and III gliomas. Nat Genet 47(5):458–468. https://doi.org/10.1038/ng.3273
Parsons DW, Jones S, Zhang X, Lin JC, Leary RJ, Angenendt P, Mankoo P, Carter H et al (2008) An integrated genomic analysis of human glioblastoma multiforme. Science 321(5897):1807–1812. https://doi.org/10.1126/science.1164382
Sanson M, Marie Y, Paris S, Idbaih A, Laffaire J, Ducray F, El Hallani S, Boisselier B et al (2009) Isocitrate dehydrogenase 1 codon 132 mutation is an important prognostic biomarker in gliomas. J Clin Oncol 27(25):4150–4154. https://doi.org/10.1200/JCO.2009.21.9832
Yan H, Parsons DW, Jin G, McLendon R, Rasheed BA, Yuan W, Kos I, Batinic-Haberle I et al (2009) IDH1 and IDH2 mutations in gliomas. N Engl J Med 360(8):765–773. https://doi.org/10.1056/NEJMoa0808710
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA et al (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352(10):987–996. https://doi.org/10.1056/NEJMoa043330
Perry JR, Laperriere N, O'Callaghan CJ, Brandes AA, Menten J, Phillips C, Fay M, Nishikawa R et al (2017) Short-course radiation plus temozolomide in elderly patients with glioblastoma. N Engl J Med 376(11):1027–1037. https://doi.org/10.1056/NEJMoa1611977
Weller M, van den Bent M, Tonn JC, Stupp R, Preusser M, Cohen-Jonathan-Moyal E, Henriksson R, Le Rhun E, Balana C, Chinot O, Bendszus M, Reijneveld JC, Dhermain F, French P, Marosi C, Watts C, Oberg I, Pilkington G, Baumert BG, Taphoorn MJB, Hegi M, Westphal M, Reifenberger G, Soffietti R, Wick W, European Association for Neuro-Oncology Task Force on G (2017) European Association for Neuro-Oncology (EANO) guideline on the diagnosis and treatment of adult astrocytic and oligodendroglial gliomas. Lancet Oncol 18 (6):e315-e329. doi:https://doi.org/10.1016/S1470-2045(17)30194-8
Haas-Kogan DA, Prados MD, Tihan T, Eberhard DA, Jelluma N, Arvold ND, Baumber R, Lamborn KR et al (2005) Epidermal growth factor receptor, protein kinase B/Akt, and glioma response to erlotinib. J Natl Cancer Inst 97(12):880–887. https://doi.org/10.1093/jnci/dji161
Ahmad Z, Salim M, Maines MD (2002) Human biliverdin reductase is a leucine zipper-like DNA-binding protein and functions in transcriptional activation of heme oxygenase-1 by oxidative stress. J Biol Chem 277(11):9226–9232. https://doi.org/10.1074/jbc.M108239200
Tenhunen R, Marver H, Pimstone NR, Trager WF, Cooper DY, Schmid R (1972) Enzymatic degradation of heme. Oxygenative cleavage requiring cytochrome P-450. Biochemistry 11(9):1716–1720. https://doi.org/10.1021/bi00759a029
Abraham NG, Feldman E, Falck JR, Lutton JD, Schwartzman ML (1991) Modulation of erythropoiesis by novel human bone marrow cytochrome P450-dependent metabolites of arachidonic acid. Blood 78(6):1461–1466
Kapitulnik J, Maines MD (2009) Pleiotropic functions of biliverdin reductase: cellular signaling and generation of cytoprotective and cytotoxic bilirubin. Trends Pharmacol Sci 30(3):129–137. https://doi.org/10.1016/j.tips.2008.12.003
Keyse SM, Tyrrell RM (1989) Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci U S A 86(1):99–103. https://doi.org/10.1073/pnas.86.1.99
Foresti R, Clark JE, Green CJ, Motterlini R (1997) Thiol compounds interact with nitric oxide in regulating heme oxygenase-1 induction in endothelial cells. Involvement of superoxide and peroxynitrite anions. J Biol Chem 272(29):18411–18417. https://doi.org/10.1074/jbc.272.29.18411
Grochot-Przeczek A, Dulak J, Jozkowicz A (2012) Haem oxygenase-1: non-canonical roles in physiology and pathology. Clin Sci (Lond) 122(3):93–103. https://doi.org/10.1042/CS20110147
Kim HP, Wang X, Galbiati F, Ryter SW, Choi AM (2004) Caveolae compartmentalization of heme oxygenase-1 in endothelial cells. FASEB J 18(10):1080–1089. https://doi.org/10.1096/fj.03-1391com
Converso DP, Taille C, Carreras MC, Jaitovich A, Poderoso JJ, Boczkowski J (2006) HO-1 is located in liver mitochondria and modulates mitochondrial heme content and metabolism. FASEB J 20(8):1236–1238. https://doi.org/10.1096/fj.05-4204fje
Zijlstra GS, Brandsma CA, Harpe MF, Van Dam GM, Slebos DJ, Kerstjens HA, De Boer AH, Frijlink HW (2007) Dry powder inhalation of hemin to induce heme oxygenase expression in the lung. Eur J Pharm Biopharm 67(3):667–675. https://doi.org/10.1016/j.ejpb.2007.03.021
Vanella L, Barbagallo I, Tibullo D, Forte S, Zappala A, Li Volti G (2016) The non-canonical functions of the heme oxygenases. Oncotarget 7(42):69075–69086. https://doi.org/10.18632/oncotarget.11923
Biswas C, Shah N, Muthu M, La P, Fernando AP, Sengupta S, Yang G, Dennery PA (2014) Nuclear heme oxygenase-1 (HO-1) modulates subcellular distribution and activation of Nrf2, impacting metabolic and anti-oxidant defenses. J Biol Chem 289(39):26882–26894. https://doi.org/10.1074/jbc.M114.567685
Loboda A, Jozkowicz A, Dulak J (2015) HO-1/CO system in tumor growth, angiogenesis and metabolism - targeting HO-1 as an anti-tumor therapy. Vasc Pharmacol 74:11–22. https://doi.org/10.1016/j.vph.2015.09.004
Schacter BA, Kurz P (1986) Alterations in microsomal drug metabolism and heme oxygenase activity in isolated hepatic parenchymal and sinusoidal cells in Murphy-Sturm lymphosarcoma-bearing rats. Clin Invest Med 9(3):150–155
Goodman AI, Choudhury M, da Silva JL, Schwartzman ML, Abraham NG (1997) Overexpression of the heme oxygenase gene in renal cell carcinoma. Proc Soc Exp Biol Med 214(1):54–61. https://doi.org/10.3181/00379727-214-44069
Maines MD, Abrahamsson PA (1996) Expression of heme oxygenase-1 (HSP32) in human prostate: normal, hyperplastic, and tumor tissue distribution. Urology 47(5):727–733. https://doi.org/10.1016/s0090-4295(96)00010-6
Was H, Dulak J, Jozkowicz A (2010) Heme oxygenase-1 in tumor biology and therapy. Curr Drug Targets 11(12):1551–1570
Tibullo D, Barbagallo I, Giallongo C, Vanella L, Conticello C, Romano A, Saccone S, Godos J et al (2016) Heme oxygenase-1 nuclear translocation regulates bortezomibinduced cytotoxicity and mediates genomic instability in myeloma cells. Oncotarget 7(20):28868–28880. https://doi.org/10.18632/oncotarget.7563
Fang J, Greish K, Qin H, Liao L, Nakamura H, Takeya M, Maeda H (2012) HSP32 (HO-1) inhibitor, copoly(styrene-maleic acid)-zinc protoporphyrin IX, a water-soluble micelle as anticancer agent: In vitro and in vivo anticancer effect. Eur J Pharm Biopharm 81(3):540–547. https://doi.org/10.1016/j.ejpb.2012.04.016
Furfaro AL, Macay JR, Marengo B, Nitti M, Parodi A, Fenoglio D, Marinari UM, Pronzato MA et al (2012) Resistance of neuroblastoma GI-ME-N cell line to glutathione depletion involves Nrf2 and heme oxygenase-1. Free Radic Biol Med 52(2):488–496. https://doi.org/10.1016/j.freeradbiomed.2011.11.007
Tan Q, Wang H, Hu Y, Hu M, Li X, Aodengqimuge MY, Wei C, Song L (2015) Src/STAT3-dependent heme oxygenase-1 induction mediates chemoresistance of breast cancer cells to doxorubicin by promoting autophagy. Cancer Sci 106(8):1023–1032. https://doi.org/10.1111/cas.12712
Lv X, Song DM, Niu YH, Wang BS (2016) Inhibition of heme oxygenase-1 enhances the chemosensitivity of laryngeal squamous cell cancer Hep-2 cells to cisplatin. Apoptosis 21(4):489–501. https://doi.org/10.1007/s10495-016-1216-7
Jeon WK, Hong HY, Seo WC, Lim KH, Lee HY, Kim WJ, Song SY, Kim BC (2012) Smad7 sensitizes A549 lung cancer cells to cisplatin-induced apoptosis through heme oxygenase-1 inhibition. Biochem Biophys Res Commun 420(2):288–292. https://doi.org/10.1016/j.bbrc.2012.02.151
Yin Y, Liu Q, Wang B, Chen G, Xu L, Zhou H (2012) Expression and function of heme oxygenase-1 in human gastric cancer. Exp Biol Med (Maywood) 237(4):362–371. https://doi.org/10.1258/ebm.2011.011193
Liu YS, Li HS, Qi DF, Zhang J, Jiang XC, Shi K, Zhang XJ, Zhang XH (2014) Zinc protoporphyrin IX enhances chemotherapeutic response of hepatoma cells to cisplatin. World J Gastroenterol 20(26):8572–8582. https://doi.org/10.3748/wjg.v20.i26.8572
Liu Y, Liang Y, Zheng T, Yang G, Zhang X, Sun Z, Shi C, Zhao S (2011) Inhibition of heme oxygenase-1 enhances anti-cancer effects of arsenic trioxide on glioma cells. J Neuro-Oncol 104(2):449–458. https://doi.org/10.1007/s11060-010-0513-1
Fest S, Soldati R, Christiansen NM, Zenclussen ML, Kilz J, Berger E, Starke S, Lode HN et al (2016) Targeting of heme oxygenase-1 as a novel immune regulator of neuroblastoma. Int J Cancer 138(8):2030–2042. https://doi.org/10.1002/ijc.29933
Gandini NA, Fermento ME, Salomon DG, Obiol DJ, Andres NC, Zenklusen JC, Arevalo J, Blasco J et al (2014) Heme oxygenase-1 expression in human gliomas and its correlation with poor prognosis in patients with astrocytoma. Tumour Biol 35(3):2803–2815. https://doi.org/10.1007/s13277-013-1373-z
Freije WA, Castro-Vargas FE, Fang Z, Horvath S, Cloughesy T, Liau LM, Mischel PS, Nelson SF (2004) Gene expression profiling of gliomas strongly predicts survival. Cancer Res 64(18):6503–6510
Griesinger AM, Birks DK, Donson AM, Amani V, Hoffman LM, Waziri A, Wang M, Handler MH et al (2013) Characterization of distinct immunophenotypes across pediatric brain tumor types. J Immunol 191(9):4880–4888
Di Rosa M, Sanfilippo C, Libra M, Musumeci G, Malaguarnera L (2015) Different pediatric brain tumors are associated with different gene expression profiling. Acta Histochem 117(4–5):477–485
Warde-Farley D, Donaldson SL, Comes O, Zuberi K, Badrawi R, Chao P, Franz M, Grouios C, Kazi F, Lopes CT, Maitland A, Mostafavi S, Montojo J, Shao Q, Wright G, Bader GD, Morris Q (2010) The GeneMANIA prediction server: biological network integration for gene prioritization and predicting gene function Nucleic Acids Res 38 (Web Server issue):W214-220
Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, Simonovic M, Doncheva NT et al (2019) STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res 47(D1):D607–D613
Li Volti G, Wang J, Traganos F, Kappas A, Abraham NG (2002) Differential effect of heme oxygenase-1 in endothelial and smooth muscle cell cycle progression. Biochem Biophys Res Commun 296(5):1077–1082. https://doi.org/10.1016/s0006-291x(02)02054-5
Li Volti G, Sacerdoti D, Sangras B, Vanella A, Mezentsev A, Scapagnini G, Falck JR, Abraham NG (2005) Carbon monoxide signaling in promoting angiogenesis in human microvessel endothelial cells. Antioxid Redox Signal 7(5–6):704–710. https://doi.org/10.1089/ars.2005.7.704
Bertozzi G, Sessa F, Albano GD, Sani G, Maglietta F, Roshan MHK, Volti GL, Bernardini R et al (2018) The role of anabolic androgenic steroids in disruption of the physiological function in discrete areas of the central nervous system. Mol Neurobiol 55(7):5548–5556. https://doi.org/10.1007/s12035-017-0774-1
Long J, Manchandia T, Ban K, Gao S, Miller C, Chandra J (2007) Adaphostin cytoxicity in glioblastoma cells is ROS-dependent and is accompanied by upregulation of heme oxygenase-1. Cancer Chemother Pharmacol 59(4):527–535. https://doi.org/10.1007/s00280-006-0295-5
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100(1):57–70. https://doi.org/10.1016/s0092-8674(00)81683-9
Jozkowicz A, Was H, Dulak J (2007) Heme oxygenase-1 in tumors: is it a false friend? Antioxid Redox Signal 9(12):2099–2117. https://doi.org/10.1089/ars.2007.1659
Gueron G, De Siervi A, Ferrando M, Salierno M, De Luca P, Elguero B, Meiss R, Navone N et al (2009) Critical role of endogenous heme oxygenase 1 as a tuner of the invasive potential of prostate cancer cells. Mol Cancer Res 7(11):1745–1755. https://doi.org/10.1158/1541-7786.MCR-08-0325
Ferrando M, Gueron G, Elguero B, Giudice J, Salles A, Leskow FC, Jares-Erijman EA, Colombo L et al (2011) Heme oxygenase 1 (HO-1) challenges the angiogenic switch in prostate cancer. Angiogenesis 14(4):467–479. https://doi.org/10.1007/s10456-011-9230-4
Miyata Y, Kanda S, Mitsunari K, Asai A, Sakai H (2014) Heme oxygenase-1 expression is associated with tumor aggressiveness and outcomes in patients with bladder cancer: a correlation with smoking intensity. Transl Res 164(6):468–476. https://doi.org/10.1016/j.trsl.2014.06.010
Li C, Wu H, Wang S, Zhu J (2016) Expression and correlation of NRF2, KEAP1, NQO-1 and HO-1 in advanced squamous cell carcinoma of the larynx and their association with clinicopathologic features. Mol Med Rep 14(6):5171–5179. https://doi.org/10.3892/mmr.2016.5913
Sebastian VP, Salazar GA, Coronado-Arrazola I, Schultz BM, Vallejos OP, Berkowitz L, Alvarez-Lobos MM, Riedel CA et al (2018) Heme oxygenase-1 as a modulator of intestinal inflammation development and progression. Front Immunol 9:1956. https://doi.org/10.3389/fimmu.2018.01956
Li Q, Li C, Chen J, Liu P, Cui Y, Zhou X, Li H, Zu X (2018) High expression of long noncoding RNA NORAD indicates a poor prognosis and promotes clinical progression and metastasis in bladder cancer. Urol Oncol 36(6):310 e315–310 e322. https://doi.org/10.1016/j.urolonc.2018.02.019
Naudin C, Hattabi A, Michelet F, Miri-Nezhad A, Benyoucef A, Pflumio F, Guillonneau F, Fichelson S et al (2017) PUMILIO/FOXP1 signaling drives expansion of hematopoietic stem/progenitor and leukemia cells. Blood 129(18):2493–2506. https://doi.org/10.1182/blood-2016-10-747436
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This work was supported by Research Funding for University of Catania, Italy (Piano per la Ricerca, FIR 2018-2020). This work was part of the PhD thesis of Dr. Carlo Castruccio Castracani (Neuroscience International PhD program).
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CCC and GLV made a substantial contribution to the concept and design, acquisition of data or analysis and interpretation of data; GLV drafted the article or revised it critically for important intellectual content; LL, AD, GL, DC, DT, VP and GLV performed in vitro experiments; MDR: analysed GEO datasets and performed statistical analysis; all the authors approved the version to be published.
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Supplementary Figure 1.
Expression levels of HO-1 in several brain tumours. Gene expression profiles were used to identify HO-1 gene that is differentially expressed between various brain tumour types. Data are expressed as z-score intensity expression levels and presented as vertical scatter dot plots and violin plots. P values <0.05 were considered to be statistically significant (*p<0.05; **p<0.005; ***p<0.0005; ****p<0.00005). (PNG 534 kb)
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Castruccio Castracani, C., Longhitano, L., Distefano, A. et al. Heme Oxygenase-1 and Carbon Monoxide Regulate Growth and Progression in Glioblastoma Cells. Mol Neurobiol 57, 2436–2446 (2020). https://doi.org/10.1007/s12035-020-01869-7
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DOI: https://doi.org/10.1007/s12035-020-01869-7