Abstract
Cancer is associated with strong changes in lipid metabolism. For instance, normal cells take up fatty acids (FAs) from the circulation, while tumour cells generate their own and become dependent on de novo FA synthesis, which could provide a vulnerability to target tumour cells. Betulinic acid (BetA) is a natural compound that selectively kills tumour cells through an ill-defined mechanism that is independent of BAX and BAK, but depends on mitochondrial permeability transition-pore opening. Here we unravel this pathway and show that BetA inhibits the activity of steroyl-CoA-desaturase (SCD-1). This enzyme is overexpressed in tumour cells and critically important for cells that utilize de novo FA synthesis as it converts newly synthesized saturated FAs to unsaturated FAs. Intriguingly, we find that inhibition of SCD-1 by BetA or, alternatively, with a specific SCD-1 inhibitor directly and rapidly impacts on the saturation level of cardiolipin (CL), a mitochondrial lipid that has important structural and metabolic functions and at the same time regulates mitochondria-dependent cell death. As a result of the enhanced CL saturation mitochondria of cancer cells, but not normal cells that do not depend on de novo FA synthesis, undergo ultrastructural changes, release cytochrome c and quickly induce cell death. Importantly, addition of unsaturated FAs circumvented the need for SCD-1 activity and thereby prevented BetA-induced CL saturation and subsequent cytotoxicity, supporting the importance of this novel pathway in the cytotoxicity induced by BetA.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Swinnen JV, Brusselmans K, Verhoeven G . Increased lipogenesis in cancer cells: new players, novel targets. Curr Opin Clin Nutr Metab Care 2006; 9: 358–365.
DeBerardinis RJ, Thompson CB . Cellular metabolism and disease: what do metabolic outliers teach us? Cell 2012; 148: 1132–1144.
Santos CR, Schulze A . Lipid metabolism in cancer. FEBS J 2012; 279: 2610–2623.
Medes G, Thomas A, Weinhouse S . Metabolism of neoplastic tissue. IV. A study of lipid synthesis in neoplastic tissue slices in vitro. Cancer Res 1953; 13: 27–29.
Ookhtens M, Kannan R, Lyon I, Baker N . Liver and adipose tissue contributions to newly formed fatty acids in an ascites tumor. Am J Physiol 1984; 247: R146–R153.
Menendez JA, Lupu R . Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis. Nat Rev Cancer 2007; 7: 763–777.
Li JN, Mahmoud MA, Han WF, Ripple M, Pizer ES . Sterol regulatory element-binding protein-1 participates in the regulation of fatty acid synthase expression in colorectal neoplasia. Exp Cell Res 2000; 261: 159–165.
Swinnen JV, Vanderhoydonc F, Elgamal AA, Eelen M, Vercaeren I, Joniau S et al. Selective activation of the fatty acid synthesis pathway in human prostate cancer. Int J Cancer 2000; 88: 176–179.
Bauer DE, Hatzivassiliou G, Zhao F, Andreadis C, Thompson CB . ATP citrate lyase is an important component of cell growth and transformation. Oncogene 2005; 24: 6314–6322.
Hatzivassiliou G, Zhao F, Bauer DE, Andreadis C, Shaw AN, Dhanak D et al. ATP citrate lyase inhibition can suppress tumor cell growth. Cancer Cell 2005; 8: 311–321.
Kumar-Sinha C, Ignatoski KW, Lippman ME, Ethier SP, Chinnaiyan AM . Transcriptome analysis of HER2 reveals a molecular connection to fatty acid synthesis. Cancer Res 2003; 63: 132–139.
Scaglia N, Igal RA . Stearoyl-CoA desaturase is involved in the control of proliferation, anchorage-independent growth, and survival in human transformed cells. J Biol Chem 2005; 280: 25339–25349.
Scaglia N, Caviglia JM, Igal RA . High stearoyl-CoA desaturase protein and activity levels in simian virus 40 transformed-human lung fibroblasts. Biochim Biophys Acta 2005; 1687: 141–151.
Paton CM, Ntambi JM . Biochemical and physiological function of stearoyl-CoA desaturase. Am J Physiol Endocrinol Metab 2009; 297: E28–E37.
Scaglia N, Chisholm JW, Igal RA . Inhibition of stearoylCoA desaturase-1 inactivates acetyl-CoA carboxylase and impairs proliferation in cancer cells: role of AMPK. PLoS One 2009; 4: e6812.
Igal RA . Stearoyl-CoA desaturase-1: a novel key player in the mechanisms of cell proliferation, programmed cell death and transformation to cancer. Carcinogenesis 2010; 31: 1509–1515.
Zuco V, Supino R, Righetti SC, Cleris L, Marchesi E, Gambacorti-Passerini C et al. Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells. Cancer Lett 2002; 175: 17–25.
Rzeski W, Stepulak A, Szymanski M, Sifringer M, Kaczor J, Wejksza K et al. Betulinic acid decreases expression of bcl-2 and cyclin D1, inhibits proliferation, migration and induces apoptosis in cancer cells. Naunyn Schmiedebergs Arch Pharmacol 2006; 374: 11–20.
Mullauer FB, Kessler JH, Medema JP . Betulinic acid, a natural compound with potent anticancer effects. Anticancer Drugs 2010; 21: 215–227.
Fulda S, Friesen C, Los M, Scaffidi C, Mier W, Benedict M et al. Betulinic acid triggers CD95 (APO-1/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. Cancer Res 1997; 57: 4956–4964.
Fulda S, Susin SA, Kroemer G, Debatin KM . Molecular ordering of apoptosis induced by anticancer drugs in neuroblastoma cells. Cancer Res 1998; 58: 4453–4460.
Kessler JH, Mullauer FB, de Roo GM, Medema JP . Broad in vitro efficacy of plant-derived betulinic acid against cell lines derived from the most prevalent human cancer types. Cancer Lett 2007; 251: 132–145.
Mullauer FB, Kessler JH, Medema JP . Betulinic acid induces cytochrome c release and apoptosis in a Bax/Bak-independent, permeability transition pore dependent fashion. Apoptosis 2009; 14: 191–202.
Fry M, Green DE . Cardiolipin requirement for electron transfer in complex I and III of the mitochondrial respiratory chain. J Biol Chem 1981; 256: 1874–1880.
Gonzalvez F, Gottlieb E . Cardiolipin: setting the beat of apoptosis. Apoptosis 2007; 12: 877–885.
Houtkooper RH, Vaz FM . Cardiolipin, the heart of mitochondrial metabolism. Cell Mol Life Sci 2008; 65: 2493–2506.
Patil VA, Greenberg ML . Cardiolipin-mediated cellular signaling. Adv Exp Med Biol 2013; 991: 195–213.
Gonzalvez F, Schug ZT, Houtkooper RH, MacKenzie ED, Brooks DG, Wanders RJ et al. Cardiolipin provides an essential activating platform for caspase-8 on mitochondria. J Cell Biol 2008; 183: 681–696.
Schug ZT, Gottlieb E . Cardiolipin acts as a mitochondrial signalling platform to launch apoptosis. Biochim Biophys Acta 2009; 1788: 2022–2031.
Zhang T, Saghatelian A . Emerging roles of lipids in BCL-2 family-regulated apoptosis. Biochim Biophys Acta 2013; 1831: 1542–1554.
Paradies G, Paradies V, De B V, Ruggiero FM, Petrosillo G . Functional role of cardiolipin in mitochondrial bioenergetics. Biochim Biophys Acta 2014; 1837: 408–417.
Barth PG, Scholte HR, Berden JA, Van der Klei-Van Moorsel JM, Luyt-Houwen IE, Van 't Veer-Korthof ET et al. An X-linked mitochondrial disease affecting cardiac muscle, skeletal muscle and neutrophil leucocytes. J Neurol Sci 1983; 62: 327–355.
Houtkooper RH, Rodenburg RJ, Thiels C, van LH, Stet F, Poll-The BT et al. Cardiolipin and monolysocardiolipin analysis in fibroblasts, lymphocytes, and tissues using high-performance liquid chromatography-mass spectrometry as a diagnostic test for Barth syndrome. Anal Biochem 2009; 387: 230–237.
Ren M, Phoon CK, Schlame M . Metabolism and function of mitochondrial cardiolipin. Prog Lipid Res 2014; 55C: 1–16.
Shimabukuro M, Zhou YT, Levi M, Unger RH . Fatty acid-induced beta cell apoptosis: a link between obesity and diabetes. Proc Natl Acad Sci USA 1998; 95: 2498–2502.
Listenberger LL, Han X, Lewis SE, Cases S, Farese Jr RV, Ory DS et al. Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proc Natl Acad Sci USA 2003; 100: 3077–3082.
Mullauer FB, van BL, Daalhuisen JB, Ten Brink MS, Storm G, Medema JP et al. Betulinic acid delivered in liposomes reduces growth of human lung and colon cancers in mice without causing systemic toxicity. Anticancer Drugs 2011; 22: 223–233.
Coleman RA, Lee DP . Enzymes of triacylglycerol synthesis and their regulation. Prog Lipid Res 2004; 43: 134–176.
Kiebish MA, Han X, Cheng H, Chuang JH, Seyfried TN . Cardiolipin and electron transport chain abnormalities in mouse brain tumor mitochondria: lipidomic evidence supporting the Warburg theory of cancer. J Lipid Res 2008; 49: 2545–2556.
Ruggieri S, Roblin R, Black PH . Lipids of whole cells and plasma membrane fractions from Balb/c3T3, SV3T3, and concanavalin A-selected revertant cells. J Lipid Res 1979; 20: 760–771.
Bougnoux P, Chajes V, Lanson M, Hacene K, Body G, Couet C et al. Prognostic significance of tumor phosphatidylcholine stearic acid level in breast carcinoma. Breast Cancer Res Treat 1992; 20: 185–194.
Horie Y, Suzuki A, Kataoka E, Sasaki T, Hamada K, Sasaki J et al. Hepatocyte-specific Pten deficiency results in steatohepatitis and hepatocellular carcinomas. J Clin Invest 2004; 113: 1774–1783.
Pelicano H, Carney D, Huang P . ROS stress in cancer cells and therapeutic implications. Drug Resist Updat 2004; 7: 97–110.
Nogueira V, Hay N . Molecular pathways: reactive oxygen species homeostasis in cancer cells and implications for cancer therapy. Clin Cancer Res 2013; 19: 4309–4314.
Warburg O . On respiratory impairment in cancer cells. Science 1956; 124: 269–270.
Ward PS, Thompson CB . Metabolic reprogramming: a cancer hallmark even warburg did not anticipate. Cancer Cell 2012; 21: 297–308.
Clarke SL, Bowron A, Gonzalez IL, Groves SJ, Newbury-Ecob R, Clayton N et al. Barth syndrome. Orphanet J Rare Dis 2013; 8: 23.
Alavian KN, Beutner G, Lazrove E, Sacchetti S, Park HA, Licznerski P et al. An uncoupling channel within the c-subunit ring of the F1FO ATP synthase is the mitochondrial permeability transition pore. Proc Natl Acad Sci USA 2014; 111: 10580–10585.
Acehan D, Malhotra A, Xu Y, Ren M, Stokes DL, Schlame M . Cardiolipin affects the supramolecular organization of ATP synthase in mitochondria. Biophys J 2011; 100: 2184–2192.
Cory S, Adams JM . The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2002; 2: 647–656.
Tuominen EK, Wallace CJ, Kinnunen PK . Phospholipid-cytochrome c interaction: evidence for the extended lipid anchorage. J Biol Chem 2002; 277: 8822–8826.
Orrenius S, Zhivotovsky B, Nicotera P . Regulation of cell death: the calcium-apoptosis link. Nat Rev Mol Cell Biol 2003; 4: 552–565.
Huttemann M, Pecina P, Rainbolt M, Sanderson TH, Kagan VE, Samavati L et al. The multiple functions of cytochrome c and their regulation in life and death decisions of the mammalian cell: From respiration to apoptosis. Mitochondrion 2011; 11: 369–381.
Kagan VE, Tyurin VA, Jiang J, Tyurina YY, Ritov VB, Amoscato AA et al. Cytochrome c acts as a cardiolipin oxygenase required for release of proapoptotic factors. Nat Chem Biol 2005; 1: 223–232.
Jourdain A, Martinou JC . Mitochondrial outer-membrane permeabilization and remodelling in apoptosis. Int J Biochem Cell Biol 2009; 41: 1884–1889.
Landes T, Martinou JC . Mitochondrial outer membrane permeabilization during apoptosis: the role of mitochondrial fission. Biochim Biophys Acta 2011; 1813: 540–545.
Martinou JC, Youle RJ . Which came first, the cytochrome c release or the mitochondrial fission? Cell Death Differ 2006; 13: 1291–1295.
Pellegrini L, Scorrano L . A cut short to death: Parl and Opa1 in the regulation of mitochondrial morphology and apoptosis. Cell Death Differ 2007; 14: 1275–1284.
Li J, Ding SF, Habib NA, Fermor BF, Wood CB, Gilmour RS . Partial characterization of a cDNA for human stearoyl-CoA desaturase and changes in its mRNA expression in some normal and malignant tissues. Int J Cancer 1994; 57: 348–352.
Lu J, Pei H, Kaeck M, Thompson HJ . Gene expression changes associated with chemically induced rat mammary carcinogenesis. Mol Carcinog 1997; 20: 204–215.
Thai SF, Allen JW, DeAngelo AB, George MH, Fuscoe JC . Detection of early gene expression changes by differential display in the livers of mice exposed to dichloroacetic acid. Carcinogenesis 2001; 22: 1317–1322.
Yahagi N, Shimano H, Hasegawa K, Ohashi K, Matsuzaka T, Najima Y et al. Co-ordinate activation of lipogenic enzymes in hepatocellular carcinoma. Eur J Cancer 2005; 41: 1316–1322.
Morgan-Lappe SE, Tucker LA, Huang X, Zhang Q, Sarthy AV, Zakula D et al. Identification of Ras-related nuclear protein, targeting protein for xenopus kinesin-like protein 2, and stearoyl-CoA desaturase 1 as promising cancer targets from an RNAi-based screen. Cancer Res 2007; 67: 4390–4398.
Scaglia N, Igal RA . Inhibition of Stearoyl-CoA Desaturase 1 expression in human lung adenocarcinoma cells impairs tumorigenesis. Int J Oncol 2008; 33: 839–850.
Fritz V, Benfodda Z, Rodier G, Henriquet C, Iborra F, Avances C et al. Abrogation of de novo lipogenesis by stearoyl-CoA desaturase 1 inhibition interferes with oncogenic signaling and blocks prostate cancer progression in mice. Mol Cancer Ther 2010; 9: 1740–1754.
Roongta UV, Pabalan JG, Wang X, Ryseck RP, Fargnoli J, Henley BJ et al. Cancer cell dependence on unsaturated fatty acids implicates stearoyl-CoA desaturase as a target for cancer therapy. Mol Cancer Res 2011; 9: 1551–1561.
Potze L, Mullauer FB, Colak S, Kessler JH, Medema JP . Betulinic acid-induced mitochondria-dependent cell death is counterbalanced by an autophagic salvage response. Cell Death Dis 2014; 5: e1169.
Waterhouse NJ, Trapani JA . A new quantitative assay for cytochrome c release in apoptotic cells. Cell Death Differ 2003; 10: 853–855.
Valianpour F, Selhorst JJ, van Lint LE, van Gennip AH, Wanders RJ, Kemp S . Analysis of very long-chain fatty acids using electrospray ionization mass spectrometry. Mol Genet Metab 2003; 79: 189–196.
Engelen M, Ofman R, Mooijer PA, Poll-The BT, Wanders RJ, Kemp S . Cholesterol-deprivation increases mono-unsaturated very long-chain fatty acids in skin fibroblasts from patients with X-linked adrenoleukodystrophy. Biochim Biophys Acta 2008; 3: 105–111.
Acknowledgements
We thank Inge Dijkstra, Rob Ofman, Femke Beers-Stet for technical assistance and Louis Vermeulen for his valuable scientific input and helpful discussions. This work was supported by a grant of the Stichting Nationaal Fonds tegen Kanker (SNFK), Amsterdam, The Netherlands (LP).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Potze, L., Di Franco, S., Grandela, C. et al. Betulinic acid induces a novel cell death pathway that depends on cardiolipin modification. Oncogene 35, 427–437 (2016). https://doi.org/10.1038/onc.2015.102
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2015.102
This article is cited by
-
Identification of icaritin derivative IC2 as an SCD-1 inhibitor with anti-breast cancer properties through induction of cell apoptosis
Cancer Cell International (2022)
-
Betulinic acid shows anticancer activity against equine melanoma cells and permeates isolated equine skin in vitro
BMC Veterinary Research (2020)
-
DHRS2 mediates cell growth inhibition induced by Trichothecin in nasopharyngeal carcinoma
Journal of Experimental & Clinical Cancer Research (2019)
-
The natural compound GL22, isolated from Ganoderma mushrooms, suppresses tumor growth by altering lipid metabolism and triggering cell death
Cell Death & Disease (2018)
-
Stearoyl-CoA-desaturase 1 regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ
Oncogene (2017)