title: Mitochondria are the target organelle of differentiation-inducing factor-3, an anti-tumor agent isolated from Dictyostelium discoideum [corrected].
abstract: Differentiation-inducing factor-3 (DIF-3), found in the cellular slime mold Dictyostelium discoideum, and its derivatives such as butoxy-DIF-3 (Bu-DIF-3) are potent anti-tumor agents. However, the precise mechanisms underlying the actions of DIF-3 remain to be elucidated. In this study, we synthesized a green fluorescent derivative of DIF-3, BODIPY-DIF-3, and a control fluorescent compound, Bu-BODIPY (butyl-BODIPY), and investigated how DIF-like molecules behave in human cervical cancer HeLa cells by using both fluorescence and electron microscopy. BODIPY-DIF-3 at 5-20 µ M suppressed cell growth in a dose-dependent manner, whereas Bu-BODIPY had minimal effect on cell growth. When cells were incubated with BODIPY-DIF-3 at 20 µM, it penetrated cell membranes within 0.5 h and localized mainly in mitochondria, while Bu-BODIPY did not stain the cells. Exposure of cells for 1-3 days to DIF-3, Bu-DIF-3, BODIPY-DIF-3, or CCCP (a mitochondrial uncoupler) induced substantial mitochondrial swelling, suppressing cell growth. When added to isolated mitochondria, DIF-3, Bu-DIF-3, and BOIDPY-DIF-3, like CCCP, dose-dependently promoted the rate of oxygen consumption, but Bu-BODIPY did not. Our results suggest that these bioactive DIF-like molecules suppress cell growth, at least in part, by disturbing mitochondrial activity. This is the first report showing the cellular localization and behavior of DIF-like molecules in mammalian tumor cells.
authors: Department of Molecular and Cellular Biology, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Japan.
authors: Yuzuru
authors: Y
authors: Kubohara
authors: Haruhisa
authors: H
authors: Kikuchi
authors: Yusuke
authors: Y
authors: Matsuo
authors: Yoshiteru
authors: Y
authors: Oshima
authors: Yoshimi
authors: Y
authors: Homma
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D001896:Boron Compounds / Q000737:chemistry / Q000378:metabolism; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002463:Cell Membrane Permeability; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D056741:Cyclin D / Q000378:metabolism; D004023:Dictyostelium / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006588:Hexanones / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D010101:Oxygen Consumption; D061209:Proton Ionophores / Q000494:pharmacology
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000970:Antineoplastic Agents; D001896:Boron Compounds; D056741:Cyclin D; D005456:Fluorescent Dyes; D006588:Hexanones; D061209:Proton Ionophores; C055364:1-(3-chloro-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone
references: Cancer Res. 1989 Aug 15;49(16):4477-82
references: 2743336
references: Br J Cancer. 1984 Feb;49(2):225-33
references: 6199034
references: Biochem Biophys Res Commun. 1995 Mar 28;208(3):1036-9
references: 7702602
references: Int J Cell Cloning. 1987 May;5(3):191-201
references: 2439615
references: PLoS One. 2009 Aug 17;4(8):e6658
references: 19684855
references: J Cell Physiol. 1985 Sep;124(3):554-6
references: 4044664
references: J Org Chem. 2005 Oct 28;70(22):8854-8
references: 16238318
references: Am J Physiol Cell Physiol. 2010 Jan;298(1):C107-13
references: 19889959
references: Toxicol Lett. 2012 Jun 20;211(3):312-8
references: 22561169
references: Biochimie. 2002 Feb-Mar;84(2-3):103-4
references: 12022940
references: Science. 1998 Aug 28;281(5381):1309-12
references: 9721092
references: Biochem Biophys Res Commun. 1997 Jul 18;236(2):418-22
references: 9240452
references: Eur J Pharmacol. 1999 Sep 17;381(1):57-62
references: 10528134
references: Circ Res. 2000 Jan 7-21;86(1):68-75
references: 10625307
references: Br J Cancer. 1992 Aug;66(2):311-7
references: 1503904
references: Biochem J. 1988 Feb 1;249(3):903-6
references: 3355503
references: Bioorg Med Chem. 2004 Jun 15;12(12):3203-14
references: 15158788
references: Oncogene. 2003 Jan 30;22(4):548-54
references: 12555068
references: Eur J Pharmacol. 2004 Feb 6;485(1-3):21-9
references: 14757120
references: Biochem Pharmacol. 2005 Sep 1;70(5):676-85
references: 16023080
references: J Biomol Screen. 2011 Sep;16(8):818-24
references: 21788393
references: Br J Pharmacol. 2006 Feb;147(3):239-48
references: 16331284
references: Int J Cancer. 1998 May 4;76(3):418-22
references: 9579581
references: Nature. 1987 Aug 27-Sep 2;328(6133):811-4
references: 3627228
references: Science. 2009 Feb 6;323(5915):793-7
references: 19131594
references: J Transl Med. 2009 Feb 05;7:13
references: 19196452
references: Mol Cell. 2010 Jan 15;37(1):79-89
references: 20129057
references: Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5660-3
references: 7777565
references: J Biol Chem. 2003 Mar 14;278(11):9663-70
references: 12522140
references: J Biol Chem. 2006 Dec 15;281(50):38489-97
references: 17046823
references: Life Sci. 2006 Dec 14;80(2):160-5
references: 17027864
references: FEBS Lett. 1995 Feb 13;359(2-3):119-22
references: 7867781
references: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12801-6
references: 12228733
references: Science. 2004 Jul 30;305(5684):626-9
references: 15286356
references: Semin Cell Dev Biol. 1999 Dec;10(6):577-85
references: 10706822
references: Cancer Res. 2004 Apr 1;64(7):2568-71
references: 15059913
references: Development. 1989;107 Suppl:81-90
references: 2699860
references: J Pharmacol Sci. 2010;112(3):320-6
references: 20173310