title: Lonidamine triggers apoptosis via a direct, Bcl-2-inhibited effect on the mitochondrial permeability transition pore.
abstract: The molecular mode of action of lonidamine, a therapeutic agent employed in cancer chemotherapy, has been elusive. Here we provide evidence that lonidamine (LND) acts on mitochondria to induce apoptosis. LND provokes a disruption of the mitochondrial transmembrane potential which precedes signs of nuclear apoptosis and cytolysis. The mitochondrial and cytocidal effects of LND are not prevented by inhibitors of caspases or of mRNA or protein synthesis. However, they are prevented by transfection-enforced overexpression of Bcl-2, an oncoprotein which inhibits apoptosis by stabilizing the mitochondrial membrane barrier function. Accordingly, the cell death-inducing effect of LND is amplified by simultaneous addition of PK11195, an isoquinoline ligand of the peripheral benzodiazepine receptor which antagonizes the cytoprotective effect of Bcl-2. When added to isolated nuclei, LND fails to provoke DNA degradation unless mitochondria are added simultaneously. In isolated mitochondria, LND causes the dissipation of the mitochondrial inner transmembrane potential and the release of apoptogenic factors capable of inducing nuclear apoptosis in vitro. Thus the mitochondrion is the subcellular target of LND. All effects of LND on isolated mitochondria are counteracted by cyclosporin A, an inhibitor of the mitochondrial PT pore. We therefore tested the effect of LND on the purified PT pore reconstituted into liposomes. LND permeabilizes liposomal membranes containing the PT pore. This effect is prevented by addition of recombinant Bcl-2 protein but not by a mutant Bcl-2 protein that has lost its apoptosis-inhibitory function. Altogether these data indicate that LND represents a novel type of anti-cancer agent which induces apoptosis via a direct effect on the mitochondrial PT pore.
authors.affiliation: Centre National de la Recherche Scientifique, Unité Propre de Recherche 420, Villejuif, France.
authors.forename: L
authors.initials: L
authors.lastname: Ravagnan
authors.forename: I
authors.initials: I
authors.lastname: Marzo
authors.forename: P
authors.initials: P
authors.lastname: Costantini
authors.forename: S A
authors.initials: SA
authors.lastname: Susin
authors.forename: N
authors.initials: N
authors.lastname: Zamzami
authors.forename: P X
authors.initials: PX
authors.lastname: Petit
authors.forename: F
authors.initials: F
authors.lastname: Hirsch
authors.forename: M
authors.initials: M
authors.lastname: Goulbern
authors.forename: M F
authors.initials: MF
authors.lastname: Poupon
authors.forename: L
authors.initials: L
authors.lastname: Miccoli
authors.forename: Z
authors.initials: Z
authors.lastname: Xie
authors.forename: J C
authors.initials: JC
authors.lastname: Reed
authors.forename: G
authors.initials: G
authors.lastname: Kroemer
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D002474:Cell-Free System; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D006801:Humans; D007191:Indazoles / Q000494:pharmacology*; D007473:Ion Channels / Q000187:drug effects* / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D010539:Permeability; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000378:metabolism; D020298:U937 Cells
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D000970:Antineoplastic Agents; D015853:Cysteine Proteinase Inhibitors; D007191:Indazoles; D007473:Ion Channels; D007546:Isoquinolines; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C016371:lonidamine; C037850:PK 11195