1. Use of Human Cancer Cell Lines Mitochondria to Explore the Mechanisms of BH3 Peptides and ABT-737-Induced Mitochondrial Membrane Permeabilization
- Author
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Pierre Rustin, Christine Péchoux, Magali Brabant, Diana Desgué, Etienne Jacotot, Nelly Buron, Myriam Lassalle, Mathieu Porceddu, Annie Borgne-Sanchez, Cindy Racoeur, Theraptosis S.A., Mitologics SAS, Hôpital Robert Debré, Unité de recherche génomique et physiologie de la lactation (GPL), Institut National de la Recherche Agronomique (INRA), AP-HP Hôpital universitaire Robert-Debré [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Diderot - Paris 7 (UPD7), Department of Reproductive Biology, and Imperial College London
- Subjects
Non-Clinical Medicine/Research Methods ,[SDV]Life Sciences [q-bio] ,Mitochondrion ,Mitochondrial apoptosis-induced channel ,Piperazines ,Membrane Potentials ,Nitrophenols ,Mice ,0302 clinical medicine ,BIOLOGIE CELLULAIRE ,Inner mitochondrial membrane ,Mice, Inbred BALB C ,Sulfonamides ,0303 health sciences ,Multidisciplinary ,biology ,Cytochrome c ,Cell Biology/Cellular Death and Stress Responses ,CANCER ,Mitochondria ,3. Good health ,Cell biology ,PROTEINE BCL-2 ,Gene Expression Regulation, Neoplastic ,Cross-Linking Reagents ,Proto-Oncogene Proteins c-bcl-2 ,Biochemistry ,mitochondrie ,protéine ,030220 oncology & carcinogenesis ,Mitochondrial Membranes ,Medicine ,Female ,Biotechnology/Protein Chemistry and Proteomics ,Biochemistry/Drug Discovery ,biological phenomena, cell phenomena, and immunity ,Bacterial outer membrane ,Research Article ,Science ,Oncology/Oncology Agents ,Permeability ,03 medical and health sciences ,Cell Line, Tumor ,Proto-Oncogene Proteins ,Animals ,Humans ,[INFO]Computer Science [cs] ,030304 developmental biology ,apoptose ,Biphenyl Compounds ,Peptide Fragments ,CYTOCHROME-C RELEASE ,BCL-2 FAMILY PROTEINS ,MIMETIC ABT-737 ,OUTER-MEMBRANE ,CASPASE ACTIVATION ,BAX ,APOPTOSIS ,DEATH ,INHIBITORS ,DOMAINS ,membrane cellulaire ,Apoptosis ,Cancer cell ,biology.protein ,Apoptosome ,humain - Abstract
International audience; Current limitations of chemotherapy include toxicity on healthy tissues and multidrug resistance of malignant cells. A number of recent anti-cancer strategies aim at targeting the mitochondrial apoptotic machinery to induce tumor cell death. In this study, we set up protocols to purify functional mitochondria from various human cell lines to analyze the effect of peptidic and xenobiotic compounds described to harbour either Bcl-2 inhibition properties or toxic effects related to mitochondria. Mitochondrial inner and outer membrane permeabilization were systematically investigated in cancer cell mitochondria versus non-cancerous mitochondria. The truncated (t-) Bid protein, synthetic BH3 peptides from Bim and Bak, and the small molecule ABT-737 induced a tumor-specific and OMP-restricted mitochondrio-toxicity, while compounds like HA-14.1, YC-137, Chelerythrine, Gossypol, TW-37 or EM20-25 did not. We found that ABT-737 can induce the Bax-dependent release of apoptotic proteins (cytochrome c, Smac/Diablo and Omi/HtrA2 but not AIF) from various but not all cancer cell mitochondria. Furthermore, ABT-737 addition to isolated cancer cell mitochondria induced oligomerization of Bax and/or Bak monomers already inserted in the mitochondrial membrane. Finally immunoprecipatations indicated that ABT-737 induces Bax, Bak and Bim desequestration from Bcl-2 and Bcl-xL but not from Mcl-1L. This study investigates for the first time the mechanism of action of ABT-737 as a single agent on isolated cancer cell mitochondria. Hence, this method based on MOMP (mitochondrial outer membrane permeabilization) is an interesting screening tool, tailored for identifying Bcl-2 antagonists with selective toxicity profile against cancer cell mitochondria but devoid of toxicity against healthy mitochondria.
- Published
- 2010