Abstract 1021: Use of functional tumor cell line mitochondria to explore the mechanisms of BH3 peptides and ABT-737-induced mitochondrial membrane permeabilization

Abstract

Abstract Current limitations of chemotherapy include toxicity on healthy tissues and multidrug resistance on malignant cells. A number of recent anti-cancer strategies are targeting the mitochondrial apoptotic machinery to induce tumor cell death. In this study, we set up protocols to isolate mitochondria from various origins with high purity, quality and functionality. We analyzed the effect of putative Bcl-2 inhibitors on both healthy and tumor isolated mitochondria. We measured mitochondrial membrane permeabilization (swelling), mitochondrial transmembrane potential (ΔΨm) and mitochondrial outer membrane permeabilization (MOMP) in both isolated healthy and tumoral cell mitochondria. Like t-Bid and some BH3 peptides, the molecule ABT-737 was found to present a tumor-specific mitochondrio-toxicity, while compounds like HA-14.1, YC-137, Chelerythrine, Gossypol, TW-37 or EM20-25 did not. We thus demonstrated that ABT-737 can induce a relatively large and targeted MOMP triggering the release of apoptotic proteins from tumoral cell mitochondria (Cytochrome c, Smac/Diablo and Omi/HtrA2 but not AIF) however without any mitochondrial swelling. Furthermore, ABT-737 addition to isolated tumoral cell mitochondria induced specific Bax/Bak channel formation by oligomerizing monomeric Bax and/or Bak already inserted in the mitochondrial membrane. We also provided evidence that tumoral cell mitochondria can be either resistant or sensitive to ABT-737-induced MOMP, depending on their origin and interactions between pro- and anti-apoptotic proteins. Finally, this method based on MOMP is an interesting screening tool, tailored for identifying Bcl-2 antagonists with selective toxicity profile against tumoral cell mitochondria but devoid of toxicity against healthy mitochondria. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1021.</jats:p