Mitochondrial Delivery of Bongkrekic Acid Using a MITO-Porter Prevents the Induction of Apoptosis in Human HeLa Cells
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INTRODUCTION
Apoptosis is a process of programmed cell death and is a major mechanism involved in the precise regulation of cell numbers and functions as a defense mechanism to remove unwanted and potentially dangerous cells. Mitochondria play a central role in the regulation of apoptosis1., 2. and mitotchondrial dysfunction causes a variety of human disorders.3 Therefore, mitochondria represent an important target organelle for drug therapy,4., 5. and thus therapy promises to be useful and productive for
Materials
1,2-Dioleoyl-sn-glycero-3-phosphatidyl ethanolamine (DOPE) and 7-nitrobenz-2-oxa-1, 3-diazole labeled DOPE (NBD-DOPE) were purchased from Avanti Polar lipids (Alabaster, Alabama). Sphingomyelin (SM)was purchased from Sigma (St. Louis, MO). Stearyl octaarginine (STR-R8)22 was obtained from Kurabo Industries Ltd. (Osaka, Japan). HeLa human cervix carcinoma cells were obtained from RIKEN Cell Bank (Tsukuba, Japan). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and
Construction of BKA-MITO-Porter
We first examined the effect of BKA on the MITO- Porter preparation, where mitochondria-fusogenic lipid composition [DOPE/SM/STR-R8 = 9:2:1 (molar ratio)]17., 19. was used. The BKA–MITO-Porter was prepared using lipid films containing different amounts of BKA (0–40 mol % of total lipid) (Fig 2a). The diameters of the MITO-Porter with different contents of BKA were comparable up to a content ratio of 20 mol% BKA (about 100 nm), while aggregation occurred at higher content of BKA (Fig. 2b, Table S1).
DISCUSSION
Previous studies reported that BKA has a high affinity for the ATP–ADP translocator protein in mitochondria, resulting in the inhibition of mitochondrial oxidative phosphorylation, that is, BKA is clearly a toxic chemical.7., 8. On the other hand, BKA has also been found to be an inhibitor of apoptosis.9., 10., 11. Chen et al.28 recently showed that BKA blocked the loss of mitochondrial membrane potential when apoptosis was induced in cells that had been pretreated with BKA. In this case, it
CONCLUSIONS
In this study, we reported on an approach for the mitochondrial delivery of BKA, an antiapoptosis agent, using a MITO-Porter. We first developed a BKA–MITO-Porter and showed that it is taken up by cells and delivered to mitochondria, similar to that of the conventional MITO-Porter. In addition, the mitochondrial delivery of BKA using the MITO-Porter resulted in an antiapoptosis effect in HeLa cells. These results provide support for the assumption that the mitochondrial delivery of BKA can be
ACKNOWLEDGMENTS
This work was supported, in part by the Advanced research for medical products Mining Programme of the National Institute of Biomedical Innovation (NIBIO), a grant-in-aid for Young Scientists (A) and a grant-in-aid for Scientific Research (S) from the Ministry of Education, Culture, Sports, Science and Technology of Japanese Government (MEXT). We also thank Dr. Milton Feather for his helpful advice in writing the manuscript.
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2018, Journal of Controlled ReleaseCitation Excerpt :All animal protocols were approved by the institutional animal care and research advisory committee of the Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan (date: 28 March 2016, registration no. 16-0015). We prepared the RES-MITO-Porter by the lipid film hydration method as previously reported [18]. The RES-MITO-Porter contains resveratrol in the lipid-layer of the carrier.
Mitochondrial Delivery of Doxorubicin Using MITO-Porter Kills Drug-Resistant Renal Cancer Cells via Mitochondrial Toxicity
2017, Journal of Pharmaceutical SciencesCitation Excerpt :The MITO-Porter is an LP-based mitochondrial delivery system that functions via membrane fusion.14,16 To date, we have shown that the MITO-Porter can be used to deliver a variety of therapeutic cargoes, including an antiapoptosis chemical and an antioxidant chemical, to mitochondria of human cells.17-19 As a result, the mitochondrial delivery of therapeutic cargoes have the potential for functioning as a mitochondrial therapeutic strategy in in vitro experiments.
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2016, Journal of Controlled ReleaseCitation Excerpt :To achieve these goals, our research then focused on the development of a mitochondrial medicine based on an established MITO-Porter system. We first attempted to validate the therapeutic strategy using model disease cells in in vitro experiments that involved the mitochondrial delivery of superoxide dismutase [106], an anti-oxidant enzyme and bongkrekic acid [107], an anti-apoptosis chemical. The result indicated that the MITO-Porter represents a potentially useful carrier for use as a mitochondrial medicine, based on the results of in vitro experiments [106,107].
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