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Oligonucleosomal DNA Fragmentation in MCF-7 Cells Undergoing Palmitate-Induced Apoptosis

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Abstract

Oligonucleosomal fragmentation of nuclear DNA is the late-stage apoptosis hallmark. In apoptotic mammalian cells the fragmentation is catalyzed by DFF40/CAD DNase primarily activated by caspase 3 through the site-specific proteolytic cleavage of DFF45/ICAD. A deletion in the casp3 gene of human breast adenocarcinoma MCF-7 results in lack of procaspase 3 in these cells. The absence of caspase 3 in MCF-7 leads to disability to activate oligonucleosomal DNA fragmentation in TNF-α induced cell death. In this study, sodium palmitate was used as an apoptotic stimulus for MCF-7. It has been shown that palmitate but not TNF-α induces both apoptotic changes in nuclei and oligonucleosomal DNA fragmentation in casp3-mutated MCF-7. Activation and accumulation of 40-50 kD DFF40-like DNases in nuclei of palmitate-treated apoptotic MCF-7 were detected by SDS-DNA-PAGE assay. Microsomal fraction of apoptotic MCF-7 does not contain any detectable DNases, but activates 40-50 kD nucleases when incubated with human placental chromatin. Furthermore, microsomes of apoptotic MCF-7 induce oligonucleosomal fragmentation of chromatin in a cell-free system. Both the activation of DNases and chromatin fragmentation are suppressed in the presence of the caspase 3/7 inhibitor Ac-DEVD-CHO. Microsome-associated caspase 7 is suggested to play an essential role in the induction of oligonucleosomal DNA fragmentation in casp3-deficient MCF-7 cells.

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Authors and Affiliations

  1. Siberian Division of the Russian Academy of Sciences, pr. Lavrentieva 8, Novosibirsk Institute of Bioorganic Chemistry, Novosibirsk, 630090, Russia

    D. V. Semenov, P. A. Aronov, E. V. Kuligina, M. O. Potapenko & V. A. Richter

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  1. D. V. Semenov
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  2. P. A. Aronov
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  3. E. V. Kuligina
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  4. M. O. Potapenko
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  5. V. A. Richter
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Semenov, D.V., Aronov, P.A., Kuligina, E.V. et al. Oligonucleosomal DNA Fragmentation in MCF-7 Cells Undergoing Palmitate-Induced Apoptosis. Biochemistry (Moscow) 68, 1335–1341 (2003). https://doi.org/10.1023/B:BIRY.0000011655.58235.44

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  • DOI: https://doi.org/10.1023/B:BIRY.0000011655.58235.44

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