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KMID : 0613820230330010015
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Journal of Life Science
2023 Volume.33 No. 1 p.15 ~ p.24
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Apoptotic Effect of Proso Millet Grains on Human Breast Cancer MDA-MB-231 Cells Is Exerted by Activation of BAK and BAX, and Mitochondrial Damage- mediated Caspase Cascade Activation
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Jun Do-Youn
Han Cho-Rong
Kim Young-Ho
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Abstract
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To examine the antitumor effect of proso millet grains, whether proso millet grains exert apoptotic activity against human cancer cells was investigated. When the cytotoxicity of 80% ethanol (EtOH) extract of proso millet grains was tested against various cancer cells using MTT assay, more potent cytotoxicity was observed against human breast cancer MDA-MB-231 cells than against other cancer cells. When the EtOH extract was evaporated to dryness, dissolved in water, and then further fractio- nated by sequential extraction using four organic solvents (n-hexane, methylene chloride, ethyl acetate, and n-butanol), the BuOH fraction exhibited the highest cytotoxicity against MDA-MB-231 cells.
Along with the cytotoxicity, TUNEL-positive apoptotic nucleosomal DNA fragmentation and several apoptotic responses including BAK/BAX activation, mitochondria membrane potential (Δψm) loss, mitochondrial cytochrome c release into the cytosol, activation of caspase-8/-9/-3, and degradation of poly (ADP-ribose) polymerase (PARP) were detected. However, human normal mammary epithelial MCF-10A cells exhibited a significantly lesser extent of sensitivity compared to malignant MDA-MB231 cells. Irrespective of Fas-associated death domain (FADD)-deficiency or caspase-8-deficiency,
human T acute lymphoblastic leukemia Jurkat cells displayed similar sensitivities to the cytotoxicity of BuOH fraction, excluding an involvement of extrinsic apoptotic mechanism in the apoptosis induction. These results demonstrate that the cytotoxicity of BuOH fraction from proso millet grains against human breast cancer MDA-MB-231 cells is attributable to intrinsic apoptotic cell death resulting from BAK/BAX activation, and subsequent mediation of mitochondrial damage-dependent activation of caspase cascade.
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KEYWORD
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Activation of BAK and BAX, breast cancer cells, butanol fraction, mitochondrial apoptosis, proso millet grains
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