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      Methyl Angolensate from Callus of Indian Redwood Induces Cytotoxicity in Human Breast Cancer Cells

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          Abstract

          AIM:

          Natural products discovered from medicinal plants have played an important role in the treatment of cancer. Methyl angolensate (MA), a tetranortriterpenoid obtained from the root callus of Indian Redwood tree, Soymida febrifuga Roxb. (A.Juss) was tested for its anticancer properties on breast cancer cells.

          METHODS:

          Cell viability was tested using trypan blue, MTT and LDH assays. Tritiated thymidine assay and flowcytometry were used to study effect of MA on cell proliferation. The activation of apoptosis was checked by annexin V and JC-1 staining followed by FACS analysis. Immunoblotting analysis was used for studying expression of apoptotic and DNA double strand break repair proteins.

          RESULTS:

          We find that MA inhibited the growth of breast cancer cell line, T47D in a time- and dose-dependent manner. MA treatment led to the inhibition of cell proliferation as detected by tritiated thymidine assay and flowcytometry. Further, MA treated cells exhibited typical apoptotic morphological changes and led to the accumulation of subG1 peak in cell cycle distribution. The induction of apoptosis was further confirmed both by annexin V staining and JC1 staining. We also find that MA activates MAP kinase pathway to induce apoptosis. Besides, we find a time dependent activation followed by degradation of DNA double-strand break repair proteins upon treatment with MA.

          CONCLUSION:

          These results suggest that MA induces cytotoxicity in breast cancer cells. Further, the altered expression of DSB repair proteins in MA treated cells may control the induction of apoptosis in these cancer cells.

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          Most cited references41

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          Mitochondria and apoptosis.

          D Green, J Reed (1998)
          A variety of key events in apoptosis focus on mitochondria, including the release of caspase activators (such as cytochrome c), changes in electron transport, loss of mitochondrial transmembrane potential, altered cellular oxidation-reduction, and participation of pro- and antiapoptotic Bcl-2 family proteins. The different signals that converge on mitochondria to trigger or inhibit these events and their downstream effects delineate several major pathways in physiological cell death.
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            Mitogen-activated protein kinases: specific messages from ubiquitous messengers.

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              Apoptosis-inducing factor (AIF): key to the conserved caspase-independent pathways of cell death?

              Numerous pro-apoptotic signal transducing molecules act on mitochondria and provoke the permeabilization of the outer mitochondrial membrane, thereby triggering the release of potentially toxic mitochondrial proteins. One of these proteins, apoptosis-inducing factor (AIF), is a phylogenetically old flavoprotein which, in healthy cells, is confined to the mitochondrial intermembrane space. Upon lethal signaling, AIF translocates, via the cytosol, to the nucleus where it binds to DNA and provokes caspase-independent chromatin condensation. The crystal structures of both human and mouse AIF have been determined, and the fine mechanisms accounting for its oxidoreductase activity and its electrostatic interaction with double-stranded DNA have been elucidated. Importantly, the apoptogenic and oxidoreductase functions of AIF can be dissociated. Thus, mutations that abolish the AIF-DNA interaction suppress AIF-induced chromatin condensation, yet have no effect on the NADH oxidase activity. Recent studies suggest AIF to be a major factor determining caspase-independent neuronal death, emphasizing the central role of mitochondria in the control of physiological and pathological cell demise.
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                Author and article information

                Journal
                Int J Biomed Sci
                Int J Biomed Sci
                IJBS
                International Journal of Biomedical Science : IJBS
                Master Publishing Group
                1550-9702
                1555-2810
                September 2010
                : 6
                : 3
                : 182-194
                Affiliations
                Department of Biochemistry, Indian Institute of Science, Bangalore, India
                Author notes
                Corresponding author: Sathees C. Raghavan, Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012, India. Tel: 080 2293 2674; Fax: 080 2360 0814; E-mail: sathees@ 123456biochem.iisc.ernet.in .
                Article
                IJBS-6-182
                3615260
                23675192
                69a008a9-a8d2-4a07-8305-27084d47c7d3
                © Kishore K. Chiruvella et al. Licensee Master Publishing Group

                This is an open-access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.5/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 14 May 2010
                : 7 June 2010
                Categories
                Article

                double-strand breaks,intrinsic pathway of apoptosis,cancer therapeutics,alternative medicine,nonhomologous dna end-joining

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