11
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Securinine from Phyllanthus glaucus Induces Cell Cycle Arrest and Apoptosis in Human Cervical Cancer HeLa Cells

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Background

          The Securinega-type alkaloids occur in plants belonging to Euphorbiaceae family. One of the most widely distributed alkaloid of this group is securinine, which was identified next to allosecurinine in Phyllanthus glaucus (leafflower). Recently, some Securinega-type alkaloids have paid attention to its antiproliferative potency towards different cancer cells. However, the cytotoxic properties of allosecurinine have not yet been evaluated.

          Methods

          The cytotoxicity of the extract, alkaloid fraction obtained from P. glaucus, isolated securinine and allosecurinine against HeLa cells was evaluated by real-time xCELLigence system and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected by annexin V and 7-amino-actinomycin (7-AAD) staining and confirmed with fluorescent Hoechst 33342 dye. The assessment of mitochondrial membrane potential (MMP), reactive oxygen species (ROS) generation, the level of extracellular signal-regulated protein kinases 1 and 2 (ERK1/2), caspase-3/7 activity and cell cycle analysis were measured by flow cytometry. The enzymatic activity of caspase-9 was assessed by a luminometric assay. The expression of apoptosis associated genes was analyzed by real-time PCR.

          Results

          The experimental data revealed that securinine and the alkaloid fraction were significantly potent on HeLa cells growth inhibition with IC 50 values of 7.02 ± 0.52 μg/ml (32.3 μM) and 25.46 ± 1.79 μg/ml, respectively. The activity of allosecurinine and Phyllanthus extract were much lower. Furthermore, our study showed that the most active securinine induced apoptosis in a dose-dependent manner in the tested cells, increased the percentage of ROS positive cells and depolarized cells as well as stimulated the activity of ERK1/2, caspase-9 and -3/7. Securinine also induced cell cycle arrest in S phase. Real-time PCR analysis showed high expression of TNFRSF genes in the cells stimulated with securinine.

          Conclusions

          Securinine induces apoptosis and activates cell cycle checkpoints in HeLa cells which is associated with oxidative stress. The results indicate that the mitochondrial pathway is involved in the programmed cell death.

          Related collections

          Most cited references20

          • Record: found
          • Abstract: found
          • Article: not found

          Cell cycle checkpoint signaling: cell cycle arrest versus apoptosis.

          Although toxicants may initiate cell damage or stress, the cellular proteins that are involved in control of cell cycle and apoptosis are the final arbiters of cell fate. The biochemical pathways that restrain cell cycle transition and/or induce cell death after stress are known as cell cycle checkpoints. These checkpoints maintain the fidelity of DNA replication, repair, and division. Herein, select cell cycle checkpoint signaling pathways will be discussed and how different components of these pathways are regulated by exogenous and endogenous agents, with focus on the p53 tumor suppressor signaling. The p53 protein is known to play a key role in growth arrest and apoptosis after cell stress, primarily through its ability to regulate the transcription of select downstream target genes in the cell. Further elucidation of the signaling pathways that control growth arrest and apoptosis will continue to provide insights to the complex cellular responses to environmental toxicants.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Mitochondria and cell death. Mechanistic aspects and methodological issues.

            Mitochondria are involved in cell death for reasons that go beyond ATP supply. A recent advance has been the discovery that mitochondria contain and release proteins that are involved in the apoptotic cascade, like cytochrome c and apoptosis inducing factor. The involvement of mitochondria in cell death, and its being cause or consequence, remain issues that are extremely complex to address in situ. The response of mitochondria may critically depend on the type of stimulus, on its intensity, and on the specific mitochondrial function that has been primarily perturbed. On the other hand, the outcome also depends on the integration of mitochondrial responses that cannot be dissected easily. Here, we try to identify the mechanistic aspects of mitochondrial involvement in cell death as can be derived from our current understanding of mitochondrial physiology, with special emphasis on the permeability transition and its consequences (like onset of swelling, cytochrome c release and respiratory inhibition); and to critically evaluate methods that are widely used to monitor mitochondrial function in situ.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              A death-promoting role for extracellular signal-regulated kinase.

              Extracellular signal-regulated protein kinases 1 and 2 (ERK1/2), which are members of the mitogen-activated protein kinase superfamily, have been well characterized and are known to be involved in cell survival; however, recent evidence suggests that the activation of ERK1/2 also contributes to cell death in some cell types and organs under certain conditions. For example, ERK1/2 is activated in neuronal and renal epithelial cells upon exposure to oxidative stress and toxicants and deprivation of growth factors, and inhibition of the ERK pathway blocks apoptosis. ERK activation also occurs in animal models of ischemia- and trauma-induced brain injury and cisplatin-induced renal injury, and inactivation of ERK reduces the extent of tissue damage. In some studies, ERK has been implicated in apoptotic events upstream of mitochondrial cytochrome c release, whereas other studies have suggested the converse that ERK acts downstream of mitochondrial events and upstream of caspase-3 activation. ERK also can contribute to cell death through the suppression of the antiapoptotic signaling molecule Akt. Here we summarize the evidence and mechanism of ERK-induced apoptosis in both cell culture and in animal models.
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                28 October 2016
                2016
                : 11
                : 10
                : e0165372
                Affiliations
                [1 ]Department of Biology and Pharmaceutical Botany, Medical University of Gdańsk, Gdańsk, Poland
                [2 ]Department of Pharmacognosy with Medicinal Plant Garden, Medical University of Gdańsk, Gdańsk, Poland
                University of PECS Medical School, HUNGARY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: JRO MKB BSS JSH.

                • Data curation: JSH BSS.

                • Formal analysis: JSH BSS.

                • Funding acquisition: BSS JRO MKB.

                • Investigation: JSH BSS.

                • Methodology: JSH BSS MKB JRO.

                • Resources: JSH BSS JRO MKB.

                • Software: JSH BSS.

                • Supervision: JRO MKB.

                • Validation: JSH BSS.

                • Visualization: JSH.

                • Writing – original draft: JSH BSS.

                • Writing – review & editing: JRO MKB.

                Article
                PONE-D-16-23324
                10.1371/journal.pone.0165372
                5085043
                27792748
                53e6ff38-7b6a-4e14-aa10-c6d5ccea9fce
                © 2016 Stefanowicz-Hajduk et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 10 June 2016
                : 11 October 2016
                Page count
                Figures: 11, Tables: 1, Pages: 19
                Funding
                Funded by: National Science Centre
                Award ID: DEC-2012/05/N/NZ/00957
                Award Recipient :
                Funded by: the Ministry of Science and Higher Education of the Republic of Poland
                Award ID: the Leading National Research Centre (KNOW) program for the years 2012–2017
                Award Recipient :
                The project was financed by a (NCN) National Science Centre grant (DEC-2012/05/N/NZ/00957). This project was supported by the Ministry of Science and Higher Education of the Republic of Poland, from the quality—promoting subsidy, under the Leading National Research Centre (KNOW) program for the years 2012–2017.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Research and analysis methods
                Biological cultures
                Cell lines
                HeLa cells
                Research and analysis methods
                Biological cultures
                Cell cultures
                Cultured tumor cells
                HeLa cells
                Physical Sciences
                Chemistry
                Chemical Compounds
                Alkaloids
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Cycle and Cell Division
                Biology and Life Sciences
                Biochemistry
                Bioenergetics
                Energy-Producing Organelles
                Mitochondria
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Energy-Producing Organelles
                Mitochondria
                Research and analysis methods
                Bioassays and physiological analysis
                Biochemical analysis
                Colorimetric assays
                MTT assay
                Research and analysis methods
                Bioassays and physiological analysis
                Biochemical analysis
                Enzyme assays
                MTT assay
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Cycle and Cell Division
                Synthesis Phase
                Biology and Life Sciences
                Cell Biology
                Oxidative Stress
                Custom metadata
                All relevant data are within the paper and its Supporting Information Files.

                Uncategorized
                Uncategorized

                Comments

                Comment on this article