Phcog.Net logo
Submit

BROWSE ALL JOURNALS

    SEE ALL 6 JOURNALS
    Pharmacognosy MagazineNOW IN
    Article

    Clinacanthus nutans Induced Reactive Oxygen Species‑dependent Apoptosis and Autophagy in HCT116 Human Colorectal Cancer Cells

    Kar Suen Wang1★, Chim Kei Chan1, Ahmad Fadhlurrahman Ahmad Hidayat1, Yau Hsiung Wong2, Habsah Abdul Kadir1★ Corresponding author

    1. 1Institute of Biological Science, Faculty of Science, Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia.
    2. 2Department of Mathematics and Computer Science, School of Liberal Art and Sciences, Taylor’s University Lakeside Campus, Subang Jaya, Selangor Darul Ehsan, Malaysia.

    CORRESPONDENCE

    Kar Suen Wang

    Institute of Biological Science, Faculty of Science, Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia.

    Received: 08-07-2017; Revised: 16-08-2017.

    Volume 15, Issue 60 · pp. 87–97 · PUBLISHED 23 January 2019 · DOI: 10.4103/pm.pm_299_17

    View on Pharmacogn. Mag. original site ↗

    ABSTRACT

    Objective: Our study aims to investigate the apoptosis‑ and autophagy‑inducing effects of C. nutans in HCT116 human colorectal cancer cells. Materials and Methods: Cytotoxicity of ethanol extract, hexane, ethyl acetate, and aqueous fractions of C. nutans against various cancer cell lines was determined via MTT assay. Apoptosis assays including annexin V, mito‑ID, and Hoechst 33342/propidium iodide staining were carried out. The level of intracellular reactive oxidative species was determined using flow cytometry. Western blot analysis was carried out to assess the protein expression in C. nutans ethyl acetate fraction (CNEAF)‑treated HCT116 cells. Results: CNEAF was found to exert the strongest cytotoxic effect against HCT116 cells (IC50 = 48.81 ± 1.44 µg/mL). CNEAF‑induced apoptosis was evidenced by nuclear morphological alterations, phosphatidylserine externalization, dissipation of mitochondrial membrane potential, and elevation of intracellular reactive oxygen species (ROS) level. Dissipation of mitochondrial membrane potential was attributed to the upregulation of Bax and Bak accompanied by downregulation of Bcl‑2 and Bcl‑xL, leading to caspase‑3, ‑9, ‑8, and ‑10 activation. Interestingly, an upregulation of death receptor 5 was detected, suggesting involvement of intrinsic and extrinsic pathways. In addition, the occurrence of autophagy by CNEAF was supported by LC‑3 accumulation and p62 degradation. The reduction of intracellular ROS level by N‑acetylcysteine showed that the apoptosis and autophagy induced by CNEAF is ROS dependent. Conclusions: CNEAF induced ROS‑dependent apoptosis and autophagy on HCT116 cells.

    KEYWORDS

    Open in new tab

    REFERENCES

    As published

    Showing references and in-text citations exactly as published.

      Cite this article

      SELECT FORMAT

      Wang, K. S., Chan, C. K., Hidayat, A. F. A., Wong, Y. H., & Kadir, H. A. (2019). Clinacanthus nutans Induced Reactive Oxygen Species‑dependent Apoptosis and Autophagy in HCT116 Human Colorectal Cancer Cells. Pharmacognosy Magazine, 15(60), 87–97. https://doi.org/10.4103/pm.pm_299_17