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http://dx.doi.org/10.1016/j.jgr.2016.04.003

Induction of Forkhead Class box O3a and apoptosis by a standardized ginsenoside formulation, KG-135, is potentiated by autophagy blockade in A549 human lung cancer cells  

Yao, Chih-Jung (Cancer Center, Wan Fang Hospital, Taipei Medical University)
Chow, Jyh-Ming (Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University)
Chuang, Shuang-En (National Institute of Cancer Research, National Health Research Institutes)
Chang, Chia-Lun (Division of Hematology and Medical Oncology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University)
Yan, Ming-De (Cancer Center, Wan Fang Hospital, Taipei Medical University)
Lee, Hsin-Lun (Translational Medicine, Taipei Medical University-Academia Sinica)
Lai, I-Chun (Department of Oncology, Taipei Veterans General Hospital)
Lin, Pei-Chun (Cancer Center, Wan Fang Hospital, Taipei Medical University)
Lai, Gi-Ming (Cancer Center, Wan Fang Hospital, Taipei Medical University)
Publication Information
Journal of Ginseng Research / v.41, no.3, 2017 , pp. 247-256 More about this Journal
Abstract
Background: KG-135, a standardized formulation enriched with Rk1, Rg3, and Rg5 ginsenosides, has been shown to inhibit various types of cancer cells; however, the underlying mechanisms are not fully understood. In this study, we explored its effects in A549 human lung cancer cells to investigate the induction of Forkhead Class box O3a (FOXO3a) and autophagy. Methods: Cell viability was determined by sulforhodamine B staining. Apoptosis and cell cycle distribution were analyzed using flow cytometry. The changes of protein levels were determined using Western blot analysis. Autophagy induction was monitored by the formation of acidic vesicular organelles stained with acridine orange. Results: KG-135 effectively arrested the cells in G1 phase with limited apoptosis. Accordingly, a decrease of cyclin-dependent kinase-4, cyclin-dependent kinase-6, cyclin D1, and phospho-retinoblastoma protein, and an increase of p27 and p18 proteins were observed. Intriguingly, KG-135 increased the tumor suppressor FOXO3a and induced the accumulation of autophagy hallmark LC3-II and acidic vesicular organelles without an increase of the upstream marker Beclin-1. Unconventionally, the autophagy adaptor protein p62 (sequestosome 1) was increased rather than decreased. Blockade of autophagy by hydroxychloroquine dramatically potentiated KG-135-induced FOXO3a and its downstream (FasL) ligand accompanied by the cleavage of caspase-8. Meanwhile, the decrease of Bcl-2 and survivin, as well as the cleavage of caspase-9, were also drastically enhanced, resulting in massive apoptosis. Conclusion: Besides arresting the cells in G1 phase, KG-135 increased FOXO3a and induced an unconventional autophagy in A549 cells. Both the KG-135-activated extrinsic FOXO3a/FasL/caspase-8 and intrinsic caspase-9 apoptotic pathways were potentiated by blockade of autophagy. Combination of KG-135 and autophagy inhibitor may be a novel strategy as an integrative treatment for cancers.
Keywords
autophagy; FOXO3a; ginsenoside; KG-135; lung cancer;
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