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Deoxypodophyllotoxin induces cytoprotective autophagy against apoptosis via inhibition of PI3K/AKT/mTOR pathway in osteosarcoma U2OS cells

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Abstract

Background

A natural compound deoxypodophyllotoxin (DPT) possesses potent anti-proliferative and anti-tumor properties on several cancer types. It triggers cell cycle arrest followed by apoptosis through various cellular processes. However, it is limited to the action mechanism of DPT-mediated cell death modes via apoptosis and autophagy.

Methods

Cell viability assay, morphological changes, annexin-V/propidium iodide (PI) assay, reactive oxygen species (ROS), acridine orange staining, and Western blot analyses were evaluated.

Results

We demonstrated that DPT induced both apoptosis and autophagy via production of mitochondrial reactive oxygen species (ROS). DPT suppressed the PI3 K/AKT/mTOR signaling cascades to lead autophagy process, resulting from conversion of light chain 3-I (LC3-I) into LC3-II and acidic vesicular organelles (AVOs) formation. Even if DPT-induced ROS were occurred in both apoptosis and autophagy, inhibition of ROS generation enhanced cell viability. Otherwise, 3-methyladeine (3-MA) impeding on autophagy accelerated an apoptotic response caused by DPT. Therefore, these findings suggest that DPT triggers cytoprotective autophagy against cytotoxic apoptosis.

Conclusion

Autophagy is required for cell survival by inhibition of apoptosis through down-regulation of PI3K/AKT/mTOR pathway against DPT-induced apoptosis in U2OS cells.

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Author notes

  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology & Immunology, Pusan National University School of Medicine, Yangsan, Republic of Korea

    Sang-Hun Kim, Sun-Nyoung Yu, Sul-Gi Park, Young-Wook Kim, Hyo-Won Nam & Soon-Cheol Ahn

  2. Department of Orthopedic Surgery, Hana Hospital, Changwon, Republic of Korea

    Kyo-Min Son

  3. Department of Herbal Formula, Medical Research Center (MRC-GHF), College of Oriental Medicine, Daegu Haany University, Gyeongsan, Republic of Korea

    Kwang-Youn Kim

  4. Department of Orthopedic Surgery, Pusan National University School of Medicine, Busan, Republic of Korea

    Jeung-Tak Suh

  5. Genome Instability Research Center, Ajou University School of Medicine, Suwon, Republic of Korea

    Jae-Hoon Ji

  6. Immunoregulatory Therapeutics Group in Brain Busan 21 Project, Pusan National University, Yangsan, Republic of Korea

    Soon-Cheol Ahn

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  1. Sang-Hun Kim
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  2. Kyo-Min Son
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Correspondence to Jae-Hoon Ji or Soon-Cheol Ahn.

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Kim, SH., Son, KM., Kim, KY. et al. Deoxypodophyllotoxin induces cytoprotective autophagy against apoptosis via inhibition of PI3K/AKT/mTOR pathway in osteosarcoma U2OS cells. Pharmacol. Rep 69, 878–884 (2017). https://doi.org/10.1016/j.pharep.2017.04.007

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  • DOI: https://doi.org/10.1016/j.pharep.2017.04.007

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