Preprint Article Version 1 NOT YET PEER-REVIEWED

The Apoptosis Induction of Oleandrin on Osteosarcoma Cells through Regulating Mitochondrial- and Death Receptor-Dependent Apoptotic Pathways in Vitro

  1. Department of Orthopedics, Peking University Third Hospital, No. 49, North Garden Street, Haidian District, Beijing 100191, China
  2. The Center for Pain medicine, Peking University Third Hospital, No. 49, North Garden Street, Haidian District, Beijing 100191, China
  3. Department of Anesthesiology, Peking University Third Hospital, No. 49, North Garden Street, Haidian District, Beijing 100191, China
Version 1 : Received: 21 August 2016 / Approved: 22 August 2016 / Online: 22 August 2016 (05:35:06 CEST)

A peer-reviewed article of this Preprint also exists.

Ma, Y.; Zhu, B.; Yong, L.; Song, C.; Liu, X.; Yu, H.; Wang, P.; Liu, Z.; Liu, X. Regulation of Intrinsic and Extrinsic Apoptotic Pathways in Osteosarcoma Cells Following Oleandrin Treatment. Int. J. Mol. Sci. 2016, 17, 1950. Ma, Y.; Zhu, B.; Yong, L.; Song, C.; Liu, X.; Yu, H.; Wang, P.; Liu, Z.; Liu, X. Regulation of Intrinsic and Extrinsic Apoptotic Pathways in Osteosarcoma Cells Following Oleandrin Treatment. Int. J. Mol. Sci. 2016, 17, 1950.

Journal reference: Int. J. Mol. Sci. 2016, 17, 1950
DOI: 10.3390/ijms17111950

Abstract

Our previous study has found the anti-tumor activity of oleandrin in osteosarcoma cells in vitro, but the signal transduction process of cell apoptosis induced by oleandrin is uncertain, which is explored in this study. Fluorescence staining and flow cytometry (FCM) was performed to detect the cell apoptosis, intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (MMP). Caspase-3 activity was detected using a commercial kit. The protein expression of cytoplasmic cytochrome c, mitochondrial cytochrome c, bcl-2, bax, caspase-9, Fas, FasL, caspase-8 and caspase-3 was detected using western blot. A pan-caspase inhibitor, z-VAD-fmk, was applied to block the apoptotic pathway and the apoptosis status were re-tested. We found that oleandrin significantly induced the increased apoptosis of U2OS cells. Meanwhile, the intracellular ROS was elevated, but the MMP decreased. The cytochrome c in mitochondria was notably decreased but increased in cytoplasm. The caspase-3 activity was also enhanced with the increase of drug concentration and treatment time. Oleandrin also down-regulated the level of bcl-2, but remarkably up-regulated the expression of bax, cleaved caspase-9, Fas, FasL, cleaved caspase-8 and cleaved caspase-3. Furthermore, the pre-treatment with z-VAD-fmk almost completely reverted the oleandrin-induced apoptosis. The results suggested that oleandrin induces the apoptosis of osteosarcoma cells via mitochondrial- and death receptor-dependent pathways.

Subject Areas

oleandrin; osteosarcoma; apoptosis; caspases

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