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Reversine induces caspase-dependent apoptosis of human osteosarcoma cells through extrinsic and intrinsic apoptotic signaling pathways

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Abstract

Background

The 2-(4-morpholinoanilino)-6-cyclohexylaminopurine (reversine) acts as a chemopreventive agent and induces apoptotic cell death in various cancer cells. However, the anticancer effects of reversine on osteosarcoma cells are not clearly established.

Objective

The purpose of this study was to investigate the effect of reversine on cell proliferation and induction of apoptosis in human osteosarcoma cells.

Methods

Cell viability assay, histological analysis, DAPI staining, caspase activation analysis, flow cytometric analysis and immunoblotting were carried out in MG-63 osteosarcoma cells.

Results

Reversine inhibited the growth of cells in a dose-dependent manner and induced nuclear condensation and fragmentation. Reversine-treated cells showed caspase-3/7 activation and increased apoptosis versus control cells. FasL, a death ligand associated with extrinsic apoptotic signaling pathways, was significantly up-regulated by reversine treatment. Moreover, the caspase-8, a part of the extrinsic apoptotic pathway, was activated by reversine treatments. Expressions of anti-apoptotic factors such as Bcl-2 and Bcl-xL, components of the mitochondria dependent intrinsic apoptosis pathway, significantly decreased following reversine treatment. The expressions of pro-apoptotic factors such as BAX, BAD and caspase-9 increased by reversine treatments. In addition, reversine activated caspase-3 and Poly (ADP-ribose) polymerase (PARP) to induce cell death. The Z-VAD-fmk significantly inhibited cell death through the suppression of caspase-3 expression in MG-63 cells treated with reversine.

Conclusion

These results suggest that the reversine may inhibit cell proliferation and induce apoptotic cell death in MG-63 osteosarcoma cells through both the mitochondria-mediated intrinsic pathway and the death receptor-mediated extrinsic pathway, and may have potential properties for the discovery of anti-cancer agents.

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Authors and Affiliations

  1. Institute of Dental Science, Chosun University, Gwangju, 61452, Republic of Korea

    Jae-Sung Kim, In-A Cho, Kyeong-Rok Kang, HyangI Lim, Tae-Hyeon Kim, Sun-Kyoung Yu, Heung-Joong Kim, Seul Ah Lee, Sung Min Moon, Chun Sung Kim & Do Kyung Kim

  2. Department of Biomedical Science, Chosun University, Gwangju, 61452, Republic of Korea

    Hong Sung Chun

  3. Department of Oral Physiology, School of Dentistry, Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju, 61452, Republic of Korea

    Do Kyung Kim

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  1. Jae-Sung Kim
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  2. In-A Cho
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Correspondence to Do Kyung Kim.

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Conflict of interest

Jae-Sung Kim, In-A Cho, Kyeong-Rok Kang, HyangI Lim, Tae-Hyeon Kim, Sun-Kyoung Yu, Heung-Joong Kim, Seul Ah Lee, Sung Min Moon, Hong Sung Chun, Chun Sung Kim and Do Kyung Kim declare that they have no conflict of interest.

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Kim, JS., Cho, IA., Kang, KR. et al. Reversine induces caspase-dependent apoptosis of human osteosarcoma cells through extrinsic and intrinsic apoptotic signaling pathways. Genes Genom 41, 657–665 (2019). https://doi.org/10.1007/s13258-019-00790-1

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