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Reversine Induces Caspase-Dependent Apoptosis of Human Osteosarcoma Cells Through Extrinsic and Intrinsic Apoptotic Signaling Pathways

Reversine Induces Caspase-Dependent Apoptosis of Human Osteosarcoma Cells Through Extrinsic and Intrinsic Apoptotic Signaling Pathways

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, Do Kyung Kim

Genes Genomics. 2019 Jun;41(6):657-665.

PMID: 30953339

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP656820325	Reversine		656820-32-5	Price

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