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Ouabain Induces Apoptotic Cell Death Through Caspase- and Mitochondria-dependent Pathways in Human Osteosarcoma U-2

Wen-Hsiang Chou1,2,3, Ko-Lin Liu4, Yung-Luen Shih4,5

  • 1School of Medicine, National Defense Medical Center, Taipei, Taiwan, R.O.C.

Anticancer Research
|December 27, 2017
PubMed
Abstract

Insights

Ouabain, a plant-derived substance, induces programmed cell death (apoptosis) in human osteosarcoma U-2 OS cells. This study details ouabain

Area of Science:

  • Biochemistry and Molecular Biology
  • Cell Biology
  • Pharmacology

Background:

  • Ouabain, a cardiac glycoside, exhibits Na+/K+-ATPase inhibitory properties.
  • Previous research indicates ouabain possesses anti-cancer activity against various human cancer cells.
  • This study is the first to investigate ouabain's impact on apoptotic cell death in human osteosarcoma U-2 OS cells.

Purpose of the Study:

  • To investigate the effect of ouabain on inducing apoptotic cell death in human osteosarcoma U-2 OS cells.
  • To elucidate the mechanisms underlying ouabain-induced apoptosis in U-2 OS cells.

Main Methods:

  • Cell viability, cell cycle, reactive oxygen species (ROS), intracellular calcium (Ca2+), and mitochondrial membrane potential (MMP) were assessed using flow cytometry.
  • Morphological changes were observed via contrast-phase microscopy.
  • Apoptosis-associated protein levels, including Bax, Bcl-2, cytochrome c, apoptosis-inducing factor (AIF), and endonuclease G (Endo G), were analyzed by Western blot.

Main Results:

  • Ouabain significantly reduced viable U-2 OS cells and induced morphological changes in a time-dependent manner.
  • Ouabain altered the cell cycle distribution, decreasing the G0/G1 phase and increasing S and G2/M phases.
  • Ouabain modulated intracellular signaling pathways, increasing caspase-3, -8, and -9 activities, altering Bax/Bcl-2 expression, and promoting the release of apoptosis-related proteins (cytochrome c, AIF, Endo G) via both caspase-dependent and -independent pathways.

Conclusions:

  • Ouabain demonstrates significant cytotoxic effects on U-2 OS cells in vitro.
  • Ouabain-induced cytotoxicity is mediated, at least in part, by the induction of apoptosis.
  • These findings provide crucial insights into the anti-cancer potential of ouabain against osteosarcoma.
Keywords:
Ouabainapoptosiscaspase-dependent pathwayshuman osteosarcoma U-2 OS cellsmitochondria-dependent pathways

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