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Antiapoptotic effects of caspase inhibitors on H2O2-treated lung cancer cells concerning oxidative stress and GSH

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Abstract

Exogenous hydrogen peroxide (H2O2) induces oxidative stress and apoptosis in cancer cells. This study evaluated the antiapoptotic effects of pan-caspase and caspase-3, -8, or -9 inhibitors on H2O2-treated Calu-6 and A549 lung cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH). Treatment with 50–500 μM H2O2 inhibited the growth of Calu-6 and A549 cells at 24 h and induced apoptosis in these cells. All the tested caspase inhibitors significantly prevented cell death in H2O2-treated lung cancer cells. H2O2 increased intracellular ROS levels, including that of O ·−2 , at 1 and 24 h. It also increased the activity of catalase but decreased the activity of SOD. In addition, H2O2 triggered GSH deletion in Calu-6 and A549 cells at 24 h. It reduced GSH levels in Calu-6 cells at 1 h but increased them at 24 h. Caspase inhibitors decreased O ·−2 levels in H2O2-treated Calu-6 cells at 1 h and these inhibitors decreased ROS levels, including that of O ·−2 , in H2O2-treated A549 cells at 24 h. Caspase inhibitors partially attenuated GSH depletion in H2O2-treated A549 cells and increased GSH levels in these cells at 24 h. However, the inhibitors did not affect GSH deletion and levels in Calu-6 cells at 24 h. In conclusion, H2O2 induced caspase-dependent apoptosis in Calu-6 and A549 cells, which was accompanied by increases in ROS and GSH depletion. The antiapoptotic effects of caspase inhibitors were somewhat related to the suppression of H2O2-induced oxidative stress and GSH depletion.

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Abbreviations

H2O2 :

Hydrogen peroxide

ROS:

Reactive oxygen species

GSH:

Glutathione

Z-VAD-FMK:

Benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone

Z-DEVD-FMK:

Benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone

Z-IETD-FMK:

Benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethylketone

Z-LEHD-FMK:

Benzyloxycarbonyl-Leu-Glu-His-Asp-fluoromethylketone

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

SOD:

Superoxide dismutase

H2DCFDA:

2′,7′-Dichlorodihydrofluorescein diacetate

DHE:

Dihydroethidium

CMFDA:

5-Chloromethylfluorescein diacetate

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Acknowledgements

This study was supported by a Grant from the National Research Foundation of Korea (NRF) funded by the Korean government (MSIP; No 2016R1A2B4007773) and supported by “Research Base Construction Fund Support Program” funded by Chonbuk National University in 2016.

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  1. Department of Physiology, Medical School, Research Institute for Endocrine Sciences, Chonbuk National University, 20 Geonji-ro, Deokjin-gu, Jeonju, Jeollabuk-do, 54907, Republic of Korea

    Woo Hyun Park

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Correspondence to Woo Hyun Park.

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Park, W.H. Antiapoptotic effects of caspase inhibitors on H2O2-treated lung cancer cells concerning oxidative stress and GSH. Mol Cell Biochem 441, 125–134 (2018). https://doi.org/10.1007/s11010-017-3179-6

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  • DOI: https://doi.org/10.1007/s11010-017-3179-6

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