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The effects of MAPK inhibitors on antimycin A-treated Calu-6 lung cancer cells in relation to cell growth, reactive oxygen species, and glutathione

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Abstract

Antimycin A (AMA) inhibits succinate oxidase, NADH oxidase, and mitochondrial electron transport chain between cytochrome b and c. We recently demonstrated that AMA inhibited the growth of Calu-6 lung cancer cells through apoptosis. Here, we investigated the effects of AMA and/or MAPK inhibitors on Calu-6 lung cancer cells in relation to cell growth, cell death, reactive oxygen species (ROS), and GSH levels. Treatment with AMA inhibited the growth of Calu-6 cells at 72 h. AMA-induced apoptosis was accompanied by the loss of mitochondrial membrane potential (MMP; ΔΨm). While ROS were decreased in AMA-treated Calu-6 cells, O •−2 among ROS was increased. AMA also induced GSH depletion in Calu-6 cells. Treatment with MEK inhibitor intensified cell death, MMP (ΔΨm) loss, and GSH depletion in AMA-treated Calu-6 cells. JNK inhibitor also increased cell death, MMP (ΔΨm) loss, and ROS levels in these cells. Treatment with p38 inhibitor magnified cell growth inhibition by AMA and increased cell death, MMP (ΔΨm) loss, ROS level, and GSH depletion in AMA-treated cells. Conclusively, all the MAPK inhibitors slightly intensified cell death in AMA-treated Calu-6 cells. The changes of ROS and GSH by AMA and/or MAPK inhibitors were in part involved in cell growth and death in Calu-6 cells.

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Abbreviations

AMA:

Antimycin A

ROS:

Reactive oxygen species

MAPK:

Mitogen-activated protein kinase

MEK:

MAP kinase or ERK kinase

ERK:

Extracellular signal-regulated kinase

JNK:

c-Jun N-terminal kinase

SOD:

Superoxide dismutase

MMP (ΔΨm):

Mitochondrial membrane potential

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

H2DCFDA:

2′7′-dichlorodihydrofluorescein diacetate

DHE:

Dihydroethidium

GSH:

Glutathione

CMFDA:

5-Chloromethylfluorescein diacetate

MTT:

3-(45-Dimethylthiazol-2-yl)-25-diphenyltetrazolium bromide

PI:

Propidium iodide

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Acknowledgments

This research was supported by a grant of the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A084194).

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  1. Department of Physiology, Medical School, Centers for Healthcare Technology Development, Institute for Medical Sciences, Chonbuk National University, JeonJu, 561-180, Republic of Korea

    Yong Hwan Han & Woo Hyun Park

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  1. Yong Hwan Han
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Correspondence to Woo Hyun Park.

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Han, Y.H., Park, W.H. The effects of MAPK inhibitors on antimycin A-treated Calu-6 lung cancer cells in relation to cell growth, reactive oxygen species, and glutathione. Mol Cell Biochem 333, 211–219 (2010). https://doi.org/10.1007/s11010-009-0222-2

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  • DOI: https://doi.org/10.1007/s11010-009-0222-2

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