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Tumor Biology

, Volume 36, Issue 3, pp 2087–2095 | Cite as

PX-12 induces apoptosis in Calu-6 cells in an oxidative stress-dependent manner

  • Bo Ra You
  • Hye Rim Shin
  • Bo Ram Han
  • Woo Hyun Park
Research Article

Abstract

PX-12 (1-methylpropyl 2-imidazolyl disulfide) as a thioredoxin (Trx) inhibitor has an anti-tumor effect. However, there is no report about the toxicological effect of PX-12 on lung cancer cells. Here, we investigated the anti-growth effects of PX-12 on Calu-6 lung cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH) levels. PX-12 induced the growth inhibition of Calu-6 cells with IC50 of nearly 3 μM at 72 h. In contrast, PX-12 did not affect the growth of human small airway epithelial cells (HSAECs). Cell cycle distribution analysis indicated that PX-12 significantly induced a G2/M phase arrest in Calu-6 cells. PX-12 also increased the number of annexin V-FITC-positive cells in Calu-6 cells. All the tested caspase inhibitors markedly prevented Calu-6 cell death induced by PX-12. With regard to ROS and GSH levels, PX-12 increased ROS levels containing O2 ·− in Calu-6 cells and induced the depletion of GSH. N-acetyl cysteine (NAC), which is a well-known antioxidant, significantly reduced O2 ·− level in PX-12-treated Calu-6 cells and prevented apoptosis and GSH depletion in these cells. In conclusion, it is the first report that PX-12 inhibited the growth of Calu-6 cells via a G2/M phase arrest as well as apoptosis, which effect was related to the intracellular increases in ROS levels.

Keywords

PX-12 Reactive oxygen species Thioredoxin Apoptosis Lung cancer 

Abbreviations

ROS

Reactive oxygen species

Trx

Thioredoxin

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

NAC

N-Acetyl cysteine

MMP (ΔΨm)

Mitochondrial membrane potential

MTT

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

FITC

Fluorescein isothiocyanate

PI

Propidium iodide

H2DCFDA

2′,7′-dichlorodihydrofluorescein diacetate

DHE

Dihydroethidium

CMFDA

5-Chloromethylfluorescein diacetate

Notes

Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea Government (MSIP) (No. 2008–0062279) and supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2013006279). This paper was supported by research funds of Chonbuk National University in 2014.

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Bo Ra You
    • 1
  • Hye Rim Shin
    • 1
  • Bo Ram Han
    • 1
  • Woo Hyun Park
    • 1
    • 2
  1. 1.Department of Physiology, Medical School, Research Institute for Endocrine SciencesChonbuk National UniversityJeonJuRepublic of Korea
  2. 2.Department of Physiology, Medical SchoolChonbuk National UniversityJeonJuRepublic of Korea

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