PX‑12‑induced HeLa cell death is associated with oxidative stress and GSH depletion

Shin,Hye  Rim; You,Bo  Ra; Park,Woo  Hyun

doi:10.3892/ol.2013.1637

PX‑12‑induced HeLa cell death is associated with oxidative stress and GSH depletion

Authors:
- Hye Rim Shin
- Bo Ra You
- Woo Hyun Park
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Affiliations: Department of Physiology, Medical School, Research Institute for Endocrine Sciences, Chonbuk National University, Jeonju 561‑180, Republic of Korea
Published online on: October 21, 2013 https://doi.org/10.3892/ol.2013.1637
Pages: 1804-1810

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Abstract

PX‑12, as an inhibitor of thioredoxin (Trx), has antitumor activity. However, little is known about the toxicological effect of PX‑12 on cervical cancer cells. In the present study, the growth inhibitory effects of PX‑12 on HeLa cervical cancer cells in association with reactive oxygen species (ROS) and glutathione (GSH) levels were investigated. Based on MTT assays, PX‑12 inhibited the growth of HeLa cells with an IC50 value of ~7 µM at 72 h. DNA flow cytometry analysis indicated that 5 and 10 µM PX‑12 significantly induced a G2/M phase arrest of the cell cycle. PX‑12 also increased the number of dead cells and annexin V‑fluorescein isothiocyanate‑positive cells, which was accompanied by the loss of mitochondrial membrane potential. All the investigated caspase inhibitors significantly rescued certain cells from PX‑12‑induced HeLa cell death. With respect to ROS and GSH levels, PX‑12 increased ROS levels (including O2•‑) in HeLa cells and induced GSH depletion. N‑acetyl cysteine markedly reduced the levels of O2•‑ in PX‑12‑treated HeLa cells, and prevented apoptotic cell death and GSH depletion in these cells. By contrast, L‑buthionine sulfoximine intensified cell death and GSH depletion in PX‑12‑treated HeLa cells. To conclude, this is the first study to demonstrate that PX‑12 inhibits the growth of HeLa cells via G2/M phase arrest, as well as inhibiting apoptosis; the effect was associated with intracellular increases in ROS levels and GSH depletion.

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