Involvement of reactive oxygen species in arsenite-induced downregulation of phospholipid hydroperoxide glutathione peroxidase in human epidermoid carcinoma A431 cells

Huei Sheng Huang, Wen Chang Chang, Ching Jiunn Chen

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is unique in the substrate specificity among the glutathione peroxidase family because it can interact with lipophilic substrates, including the peroxidized phospholipids and cholesterol, and reduce these hydroperoxide to hydroxide compounds. However, what kinds of ligand can regulate the PHGPx expression is still unknown. In the present study, we found that sodium arsenite induced downregulation of mRNA, protein expression, and enzyme activity of PHGPx in time- and dose-dependent manners. At the same time, it upregulated mRNA and protein expression of p21WAF1/CIP1. With the aid of agarose gel electrophoresis, and propidium iodide and annexin-V staining, we found that treatment of 30 μM sodium arsenite for 24 h induced apoptosis in human epidermoid carcinoma A431 cells and EA.hy926 cells. An increase of intracellular peroxide levels was measured by flow cytometry using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) after treatment of arsenite. Overexpression of PHGPx prevented arsenite-induced increase of intracellular peroxide levels, downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells. N-Acetyl-L-cysteine also significantly prevented arsenite-induced effects in A431 cells. Therefore, we concluded that reactive oxygen species were involved in arsenite-induced downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells.

Original languageEnglish
Pages (from-to)864-873
Number of pages10
JournalFree Radical Biology and Medicine
Volume33
Issue number6
DOIs
Publication statusPublished - Sep 15 2002
Externally publishedYes

Fingerprint

phospholipid-hydroperoxide glutathione peroxidase
Squamous Cell Carcinoma
Reactive Oxygen Species
Down-Regulation
Peroxides
Apoptosis
Up-Regulation
Messenger RNA
Flow cytometry
Agar Gel Electrophoresis
Propidium
Annexin A5
Acetylcysteine
Enzyme activity
Substrates
Glutathione Peroxidase
Substrate Specificity
Electrophoresis
Sepharose
arsenite

Keywords

  • Arsenite
  • Free radicals
  • P21
  • PHGPx
  • ROS

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medicine(all)
  • Toxicology

Cite this

@article{b7c6223088f344c5b77560da585139dd,
title = "Involvement of reactive oxygen species in arsenite-induced downregulation of phospholipid hydroperoxide glutathione peroxidase in human epidermoid carcinoma A431 cells",
abstract = "Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is unique in the substrate specificity among the glutathione peroxidase family because it can interact with lipophilic substrates, including the peroxidized phospholipids and cholesterol, and reduce these hydroperoxide to hydroxide compounds. However, what kinds of ligand can regulate the PHGPx expression is still unknown. In the present study, we found that sodium arsenite induced downregulation of mRNA, protein expression, and enzyme activity of PHGPx in time- and dose-dependent manners. At the same time, it upregulated mRNA and protein expression of p21WAF1/CIP1. With the aid of agarose gel electrophoresis, and propidium iodide and annexin-V staining, we found that treatment of 30 μM sodium arsenite for 24 h induced apoptosis in human epidermoid carcinoma A431 cells and EA.hy926 cells. An increase of intracellular peroxide levels was measured by flow cytometry using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) after treatment of arsenite. Overexpression of PHGPx prevented arsenite-induced increase of intracellular peroxide levels, downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells. N-Acetyl-L-cysteine also significantly prevented arsenite-induced effects in A431 cells. Therefore, we concluded that reactive oxygen species were involved in arsenite-induced downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells.",
keywords = "Arsenite, Free radicals, P21, PHGPx, ROS",
author = "Huang, {Huei Sheng} and Chang, {Wen Chang} and Chen, {Ching Jiunn}",
year = "2002",
month = "9",
day = "15",
doi = "10.1016/S0891-5849(02)00983-8",
language = "English",
volume = "33",
pages = "864--873",
journal = "Free Radical Biology and Medicine",
issn = "0891-5849",
publisher = "Elsevier Inc.",
number = "6",

}

TY - JOUR

T1 - Involvement of reactive oxygen species in arsenite-induced downregulation of phospholipid hydroperoxide glutathione peroxidase in human epidermoid carcinoma A431 cells

AU - Huang, Huei Sheng

AU - Chang, Wen Chang

AU - Chen, Ching Jiunn

PY - 2002/9/15

Y1 - 2002/9/15

N2 - Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is unique in the substrate specificity among the glutathione peroxidase family because it can interact with lipophilic substrates, including the peroxidized phospholipids and cholesterol, and reduce these hydroperoxide to hydroxide compounds. However, what kinds of ligand can regulate the PHGPx expression is still unknown. In the present study, we found that sodium arsenite induced downregulation of mRNA, protein expression, and enzyme activity of PHGPx in time- and dose-dependent manners. At the same time, it upregulated mRNA and protein expression of p21WAF1/CIP1. With the aid of agarose gel electrophoresis, and propidium iodide and annexin-V staining, we found that treatment of 30 μM sodium arsenite for 24 h induced apoptosis in human epidermoid carcinoma A431 cells and EA.hy926 cells. An increase of intracellular peroxide levels was measured by flow cytometry using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) after treatment of arsenite. Overexpression of PHGPx prevented arsenite-induced increase of intracellular peroxide levels, downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells. N-Acetyl-L-cysteine also significantly prevented arsenite-induced effects in A431 cells. Therefore, we concluded that reactive oxygen species were involved in arsenite-induced downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells.

AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is unique in the substrate specificity among the glutathione peroxidase family because it can interact with lipophilic substrates, including the peroxidized phospholipids and cholesterol, and reduce these hydroperoxide to hydroxide compounds. However, what kinds of ligand can regulate the PHGPx expression is still unknown. In the present study, we found that sodium arsenite induced downregulation of mRNA, protein expression, and enzyme activity of PHGPx in time- and dose-dependent manners. At the same time, it upregulated mRNA and protein expression of p21WAF1/CIP1. With the aid of agarose gel electrophoresis, and propidium iodide and annexin-V staining, we found that treatment of 30 μM sodium arsenite for 24 h induced apoptosis in human epidermoid carcinoma A431 cells and EA.hy926 cells. An increase of intracellular peroxide levels was measured by flow cytometry using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) after treatment of arsenite. Overexpression of PHGPx prevented arsenite-induced increase of intracellular peroxide levels, downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells. N-Acetyl-L-cysteine also significantly prevented arsenite-induced effects in A431 cells. Therefore, we concluded that reactive oxygen species were involved in arsenite-induced downregulation of PHGPx, upregulation of p21WAF1/CIP1, and apoptosis in A431 cells.

KW - Arsenite

KW - Free radicals

KW - P21

KW - PHGPx

KW - ROS

UR - http://www.scopus.com/inward/record.url?scp=0037105320&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037105320&partnerID=8YFLogxK

U2 - 10.1016/S0891-5849(02)00983-8

DO - 10.1016/S0891-5849(02)00983-8

M3 - Article

VL - 33

SP - 864

EP - 873

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 6

ER -