Cadmium toxicity toward autophagy through ROS-Activated GSK-3β in mesangial cells

Sheng Hao Wang, Yung Luen Shih, Tai Chin Kuo, Wun-Chang Ko, Chwen Ming Shih

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

We previously demonstrated that cadmium (Cd) is able to induce autophagic cell death through a calcium-extracellular signal-regulated kinase pathway. Here, the object of this study is to investigate the role of glycogen synthase kinase-3β (GSK-3β) in the induction of autophagy. After treatment with Cd, MES-13 mesangial cells were determined to have undergone autophagy based on the formation of acidic vesicular organelles and autophagosomes as well as on the processing of microtubule-associated protein 1 light chain 3, using flow cytometry with acridine orange staining, electron microscopy, and immunoblot, respectively. Use of the GSK-3β inhibitor SB 216763 or the small interfering RNA technique to knockdown the expression of GSK-3β resulted in a decrease of Cd-induced autophagy. In contrast, overexpression of GSK-3β by transient transfection potentiated Cd toxicity toward the mesangial cells, suggesting that GSK-3β plays a crucial role in regulating Cd-induced autophagy. Moreover, a decrease of the phosphorylated level at Ser9 of GSK-3β was observed by immunoblot after treatment with Cd, indicating GSK-3β was activated by Cd. This phenomenon was reversed by the reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC), demonstrated that ROS might activate GSK-3β. In fact, intracellular hydrogen peroxide (H 2 O 2) was 2.6-fold elevated after 3 h of exposure to Cd. Both Cd-induced ROS bursts and autophagy were reduced by NAC and vitamin E. In summary, this study demonstrated that, in MES-13 mesangial cells, Cd-induced autophagy was mediated through the ROS-GSK-3β signaling pathway.

Original languageEnglish
Pages (from-to)124-131
Number of pages8
JournalToxicological Sciences
Volume108
Issue number1
DOIs
Publication statusPublished - 2009

Fingerprint

Glycogen Synthase Kinase 3
Mesangial Cells
Autophagy
Cadmium
Toxicity
Reactive Oxygen Species
Acetylcysteine
Acridine Orange
Microtubule-Associated Proteins
Flow cytometry
Extracellular Signal-Regulated MAP Kinases
Cell death
Vitamin E
Organelles
Hydrogen Peroxide
Electron microscopy
Small Interfering RNA
Transfection
Electron Microscopy
Flow Cytometry

Keywords

  • Autophagy
  • Cadmium
  • GSK-3β
  • Mesangial cells
  • ROS

ASJC Scopus subject areas

  • Toxicology

Cite this

Cadmium toxicity toward autophagy through ROS-Activated GSK-3β in mesangial cells. / Wang, Sheng Hao; Shih, Yung Luen; Kuo, Tai Chin; Ko, Wun-Chang; Shih, Chwen Ming.

In: Toxicological Sciences, Vol. 108, No. 1, 2009, p. 124-131.

Research output: Contribution to journalArticle

Wang, Sheng Hao ; Shih, Yung Luen ; Kuo, Tai Chin ; Ko, Wun-Chang ; Shih, Chwen Ming. / Cadmium toxicity toward autophagy through ROS-Activated GSK-3β in mesangial cells. In: Toxicological Sciences. 2009 ; Vol. 108, No. 1. pp. 124-131.
@article{0ae14379c72641b2badb6dde95af7910,
title = "Cadmium toxicity toward autophagy through ROS-Activated GSK-3β in mesangial cells",
abstract = "We previously demonstrated that cadmium (Cd) is able to induce autophagic cell death through a calcium-extracellular signal-regulated kinase pathway. Here, the object of this study is to investigate the role of glycogen synthase kinase-3β (GSK-3β) in the induction of autophagy. After treatment with Cd, MES-13 mesangial cells were determined to have undergone autophagy based on the formation of acidic vesicular organelles and autophagosomes as well as on the processing of microtubule-associated protein 1 light chain 3, using flow cytometry with acridine orange staining, electron microscopy, and immunoblot, respectively. Use of the GSK-3β inhibitor SB 216763 or the small interfering RNA technique to knockdown the expression of GSK-3β resulted in a decrease of Cd-induced autophagy. In contrast, overexpression of GSK-3β by transient transfection potentiated Cd toxicity toward the mesangial cells, suggesting that GSK-3β plays a crucial role in regulating Cd-induced autophagy. Moreover, a decrease of the phosphorylated level at Ser9 of GSK-3β was observed by immunoblot after treatment with Cd, indicating GSK-3β was activated by Cd. This phenomenon was reversed by the reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC), demonstrated that ROS might activate GSK-3β. In fact, intracellular hydrogen peroxide (H 2 O 2) was 2.6-fold elevated after 3 h of exposure to Cd. Both Cd-induced ROS bursts and autophagy were reduced by NAC and vitamin E. In summary, this study demonstrated that, in MES-13 mesangial cells, Cd-induced autophagy was mediated through the ROS-GSK-3β signaling pathway.",
keywords = "Autophagy, Cadmium, GSK-3β, Mesangial cells, ROS",
author = "Wang, {Sheng Hao} and Shih, {Yung Luen} and Kuo, {Tai Chin} and Wun-Chang Ko and Shih, {Chwen Ming}",
year = "2009",
doi = "10.1093/toxsci/kfn266",
language = "English",
volume = "108",
pages = "124--131",
journal = "Toxicological Sciences",
issn = "1096-6080",
publisher = "Oxford University Press",
number = "1",

}

TY - JOUR

T1 - Cadmium toxicity toward autophagy through ROS-Activated GSK-3β in mesangial cells

AU - Wang, Sheng Hao

AU - Shih, Yung Luen

AU - Kuo, Tai Chin

AU - Ko, Wun-Chang

AU - Shih, Chwen Ming

PY - 2009

Y1 - 2009

N2 - We previously demonstrated that cadmium (Cd) is able to induce autophagic cell death through a calcium-extracellular signal-regulated kinase pathway. Here, the object of this study is to investigate the role of glycogen synthase kinase-3β (GSK-3β) in the induction of autophagy. After treatment with Cd, MES-13 mesangial cells were determined to have undergone autophagy based on the formation of acidic vesicular organelles and autophagosomes as well as on the processing of microtubule-associated protein 1 light chain 3, using flow cytometry with acridine orange staining, electron microscopy, and immunoblot, respectively. Use of the GSK-3β inhibitor SB 216763 or the small interfering RNA technique to knockdown the expression of GSK-3β resulted in a decrease of Cd-induced autophagy. In contrast, overexpression of GSK-3β by transient transfection potentiated Cd toxicity toward the mesangial cells, suggesting that GSK-3β plays a crucial role in regulating Cd-induced autophagy. Moreover, a decrease of the phosphorylated level at Ser9 of GSK-3β was observed by immunoblot after treatment with Cd, indicating GSK-3β was activated by Cd. This phenomenon was reversed by the reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC), demonstrated that ROS might activate GSK-3β. In fact, intracellular hydrogen peroxide (H 2 O 2) was 2.6-fold elevated after 3 h of exposure to Cd. Both Cd-induced ROS bursts and autophagy were reduced by NAC and vitamin E. In summary, this study demonstrated that, in MES-13 mesangial cells, Cd-induced autophagy was mediated through the ROS-GSK-3β signaling pathway.

AB - We previously demonstrated that cadmium (Cd) is able to induce autophagic cell death through a calcium-extracellular signal-regulated kinase pathway. Here, the object of this study is to investigate the role of glycogen synthase kinase-3β (GSK-3β) in the induction of autophagy. After treatment with Cd, MES-13 mesangial cells were determined to have undergone autophagy based on the formation of acidic vesicular organelles and autophagosomes as well as on the processing of microtubule-associated protein 1 light chain 3, using flow cytometry with acridine orange staining, electron microscopy, and immunoblot, respectively. Use of the GSK-3β inhibitor SB 216763 or the small interfering RNA technique to knockdown the expression of GSK-3β resulted in a decrease of Cd-induced autophagy. In contrast, overexpression of GSK-3β by transient transfection potentiated Cd toxicity toward the mesangial cells, suggesting that GSK-3β plays a crucial role in regulating Cd-induced autophagy. Moreover, a decrease of the phosphorylated level at Ser9 of GSK-3β was observed by immunoblot after treatment with Cd, indicating GSK-3β was activated by Cd. This phenomenon was reversed by the reactive oxygen species (ROS) scavenger N-acetylcysteine (NAC), demonstrated that ROS might activate GSK-3β. In fact, intracellular hydrogen peroxide (H 2 O 2) was 2.6-fold elevated after 3 h of exposure to Cd. Both Cd-induced ROS bursts and autophagy were reduced by NAC and vitamin E. In summary, this study demonstrated that, in MES-13 mesangial cells, Cd-induced autophagy was mediated through the ROS-GSK-3β signaling pathway.

KW - Autophagy

KW - Cadmium

KW - GSK-3β

KW - Mesangial cells

KW - ROS

UR - http://www.scopus.com/inward/record.url?scp=61349090583&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=61349090583&partnerID=8YFLogxK

U2 - 10.1093/toxsci/kfn266

DO - 10.1093/toxsci/kfn266

M3 - Article

C2 - 19126599

AN - SCOPUS:61349090583

VL - 108

SP - 124

EP - 131

JO - Toxicological Sciences

JF - Toxicological Sciences

SN - 1096-6080

IS - 1

ER -