GSK3β and endoplasmic reticulum stress mediate rotenone-induced death of SK-N-MC neuroblastoma cells

Yuan Yuan Chen; Gang Chen; Zhiqin Fan; Jia Luo; Zun Ji Ke

doi:10.1016/j.bcp.2008.04.010

GSK3β and endoplasmic reticulum stress mediate rotenone-induced death of SK-N-MC neuroblastoma cells

Yuan Yuan Chen, Gang Chen, Zhiqin Fan, Jia Luo, Zun Ji Ke

Pharmacology and Nutritional Sciences

Research output: Contribution to journal › Article › peer-review

51 Scopus citations

Abstract

Rotenone, an environmental toxin that inhibits mitochondrial complex I, has been used to induce experimental Parkinsonism in animals and cell cultures. We investigated the mechanism underlying rotenone-induced death of SK-N-MC neuroblastoma cells. Rotenone-induced cell death preceded intracellular accumulation of reactive oxygen species, and antioxidants failed to protect cells, indicating that oxidative stress was minimally involved in rotenone-induced death of SK-N-MC cells. Glycogen synthase kinase 3β (GSK3β), a multifunctional serine/threonine kinase, has been implicated in the pathogenesis of neurodegeneration. We showed that rotenone activated GSK3β by enhancing its phosphorylation at tyrosine 216 while inhibiting phosphorylation at serine 9. Inhibitors of GSK3β and dominant negative (kinase deficient) GSK3β partially protected SK-N-MC cells against rotenone cytotoxicity. Rotenone also induced endoplasmic reticulum (ER) stress which was evident by an increase in phosphorylation of PERK, PKR, and eIF2α as well as the expression of GRP78. Rotenone had a modest effect on the expression of CHOP. An eIF2α siRNA significantly reduced rotenone cytotoxicity. ER stress was experimentally induced by tunicamycin and thapsigargin, but tunicamycin/thapsigargin did not activate GSK3β in SK-N-MC cells. Down-regulation of eIF2α also offered partial protection against rotenone cytotoxicity. Combined treatment of GSK3β inhibitors and eIF2α siRNA provided much greater protection than either treatment alone. Taken together, the results suggest that GSK3β activation and ER stress contribute separately to rotenone cytotoxicity.

Original language	English
Pages (from-to)	128-138
Number of pages	11
Journal	Biochemical Pharmacology
Volume	76
Issue number	1
DOIs	https://doi.org/10.1016/j.bcp.2008.04.010
State	Published - Jul 1 2008

Bibliographical note

Funding Information:
We would like to thank Ms. Kimberly A. Bower for reading this manuscript. This research was supported by grants from the Ministry of Science and Technology of China (2007CB947100), the National Natural Science Foundation of China (30470544, 30471452, and 30570580), the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-115), and Science and Technology Commission of Shanghai Municipality (Grant No. 07DJ14005). Dr Z.-J. Ke was also supported by the One Hundred Talents Program of the Chinese Academy of Sciences. Dr J. Luo was also supported by a grant from NIH/NIAAA (AA015407).

Funding

We would like to thank Ms. Kimberly A. Bower for reading this manuscript. This research was supported by grants from the Ministry of Science and Technology of China (2007CB947100), the National Natural Science Foundation of China (30470544, 30471452, and 30570580), the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-115), and Science and Technology Commission of Shanghai Municipality (Grant No. 07DJ14005). Dr Z.-J. Ke was also supported by the One Hundred Talents Program of the Chinese Academy of Sciences. Dr J. Luo was also supported by a grant from NIH/NIAAA (AA015407).

Funders	Funder number
Ministry of Science and Technology, China	2007CB947100
DBR/NIAAA/NIH	AA015407
National Institute on Alcohol Abuse and Alcoholism	R01AA015407
National Natural Science Foundation of China (NSFC)	30471452, 30470544, 30570580
Chinese Academy of Sciences	KSCX2-YW-R-115
Science and Technology Commission of Shanghai Municipality	07DJ14005

Keywords

ER stress
GSK3β
Neurodegeneration
Oxidative stress
Parkinson's disease

ASJC Scopus subject areas

Biochemistry
Pharmacology

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10.1016/j.bcp.2008.04.010

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title = "GSK3β and endoplasmic reticulum stress mediate rotenone-induced death of SK-N-MC neuroblastoma cells",

abstract = "Rotenone, an environmental toxin that inhibits mitochondrial complex I, has been used to induce experimental Parkinsonism in animals and cell cultures. We investigated the mechanism underlying rotenone-induced death of SK-N-MC neuroblastoma cells. Rotenone-induced cell death preceded intracellular accumulation of reactive oxygen species, and antioxidants failed to protect cells, indicating that oxidative stress was minimally involved in rotenone-induced death of SK-N-MC cells. Glycogen synthase kinase 3β (GSK3β), a multifunctional serine/threonine kinase, has been implicated in the pathogenesis of neurodegeneration. We showed that rotenone activated GSK3β by enhancing its phosphorylation at tyrosine 216 while inhibiting phosphorylation at serine 9. Inhibitors of GSK3β and dominant negative (kinase deficient) GSK3β partially protected SK-N-MC cells against rotenone cytotoxicity. Rotenone also induced endoplasmic reticulum (ER) stress which was evident by an increase in phosphorylation of PERK, PKR, and eIF2α as well as the expression of GRP78. Rotenone had a modest effect on the expression of CHOP. An eIF2α siRNA significantly reduced rotenone cytotoxicity. ER stress was experimentally induced by tunicamycin and thapsigargin, but tunicamycin/thapsigargin did not activate GSK3β in SK-N-MC cells. Down-regulation of eIF2α also offered partial protection against rotenone cytotoxicity. Combined treatment of GSK3β inhibitors and eIF2α siRNA provided much greater protection than either treatment alone. Taken together, the results suggest that GSK3β activation and ER stress contribute separately to rotenone cytotoxicity.",

keywords = "ER stress, GSK3β, Neurodegeneration, Oxidative stress, Parkinson's disease",

author = "Chen, {Yuan Yuan} and Gang Chen and Zhiqin Fan and Jia Luo and Ke, {Zun Ji}",

note = "Funding Information: We would like to thank Ms. Kimberly A. Bower for reading this manuscript. This research was supported by grants from the Ministry of Science and Technology of China (2007CB947100), the National Natural Science Foundation of China (30470544, 30471452, and 30570580), the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-115), and Science and Technology Commission of Shanghai Municipality (Grant No. 07DJ14005). Dr Z.-J. Ke was also supported by the One Hundred Talents Program of the Chinese Academy of Sciences. Dr J. Luo was also supported by a grant from NIH/NIAAA (AA015407).",

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AU - Chen, Gang

AU - Fan, Zhiqin

AU - Luo, Jia

AU - Ke, Zun Ji

N1 - Funding Information: We would like to thank Ms. Kimberly A. Bower for reading this manuscript. This research was supported by grants from the Ministry of Science and Technology of China (2007CB947100), the National Natural Science Foundation of China (30470544, 30471452, and 30570580), the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-115), and Science and Technology Commission of Shanghai Municipality (Grant No. 07DJ14005). Dr Z.-J. Ke was also supported by the One Hundred Talents Program of the Chinese Academy of Sciences. Dr J. Luo was also supported by a grant from NIH/NIAAA (AA015407).

PY - 2008/7/1

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N2 - Rotenone, an environmental toxin that inhibits mitochondrial complex I, has been used to induce experimental Parkinsonism in animals and cell cultures. We investigated the mechanism underlying rotenone-induced death of SK-N-MC neuroblastoma cells. Rotenone-induced cell death preceded intracellular accumulation of reactive oxygen species, and antioxidants failed to protect cells, indicating that oxidative stress was minimally involved in rotenone-induced death of SK-N-MC cells. Glycogen synthase kinase 3β (GSK3β), a multifunctional serine/threonine kinase, has been implicated in the pathogenesis of neurodegeneration. We showed that rotenone activated GSK3β by enhancing its phosphorylation at tyrosine 216 while inhibiting phosphorylation at serine 9. Inhibitors of GSK3β and dominant negative (kinase deficient) GSK3β partially protected SK-N-MC cells against rotenone cytotoxicity. Rotenone also induced endoplasmic reticulum (ER) stress which was evident by an increase in phosphorylation of PERK, PKR, and eIF2α as well as the expression of GRP78. Rotenone had a modest effect on the expression of CHOP. An eIF2α siRNA significantly reduced rotenone cytotoxicity. ER stress was experimentally induced by tunicamycin and thapsigargin, but tunicamycin/thapsigargin did not activate GSK3β in SK-N-MC cells. Down-regulation of eIF2α also offered partial protection against rotenone cytotoxicity. Combined treatment of GSK3β inhibitors and eIF2α siRNA provided much greater protection than either treatment alone. Taken together, the results suggest that GSK3β activation and ER stress contribute separately to rotenone cytotoxicity.

AB - Rotenone, an environmental toxin that inhibits mitochondrial complex I, has been used to induce experimental Parkinsonism in animals and cell cultures. We investigated the mechanism underlying rotenone-induced death of SK-N-MC neuroblastoma cells. Rotenone-induced cell death preceded intracellular accumulation of reactive oxygen species, and antioxidants failed to protect cells, indicating that oxidative stress was minimally involved in rotenone-induced death of SK-N-MC cells. Glycogen synthase kinase 3β (GSK3β), a multifunctional serine/threonine kinase, has been implicated in the pathogenesis of neurodegeneration. We showed that rotenone activated GSK3β by enhancing its phosphorylation at tyrosine 216 while inhibiting phosphorylation at serine 9. Inhibitors of GSK3β and dominant negative (kinase deficient) GSK3β partially protected SK-N-MC cells against rotenone cytotoxicity. Rotenone also induced endoplasmic reticulum (ER) stress which was evident by an increase in phosphorylation of PERK, PKR, and eIF2α as well as the expression of GRP78. Rotenone had a modest effect on the expression of CHOP. An eIF2α siRNA significantly reduced rotenone cytotoxicity. ER stress was experimentally induced by tunicamycin and thapsigargin, but tunicamycin/thapsigargin did not activate GSK3β in SK-N-MC cells. Down-regulation of eIF2α also offered partial protection against rotenone cytotoxicity. Combined treatment of GSK3β inhibitors and eIF2α siRNA provided much greater protection than either treatment alone. Taken together, the results suggest that GSK3β activation and ER stress contribute separately to rotenone cytotoxicity.

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GSK3β and endoplasmic reticulum stress mediate rotenone-induced death of SK-N-MC neuroblastoma cells

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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