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[Role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion-induced apoptosis of human neuroblastoma SH-SY5Y cells].
Nan Fang Yi Ke Da Xue Xue Bao. 2013 Apr; 33(4):479-85.NF

Abstract

OBJECTIVE

To investigate the role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion (MPP⁺)-induced apoptosis of human neuroblastoma SH-SY5Y cells.

METHODS

The viability of SH-SY5Y cells exposed to varying concentrations of MPP⁺ was assessed using MTT colorimetric assay, and MPP⁺-induced cell apoptosis was detected with hoechst 33342 staining and Annexin V+PI assay. Western blotting and rhodamine 123 staining were employed to examine the changes in cellular poly(ADP-ribose) polymerase (PARP) protein expression and mitochondrial membrane potential in response to MPP⁺ exposure. The effects of ruthenium red and/or MPP⁺ on calcium concentration in the cytoplasm, mitochondria and endoplasmic reticulum were evaluated using confocal microscopy.

RESULTS

MPP⁺ induced apoptosis and caused reduced cell viability and mitochondrial membrane potential in SH-SY5Y cells. The cells exposed to MPP⁺ showed a lowered calcium concentration in the cytoplasm and endoplasmic reticulum and an increased mitochondrial Ca²⁺ uptake. Ruthenium red rescued MPP⁺-induced apoptosis and mitochondrial membrane potential reduction, reduced PARP cleavage, and inhibited the increase of mitochondrial matrix free Ca²⁺ in the cells exposed to MPP⁺.

CONCLUSION

Mitochondrial calcium overload plays an important role in MPP⁺-induced apoptosis of SH-SY5Y cells, which is closely associated with dysregulation of intracellular Ca²⁺ homeostasis.

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Authors+Show Affiliations

Xu L
Department of Occupational Health and Medicine, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. xuleizh@126.com
Li W
No affiliation info available
Lin D
No affiliation info available
Zhang H
No affiliation info available
Zou F
No affiliation info available

MeSH

1-Methyl-4-phenylpyridiniumApoptosisCalciumCell Line, TumorCell SurvivalCytoplasmEndoplasmic ReticulumHomeostasisHumansMembrane Potential, MitochondrialPoly(ADP-ribose) Polymerases

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

23644103

Citation

Xu, Lei, et al. "[Role of Calcium Dyshomeostasis in 1-methyl-4-phenylpyridinium Ion-induced Apoptosis of Human Neuroblastoma SH-SY5Y Cells]." Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University, vol. 33, no. 4, 2013, pp. 479-85.
Xu L, Li W, Lin D, et al. [Role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion-induced apoptosis of human neuroblastoma SH-SY5Y cells]. Nan Fang Yi Ke Da Xue Xue Bao. 2013;33(4):479-85.
Xu, L., Li, W., Lin, D., Zhang, H., & Zou, F. (2013). [Role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion-induced apoptosis of human neuroblastoma SH-SY5Y cells]. Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University, 33(4), 479-85.
Xu L, et al. [Role of Calcium Dyshomeostasis in 1-methyl-4-phenylpyridinium Ion-induced Apoptosis of Human Neuroblastoma SH-SY5Y Cells]. Nan Fang Yi Ke Da Xue Xue Bao. 2013;33(4):479-85. PubMed PMID: 23644103.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion-induced apoptosis of human neuroblastoma SH-SY5Y cells]. AU - Xu,Lei, AU - Li,Wenjun, AU - Lin,Danmiao, AU - Zhang,Hongmei, AU - Zou,Fei, PY - 2013/5/7/entrez PY - 2013/5/7/pubmed PY - 2014/7/18/medline SP - 479 EP - 85 JF - Nan fang yi ke da xue xue bao = Journal of Southern Medical University JO - Nan Fang Yi Ke Da Xue Xue Bao VL - 33 IS - 4 N2 - OBJECTIVE: To investigate the role of calcium dyshomeostasis in 1-methyl-4-phenylpyridinium ion (MPP⁺)-induced apoptosis of human neuroblastoma SH-SY5Y cells. METHODS: The viability of SH-SY5Y cells exposed to varying concentrations of MPP⁺ was assessed using MTT colorimetric assay, and MPP⁺-induced cell apoptosis was detected with hoechst 33342 staining and Annexin V+PI assay. Western blotting and rhodamine 123 staining were employed to examine the changes in cellular poly(ADP-ribose) polymerase (PARP) protein expression and mitochondrial membrane potential in response to MPP⁺ exposure. The effects of ruthenium red and/or MPP⁺ on calcium concentration in the cytoplasm, mitochondria and endoplasmic reticulum were evaluated using confocal microscopy. RESULTS: MPP⁺ induced apoptosis and caused reduced cell viability and mitochondrial membrane potential in SH-SY5Y cells. The cells exposed to MPP⁺ showed a lowered calcium concentration in the cytoplasm and endoplasmic reticulum and an increased mitochondrial Ca²⁺ uptake. Ruthenium red rescued MPP⁺-induced apoptosis and mitochondrial membrane potential reduction, reduced PARP cleavage, and inhibited the increase of mitochondrial matrix free Ca²⁺ in the cells exposed to MPP⁺. CONCLUSION: Mitochondrial calcium overload plays an important role in MPP⁺-induced apoptosis of SH-SY5Y cells, which is closely associated with dysregulation of intracellular Ca²⁺ homeostasis. SN - 1673-4254 UR - https://www.unboundmedicine.com/medline/citation/23644103/[Role_of_calcium_dyshomeostasis_in_1_methyl_4_phenylpyridinium_ion_induced_apoptosis_of_human_neuroblastoma_SH_SY5Y_cells]_ L2 - http://www.diseaseinfosearch.org/result/8994 DB - PRIME DP - Unbound Medicine ER -