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Article: Role of TRPM2 in H2O2-induced cell apoptosis in endothelial cells

TitleRole of TRPM2 in H2O2-induced cell apoptosis in endothelial cells
Authors
KeywordsAnimal cell
Apoptosis
Calcium transport
Cell death
DNA fragmentation
Issue Date2012
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
Citation
PLoS One, 2012, v. 7 n. 8, article no. e43186 How to Cite?
AbstractMelastatin-like transient receptor potential channel 2 (TRPM2) is an oxidant-sensitive and cationic non-selective channel that is expressed in mammalian vascular endothelium. Here we investigated the functional role of TRPM2 channels in hydrogen peroxide (H(2)O(2))-induced cytosolic Ca(2+) ([Ca(2+)](i)) elavation, whole-cell current increase, and apoptotic cell death in murine heart microvessel endothelial cell line H5V. A TRPM2 blocking antibody (TM2E3), which targets the E3 region near the ion permeation pore of TRPM2, was developed. Treatment of H5V cells with TM2E3 reduced the [Ca(2+)](i) rise and whole-cell current change in response to H(2)O(2). Suppressing TRPM2 expression using TRPM2-specific short hairpin RNA (shRNA) had similar inhibitory effect. H(2)O(2)-induced apoptotic cell death in H5V cells was examined using MTT assay, DNA ladder formation analysis, and DAPI-based nuclear DNA condensation assay. Based on these assays, TM2E3 and TRPM2-specific shRNA both showed protective effect against H(2)O(2)-induced apoptotic cell death. TM2E3 and TRPM2-specific shRNA also protect the cells from tumor necrosis factor (TNF)-alpha-induced cell death in MTT assay. In contrast, overexpression of TRPM2 in H5V cells resulted in an increased response in [Ca(2+)](i) and whole-cell currents to H(2)O(2). TRPM2 overexpression also aggravated the H(2)O(2)-induced apoptotic cell death. Downstream pathways following TRPM2 activation was examined. Results showed that TRPM2 activity stimulated caspase-8, caspase-9 and caspase-3. These findings strongly suggest that TRPM2 channel mediates cellular Ca(2+) overload in response to H(2)O(2) and contribute to oxidant-induced apoptotic cell death in vascular endothelial cells. Down-regulating endogenous TRPM2 could be a means to protect the vascular endothelial cells from apoptotic cell death.
Persistent Identifierhttp://hdl.handle.net/10722/169248
ISSN
2015 Impact Factor: 3.057
2015 SCImago Journal Rankings: 1.395
PubMed Central ID
ISI Accession Number ID
Errata

 

DC FieldValueLanguage
dc.contributor.authorSun, Len_US
dc.contributor.authorYau, HY-
dc.contributor.authorWong, MY-
dc.contributor.authorLi, RA-
dc.contributor.authorHuang, Y-
dc.contributor.authorYao, X-
dc.date.accessioned2012-10-18T08:47:07Z-
dc.date.available2012-10-18T08:47:07Z-
dc.date.issued2012en_US
dc.identifier.citationPLoS One, 2012, v. 7 n. 8, article no. e43186en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://hdl.handle.net/10722/169248-
dc.description.abstractMelastatin-like transient receptor potential channel 2 (TRPM2) is an oxidant-sensitive and cationic non-selective channel that is expressed in mammalian vascular endothelium. Here we investigated the functional role of TRPM2 channels in hydrogen peroxide (H(2)O(2))-induced cytosolic Ca(2+) ([Ca(2+)](i)) elavation, whole-cell current increase, and apoptotic cell death in murine heart microvessel endothelial cell line H5V. A TRPM2 blocking antibody (TM2E3), which targets the E3 region near the ion permeation pore of TRPM2, was developed. Treatment of H5V cells with TM2E3 reduced the [Ca(2+)](i) rise and whole-cell current change in response to H(2)O(2). Suppressing TRPM2 expression using TRPM2-specific short hairpin RNA (shRNA) had similar inhibitory effect. H(2)O(2)-induced apoptotic cell death in H5V cells was examined using MTT assay, DNA ladder formation analysis, and DAPI-based nuclear DNA condensation assay. Based on these assays, TM2E3 and TRPM2-specific shRNA both showed protective effect against H(2)O(2)-induced apoptotic cell death. TM2E3 and TRPM2-specific shRNA also protect the cells from tumor necrosis factor (TNF)-alpha-induced cell death in MTT assay. In contrast, overexpression of TRPM2 in H5V cells resulted in an increased response in [Ca(2+)](i) and whole-cell currents to H(2)O(2). TRPM2 overexpression also aggravated the H(2)O(2)-induced apoptotic cell death. Downstream pathways following TRPM2 activation was examined. Results showed that TRPM2 activity stimulated caspase-8, caspase-9 and caspase-3. These findings strongly suggest that TRPM2 channel mediates cellular Ca(2+) overload in response to H(2)O(2) and contribute to oxidant-induced apoptotic cell death in vascular endothelial cells. Down-regulating endogenous TRPM2 could be a means to protect the vascular endothelial cells from apoptotic cell death.-
dc.languageengen_US
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.actionen_US
dc.relation.ispartofPLoS Oneen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subjectAnimal cell-
dc.subjectApoptosis-
dc.subjectCalcium transport-
dc.subjectCell death-
dc.subjectDNA fragmentation-
dc.titleRole of TRPM2 in H2O2-induced cell apoptosis in endothelial cellsen_US
dc.typeArticleen_US
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1932-6203&volume=8&issue=8&spage=e43186&epage=&date=2012&atitle=Role+of+TRPM2+in+H(2)O(2)-Induced+Cell+Apoptosis+in+Endothelial+Cellsen_US
dc.identifier.emailLi, RA: ronaldli@hkucc.hku.hken_US
dc.identifier.emailYao, X: yao2068@cuhk.edu.hk-
dc.identifier.authorityLi, RA=rp01352en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1371/journal.pone.0043186-
dc.identifier.pmid22916222-
dc.identifier.pmcidPMC3423428-
dc.identifier.scopuseid_2-s2.0-84865165313-
dc.identifier.hkuros212192en_US
dc.identifier.volume7en_US
dc.identifier.issue8, article no. e43186en_US
dc.identifier.isiWOS:000307733800052-
dc.publisher.placeUnited States-
dc.relation.erratumdoi:10.1371/annotation/b41e03f8-e0b8-4bf5-b36e-e0fee6364085-

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