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Article: Role of TRPM2 in H2O2-induced cell apoptosis in endothelial cells
Title | Role of TRPM2 in H2O2-induced cell apoptosis in endothelial cells |
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Authors | |
Keywords | Animal cell Apoptosis Calcium transport Cell death DNA fragmentation |
Issue Date | 2012 |
Publisher | Public 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? |
Abstract | Melastatin-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 Identifier | http://hdl.handle.net/10722/169248 |
ISSN | 2023 Impact Factor: 2.9 2023 SCImago Journal Rankings: 0.839 |
PubMed Central ID | |
ISI Accession Number ID | |
Errata |
DC Field | Value | Language |
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dc.contributor.author | Sun, L | en_US |
dc.contributor.author | Yau, HY | - |
dc.contributor.author | Wong, MY | - |
dc.contributor.author | Li, RA | - |
dc.contributor.author | Huang, Y | - |
dc.contributor.author | Yao, X | - |
dc.date.accessioned | 2012-10-18T08:47:07Z | - |
dc.date.available | 2012-10-18T08:47:07Z | - |
dc.date.issued | 2012 | en_US |
dc.identifier.citation | PLoS One, 2012, v. 7 n. 8, article no. e43186 | en_US |
dc.identifier.issn | 1932-6203 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/169248 | - |
dc.description.abstract | Melastatin-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.language | eng | en_US |
dc.publisher | Public Library of Science. The Journal's web site is located at http://www.plosone.org/home.action | en_US |
dc.relation.ispartof | PLoS ONE | en_US |
dc.rights | This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. | - |
dc.subject | Animal cell | - |
dc.subject | Apoptosis | - |
dc.subject | Calcium transport | - |
dc.subject | Cell death | - |
dc.subject | DNA fragmentation | - |
dc.title | Role of TRPM2 in H2O2-induced cell apoptosis in endothelial cells | en_US |
dc.type | Article | en_US |
dc.identifier.openurl | http://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+Cells | en_US |
dc.identifier.email | Li, RA: ronaldli@hkucc.hku.hk | en_US |
dc.identifier.email | Yao, X: yao2068@cuhk.edu.hk | - |
dc.identifier.authority | Li, RA=rp01352 | en_US |
dc.description.nature | published_or_final_version | - |
dc.identifier.doi | 10.1371/journal.pone.0043186 | - |
dc.identifier.pmid | 22916222 | - |
dc.identifier.pmcid | PMC3423428 | - |
dc.identifier.scopus | eid_2-s2.0-84865165313 | - |
dc.identifier.hkuros | 212192 | en_US |
dc.identifier.volume | 7 | en_US |
dc.identifier.issue | 8, article no. e43186 | en_US |
dc.identifier.isi | WOS:000307733800052 | - |
dc.publisher.place | United States | - |
dc.relation.erratum | doi:10.1371/annotation/b41e03f8-e0b8-4bf5-b36e-e0fee6364085 | - |
dc.identifier.issnl | 1932-6203 | - |