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Article: Endothelin-1 pathway in human alveolar epithelial cell line A549 and human umbilical vein endothelial cells

TitleEndothelin-1 pathway in human alveolar epithelial cell line A549 and human umbilical vein endothelial cells
Authors
Issue Date2000
Citation
Acta Pharmacologica Sinica, 2000, v. 21 n. 6, p. 499-506 How to Cite?
AbstractAIM: This study was designed to characterize the endothelin pathway in an immortalized human adenocarcinoma-derived alveolar epithelial cell line (A549) and human umbilical vein endothelial cell line (HUVEC). METHODS: The release of ET-1 and big-ET-1 was measured in the incubation medium of both cell lines. The expression of mRNAs coding for the endothelin isoforms (hppET-1, -2, -3), the endothelin converting enzymes (hECE-la, b, c, and d) and the hET(A) and hET(B) receptors was investigated using RT-PCR. The expression of ECE-1 mRNA in various human tissues and in A549 cells was investigated by Northern blot analysis and the subcellular localization of ECE-1 in A549 cells was investigated by immunoblotting using a polyclonal antibody. RESULTS: Under control conditions, HUVEC release both ET-1 and big- ET-1 (ratio 5 to 1) while in A549 cells the big-ET-1 levels were below the threshold of detection. The release of these two peptides was minimally affected by various inhibitors of peptidases. However, in both cell lines phosphoramidon produced a concentration-dependent inhibition of ET-1 release and an enhanced accumulation of big-ET-1. Both HUVEC and A549 cells express the mRNAs for ppET-1, ET-A, and ET-B receptor subtypes and ECE-1 (isoforms ECE-1b, c and/or d). In addition, in HUVEC the mRNAs for ppET-2 and for the isoform ECE-1a were also detected. In A549 cells, ECE-1 had a preferential subcellular localization in the membrane fraction but was not detected in the cytosol. CONCLUSION: Both A549 and HUVEC produce and release endothelin-1 through a specific enzymatic pathway, whether or not ECE-1 is the only enzyme involved remains to be determined. A549 might be used as a screening assay for drug discovery such as for inhibitors of endothelin-1 release.
Persistent Identifierhttp://hdl.handle.net/10722/171241
ISSN
2000 Impact Factor: 0.485
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorDeprezRoy, Ien_US
dc.contributor.authorCoge, Fen_US
dc.contributor.authorBertry, Len_US
dc.contributor.authorGalizzi, JPen_US
dc.contributor.authorFeletou, Men_US
dc.contributor.authorVanhoutte, PMen_US
dc.contributor.authorCanet, Een_US
dc.date.accessioned2012-10-30T06:12:54Z-
dc.date.available2012-10-30T06:12:54Z-
dc.date.issued2000en_US
dc.identifier.citationActa Pharmacologica Sinica, 2000, v. 21 n. 6, p. 499-506en_US
dc.identifier.issn0253-9756en_US
dc.identifier.urihttp://hdl.handle.net/10722/171241-
dc.description.abstractAIM: This study was designed to characterize the endothelin pathway in an immortalized human adenocarcinoma-derived alveolar epithelial cell line (A549) and human umbilical vein endothelial cell line (HUVEC). METHODS: The release of ET-1 and big-ET-1 was measured in the incubation medium of both cell lines. The expression of mRNAs coding for the endothelin isoforms (hppET-1, -2, -3), the endothelin converting enzymes (hECE-la, b, c, and d) and the hET(A) and hET(B) receptors was investigated using RT-PCR. The expression of ECE-1 mRNA in various human tissues and in A549 cells was investigated by Northern blot analysis and the subcellular localization of ECE-1 in A549 cells was investigated by immunoblotting using a polyclonal antibody. RESULTS: Under control conditions, HUVEC release both ET-1 and big- ET-1 (ratio 5 to 1) while in A549 cells the big-ET-1 levels were below the threshold of detection. The release of these two peptides was minimally affected by various inhibitors of peptidases. However, in both cell lines phosphoramidon produced a concentration-dependent inhibition of ET-1 release and an enhanced accumulation of big-ET-1. Both HUVEC and A549 cells express the mRNAs for ppET-1, ET-A, and ET-B receptor subtypes and ECE-1 (isoforms ECE-1b, c and/or d). In addition, in HUVEC the mRNAs for ppET-2 and for the isoform ECE-1a were also detected. In A549 cells, ECE-1 had a preferential subcellular localization in the membrane fraction but was not detected in the cytosol. CONCLUSION: Both A549 and HUVEC produce and release endothelin-1 through a specific enzymatic pathway, whether or not ECE-1 is the only enzyme involved remains to be determined. A549 might be used as a screening assay for drug discovery such as for inhibitors of endothelin-1 release.en_US
dc.languageengen_US
dc.relation.ispartofActa Pharmacologica Sinicaen_US
dc.subject.meshAdenocarcinoma - Pathologyen_US
dc.subject.meshAspartic Acid Endopeptidases - Biosynthesis - Geneticsen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshEndothelin-1 - Secretionen_US
dc.subject.meshEndothelins - Biosynthesis - Geneticsen_US
dc.subject.meshEndothelium, Vascular - Cytology - Metabolismen_US
dc.subject.meshGlycopeptides - Pharmacologyen_US
dc.subject.meshHumansen_US
dc.subject.meshLung Neoplasms - Pathologyen_US
dc.subject.meshMetalloendopeptidasesen_US
dc.subject.meshProtein Precursors - Biosynthesis - Geneticsen_US
dc.subject.meshRna, Messenger - Biosynthesisen_US
dc.subject.meshTumor Cells, Cultureden_US
dc.subject.meshUmbilical Veins - Cytology - Metabolismen_US
dc.titleEndothelin-1 pathway in human alveolar epithelial cell line A549 and human umbilical vein endothelial cellsen_US
dc.typeArticleen_US
dc.identifier.emailVanhoutte, PM:vanhoutt@hku.hken_US
dc.identifier.authorityVanhoutte, PM=rp00238en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.pmid11360683-
dc.identifier.scopuseid_2-s2.0-0034045779en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0034045779&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume21en_US
dc.identifier.issue6en_US
dc.identifier.spage499en_US
dc.identifier.epage506en_US
dc.identifier.isiWOS:000087561300003-
dc.identifier.scopusauthoridDeprezRoy, I=6504471806en_US
dc.identifier.scopusauthoridCoge, F=6602987151en_US
dc.identifier.scopusauthoridBertry, L=6504401404en_US
dc.identifier.scopusauthoridGalizzi, JP=7004145226en_US
dc.identifier.scopusauthoridFeletou, M=7006461826en_US
dc.identifier.scopusauthoridVanhoutte, PM=7202304247en_US
dc.identifier.scopusauthoridCanet, E=7006072145en_US

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