Abstract
OBJECTIVE - : Chronic therapy with synthetic glucocorticoids has been associated with cardiovascular side effects, although differential interindividual susceptibility to glucocorticoids has been observed. The objective of this study was to identify the molecular mechanisms leading to differential glucocorticoid responses in endothelial cells. APPROACH AND RESULTS - : We tested the sensitivity of 42 human umbilical vein endothelial cells (HUVECs) to dexamethasone as determined by changes in gene expression, promoter transactivation, and procoagulant activity. We identified that 16 HUVECs were sensitive in every test, 14 HUVECs were sensitive in at least 1 test and 12 HUVECs were resistant in every test to dexamethasone. Nuclear translocation assays revealed that Dex-sensitive HUVECs have higher basal and Dex-stimulated levels of nuclear glucocorticoid receptor compared with Dex-resistant HUVECs. Cycloheximide assays revealed that Dex-resistant HUVECs have significantly shorter glucocorticoid receptor protein half-lives than Dex-sensitive HUVECs. Dex-resistant HUVECs have a stronger interaction of glucocorticoid receptor with the proteasomal recruiting protein, BCL2-associated athanogene 1 (BAG1), as shown by immunoprecipitation assays. Silencing BAG1 expression increased Dex-sensitivity in resistant HUVECs, whereas BAG1 overexpression decreased Dex-sensitivity in sensitive HUVECs. Finally, Dex-resistant HUVECs presented higher BAG1 expression than Dex-sensitive HUVECs. CONCLUSIONS - : In vitro endothelial sensitivity to Dex varies within individuals and is inversely proportional to BAG1 protein expression and glucocorticoid receptor protein turnover.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1046-1055 |
| Number of pages | 10 |
| Journal | Arteriosclerosis, Thrombosis, and Vascular Biology |
| Volume | 33 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2013 |
| Externally published | Yes |
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Keywords
- coagulation
- endothelium
- glucocorticoids
- sensitivity
ASJC Scopus subject areas
- Cardiology and Cardiovascular Medicine
Cite this
Role of BCL2-associated athanogene 1 in differential sensitivity of human endothelial cells to glucocorticoids. / Mata-Greenwood, Eugenia; Stewart, John M.; Steinhorn, Robin H; Pearce, William J.
In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 33, No. 5, 05.2013, p. 1046-1055.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Role of BCL2-associated athanogene 1 in differential sensitivity of human endothelial cells to glucocorticoids
AU - Mata-Greenwood, Eugenia
AU - Stewart, John M.
AU - Steinhorn, Robin H
AU - Pearce, William J.
PY - 2013/5
Y1 - 2013/5
N2 - OBJECTIVE - : Chronic therapy with synthetic glucocorticoids has been associated with cardiovascular side effects, although differential interindividual susceptibility to glucocorticoids has been observed. The objective of this study was to identify the molecular mechanisms leading to differential glucocorticoid responses in endothelial cells. APPROACH AND RESULTS - : We tested the sensitivity of 42 human umbilical vein endothelial cells (HUVECs) to dexamethasone as determined by changes in gene expression, promoter transactivation, and procoagulant activity. We identified that 16 HUVECs were sensitive in every test, 14 HUVECs were sensitive in at least 1 test and 12 HUVECs were resistant in every test to dexamethasone. Nuclear translocation assays revealed that Dex-sensitive HUVECs have higher basal and Dex-stimulated levels of nuclear glucocorticoid receptor compared with Dex-resistant HUVECs. Cycloheximide assays revealed that Dex-resistant HUVECs have significantly shorter glucocorticoid receptor protein half-lives than Dex-sensitive HUVECs. Dex-resistant HUVECs have a stronger interaction of glucocorticoid receptor with the proteasomal recruiting protein, BCL2-associated athanogene 1 (BAG1), as shown by immunoprecipitation assays. Silencing BAG1 expression increased Dex-sensitivity in resistant HUVECs, whereas BAG1 overexpression decreased Dex-sensitivity in sensitive HUVECs. Finally, Dex-resistant HUVECs presented higher BAG1 expression than Dex-sensitive HUVECs. CONCLUSIONS - : In vitro endothelial sensitivity to Dex varies within individuals and is inversely proportional to BAG1 protein expression and glucocorticoid receptor protein turnover.
AB - OBJECTIVE - : Chronic therapy with synthetic glucocorticoids has been associated with cardiovascular side effects, although differential interindividual susceptibility to glucocorticoids has been observed. The objective of this study was to identify the molecular mechanisms leading to differential glucocorticoid responses in endothelial cells. APPROACH AND RESULTS - : We tested the sensitivity of 42 human umbilical vein endothelial cells (HUVECs) to dexamethasone as determined by changes in gene expression, promoter transactivation, and procoagulant activity. We identified that 16 HUVECs were sensitive in every test, 14 HUVECs were sensitive in at least 1 test and 12 HUVECs were resistant in every test to dexamethasone. Nuclear translocation assays revealed that Dex-sensitive HUVECs have higher basal and Dex-stimulated levels of nuclear glucocorticoid receptor compared with Dex-resistant HUVECs. Cycloheximide assays revealed that Dex-resistant HUVECs have significantly shorter glucocorticoid receptor protein half-lives than Dex-sensitive HUVECs. Dex-resistant HUVECs have a stronger interaction of glucocorticoid receptor with the proteasomal recruiting protein, BCL2-associated athanogene 1 (BAG1), as shown by immunoprecipitation assays. Silencing BAG1 expression increased Dex-sensitivity in resistant HUVECs, whereas BAG1 overexpression decreased Dex-sensitivity in sensitive HUVECs. Finally, Dex-resistant HUVECs presented higher BAG1 expression than Dex-sensitive HUVECs. CONCLUSIONS - : In vitro endothelial sensitivity to Dex varies within individuals and is inversely proportional to BAG1 protein expression and glucocorticoid receptor protein turnover.
KW - coagulation
KW - endothelium
KW - glucocorticoids
KW - sensitivity
UR - http://www.scopus.com/inward/record.url?scp=84876285422&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84876285422&partnerID=8YFLogxK
U2 - 10.1161/ATVBAHA.113.301247
DO - 10.1161/ATVBAHA.113.301247
M3 - Article
C2 - 23493285
AN - SCOPUS:84876285422
VL - 33
SP - 1046
EP - 1055
JO - Arteriosclerosis, Thrombosis, and Vascular Biology
JF - Arteriosclerosis, Thrombosis, and Vascular Biology
SN - 1079-5642
IS - 5
ER -