TY - JOUR
T1 - The G protein-coupled receptor GPR30 inhibits proliferation of estrogen receptor-positive breast cancer cells
AU - Ariazi, Eric A.
AU - Brailoiu, Eugen
AU - Yerrum, Smitha
AU - Shupp, Heather A.
AU - Slifker, Michael J.
AU - Cunliffe, Heather E.
AU - Black, Michael A.
AU - Donato, Anne L.
AU - Arterburn, Jeffrey B.
AU - Oprea, Tudor I.
AU - Prossnitz, Eric R.
AU - Dun, Nae J.
AU - Jordan, V. Craig
PY - 2010/2/1
Y1 - 2010/2/1
N2 - The G protein-coupled receptor GPR30 binds 17β-estradiol (E 2) yet differs from classic estrogen receptors (ERα and ERβ). GPR30 can mediate E2-induced nongenomic signaling, but its role in ERα-positive breast cancer remains unclear. Gene expression microarray data from five cohorts comprising 1,250 breast carcinomas showed an association between increased GPR30 expression and ERα-positive status. We therefore examined GPR30 in estrogenic activities in ER-positive MCF-7 breast cancer cells using G-1 and diethylstilbestrol (DES), ligands that selectively activate GPR30 and ER, respectively, and small interfering RNAs. In expression studies, E2 and DES, but not G-1, transiently downregulated both ER and GPR30, indicating that this was ER mediated. In Ca2+ mobilization studies, GPR30, but not ERα, mediated E2-induced Ca 2+ responses because E2, 4-hydroxytamoxifen (activates GPR30), and G-1, but not DES, elicited cytosolic Ca2+ increases not only in MCF-7 cells but also in ER-negative SKBr3 cells. Additionally, in MCF-7 cells, GPR30 depletion blocked E2-induced and G-1-induced C a2+ mobilization, but ERα depletion did not. Interestingly, GPR30-coupled Ca2+ responses were sustained and inositol triphosphate receptor mediated in ER-positive MCF-7 cells but transitory and ryanodine receptor mediated in ER-negative SKBr3 cells. Proliferation studies involving GPR30 depletion indicated that the role of GPR30 was to promote SKBr3 cell growth but reduce MCF-7 cell growth. Supporting this, G-1 profoundly inhibited MCF-7 cell growth, potentially via p53 and p21 induction. Further, flow cytometry showed that G-1 blocked MCF-7 cell cycle progression at the G 1 phase. Thus, GPR30 antagonizes growth of ERα-positive breast cancer and may represent a new target to combat this disease.
AB - The G protein-coupled receptor GPR30 binds 17β-estradiol (E 2) yet differs from classic estrogen receptors (ERα and ERβ). GPR30 can mediate E2-induced nongenomic signaling, but its role in ERα-positive breast cancer remains unclear. Gene expression microarray data from five cohorts comprising 1,250 breast carcinomas showed an association between increased GPR30 expression and ERα-positive status. We therefore examined GPR30 in estrogenic activities in ER-positive MCF-7 breast cancer cells using G-1 and diethylstilbestrol (DES), ligands that selectively activate GPR30 and ER, respectively, and small interfering RNAs. In expression studies, E2 and DES, but not G-1, transiently downregulated both ER and GPR30, indicating that this was ER mediated. In Ca2+ mobilization studies, GPR30, but not ERα, mediated E2-induced Ca 2+ responses because E2, 4-hydroxytamoxifen (activates GPR30), and G-1, but not DES, elicited cytosolic Ca2+ increases not only in MCF-7 cells but also in ER-negative SKBr3 cells. Additionally, in MCF-7 cells, GPR30 depletion blocked E2-induced and G-1-induced C a2+ mobilization, but ERα depletion did not. Interestingly, GPR30-coupled Ca2+ responses were sustained and inositol triphosphate receptor mediated in ER-positive MCF-7 cells but transitory and ryanodine receptor mediated in ER-negative SKBr3 cells. Proliferation studies involving GPR30 depletion indicated that the role of GPR30 was to promote SKBr3 cell growth but reduce MCF-7 cell growth. Supporting this, G-1 profoundly inhibited MCF-7 cell growth, potentially via p53 and p21 induction. Further, flow cytometry showed that G-1 blocked MCF-7 cell cycle progression at the G 1 phase. Thus, GPR30 antagonizes growth of ERα-positive breast cancer and may represent a new target to combat this disease.
UR - http://www.scopus.com/inward/record.url?scp=76249115409&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=76249115409&partnerID=8YFLogxK
U2 - 10.1158/0008-5472.CAN-09-3068
DO - 10.1158/0008-5472.CAN-09-3068
M3 - Article
C2 - 20086172
AN - SCOPUS:76249115409
SN - 0008-5472
VL - 70
SP - 1184
EP - 1194
JO - Cancer Research
JF - Cancer Research
IS - 3
ER -