The G protein-coupled receptor GPR30 inhibits proliferation of estrogen receptor-positive breast cancer cells.
The G protein-coupled receptor GPR30 inhibits proliferation of estrogen receptor-positive breast cancer cells.
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DOI：
10.1158/0008-5472.can-09-3068
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发表时间：
2010-02-01
期刊：
影响因子：
11.2
通讯作者：
Jordan VC
中科院分区：
文献类型：
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作者：
Ariazi EA;Brailoiu E;Yerrum S;Shupp HA;Slifker MJ;Cunliffe HE;Black MA;Donato AL;Arterburn JB;Oprea TI;Prossnitz ER;Dun NJ;Jordan VC
G蛋白偶联受体GPR30（GPER）可与17β - 雌二醇（E2）结合，但不同于经典的雌激素受体（ERα和ERβ）。GPR30能够介导E2诱导的非基因组信号传导，但其在ERα阳性乳腺癌中的作用仍不明确。来自5个队列共1250例乳腺癌的基因表达微阵列数据显示，GPR30表达增加与ERα阳性状态之间存在关联。因此，我们利用G - 1和己烯雌酚（分别选择性激活GPR30和ER的配体）以及小干扰RNA（siRNAs），在ER阳性的MCF - 7乳腺癌细胞中研究了GPR30在雌激素活性中的作用。在表达研究中，E2和己烯雌酚而非G - 1可下调ER和GPR30，表明这是由ER介导的。在Ca2 +动员研究中，GPR30而非ERα介导E2诱导的Ca2 +反应，因为E2、4 - 羟基他莫昔芬（也激活GPR30）和G - 1，但不是己烯雌酚，不仅在MCF - 7细胞中，而且在ER阴性的SKBr3细胞中都能引起胞质Ca2 +增加。此外，在MCF - 7细胞中，GPR30缺失阻断了E2和G - 1诱导的Ca2 +动员，但ERα缺失则没有。有趣的是，在ER阳性的MCF - 7细胞中，GPR30偶联的Ca2 +反应是持续的且由肌醇三磷酸受体介导，而在ER阴性的SKBr3细胞中则是短暂的且由兰尼碱受体介导。涉及GPR30缺失的增殖研究表明，GPR30的作用是促进SKBr3细胞生长，但抑制MCF - 7细胞生长。与此相符的是，G - 1可能通过诱导p53和p21显著抑制MCF - 7细胞生长。此外，流式细胞术显示G - 1在G1期阻断了MCF - 7细胞周期进程。因此，GPR30拮抗ERα阳性乳腺癌的生长，可能代表了一种对抗这种疾病的新靶点。
The G protein-coupled receptor GPR30 (GPER) binds 17β-estradiol (E2), yet differs from classical estrogen receptors (ERα and ERβ). GPR30 can mediate E2-induced non-genomic signaling, but its role in ERα-positive breast cancer remains unclear. Gene expression microarray data from 5 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, ligands which selectively activate GPR30 and ER, respectively, and small interfering RNAs (siRNAs). In expression studies, E2 and diethylstilbestrol but not G-1 down-regulated both ER and GPR30, indicating this was ER mediated. In Ca2+ mobilization studies, GPR30 but not ERα mediated E2-induced Ca2+ responses, since E2, 4-hydroxytamoxifen (also 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- and G-1-induced Ca2+ 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 GPR30's role 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.