Human vascular smooth muscle cells express an estrogen receptor isoform

In women, estrogen (E2) exerts a clinically relevant anti-atherogenic effect. The atheroprotective effects of E2 are mediated both by E2-induced changes in systemic factors and by direct effects of E2 on the blood vessel wall. In studies to characterize E2 signaling pathways in vascular smooth muscle cells (VSMC), we recently demonstrated that human VSMC express a functional estrogen receptor [1]. In the present study, we applied a reverse transcription/PCR-based strategy to identify isoforms of the E2 receptor in human VSMC. We now report that in addition to the classical E2 receptor, human VSMC derived from both mammary artery and saphenous vein express an estrogen receptor isoform containing an in-frame deletion of Exon 4 (ER delta 4). RNase protection assays confirm the presence of ER delta 4 message in VSMC and demonstrate it is nearly as abundant as the classical E2 receptor. Transient transfection experiments in VSMC and HeLa cells demonstrate that, in contrast to the classical 67 kDa nuclear-localized E2 receptor, ER delta 4: (a) is a 55 kDa protein that is widely distributed throughout the cell; (b) does not transactivate an E2 response element-driven reporter plasmid in response to E2; and (c) does not modulate transactivation of the ERE-reporter by the classical (wild type) estrogen receptor. Thus, human VSMC express an E2 receptor isoform that does not appear to alter gene transcription. The presence of a novel isoform of the E2 receptor may have important implications for studies of E2-mediated signaling in VSMC.