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(American Journal of Pathology. 2004;165:2019-2031.)
© 2004 American Society for Investigative Pathology

E4F1, a Novel Estrogen-Responsive Gene in Possible Atheroprotection, Revealed by Microarray Analysis

Yasuhiro Nakamura*{dagger}, Katsuhide Igarashi{ddagger}, Takashi Suzuki*, Jun Kanno{ddagger}, Tohru Inoue§, Chika Tazawa*, Masayuki Saruta*, Tomoko Ando{ddagger}, Noriko Moriyama{ddagger}, Toru Furukawa, Masao Ono*, Takuya Moriya*, Kiyoshi Ito||, Haruo Saito{dagger}, Tadashi Ishibashi{dagger}, Shoki Takahashi{dagger}, Shogo Yamada{dagger} and Hironobu Sasano*

From the Departments of Pathology,* Radiology,{dagger} Molecular Pathology, and Gynecology,|| Tohoku University School of Medicine, Sendai; and the Division of Toxicology{ddagger} and the Biological Safety Research Center,§ National Institute of Health Sciences, Tokyo, Japan

Estrogen has been postulated to be involved in inhibition of vascular smooth muscle cell (VSMC) proliferation mainly via estrogen receptor (ER), but the detailed mechanism has remained primarily unknown. Therefore, in this study, microarray analysis was used in two types of cultured human VSMCs: one positive for ER{alpha}, and the other for ERß, which were treated by estrogens to detect the estrogen-responsive genes. We also used quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to evaluate mRNA levels of selective target gene (TG) in these cells. We further studied whether the TG product was involved in inhibition of proliferation using small interfering RNA (siRNA) of the TG transfection. We subsequently used quantitative RT-PCR and in situ hybridization analysis to evaluate the expression of these gene products in human aorta. E4F1, a possible inducer of cell growth arrest, was markedly increased only in ER{alpha}-positive VSMCs by estrogens in both microarray and RT-PCR analyses. Blocking of E4F1 using siRNA suppressed estrogenic inhibition of ER{alpha}-positive VSMC proliferation. E4F1 mRNA was abundant in premenopausal female aorta with mild atherosclerotic changes. E4F1 is therefore considered one of the estrogen-responsive genes involving ER{alpha}-mediated inhibition of VSMC proliferation and may play an important role in estrogen-related atheroprotection of human aorta.




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