E4F1, a novel estrogen-responsive gene in possible atheroprotection, revealed by microarray analysis.
E4F1, a novel estrogen-responsive gene in possible atheroprotection, revealed by microarray analysis.
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DOI：
10.1016/s0002-9440(10)63253-1
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发表时间：
2004-12
期刊：
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通讯作者：
Yasuhiro Nakamura;K. Igarashi;Takashi Suzuki;J. Kanno;Tohru Inoue;C. Tazawa;M. Saruta;T. Ando;N. Moriyama;T. Furukawa;M. Ono;T. Moriya;K. Ito;H. Saito;T. Ishibashi;Shoki Takahashi;S. Yamada;H. Sasano
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文献类型：
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作者：
Yasuhiro Nakamura;K. Igarashi;Takashi Suzuki;J. Kanno;Tohru Inoue;C. Tazawa;M. Saruta;T. Ando;N. Moriyama;T. Furukawa;M. Ono;T. Moriya;K. Ito;H. Saito;T. Ishibashi;Shoki Takahashi;S. Yamada;H. Sasano
雌激素主要通过雌激素受体（ER）参与抑制血管平滑肌细胞（VSMC）增殖，但其具体机制尚不清楚。因此，本研究采用基因芯片技术，对雌激素处理的ERα和ERβ阳性的两种培养的人VSMCs进行雌激素反应基因的检测。我们还使用定量逆转录-聚合酶链反应（RT-PCR）来评估选择性靶基因（TG）在这些细胞中的mRNA水平。我们进一步研究了TG产物是否参与使用TG转染的小干扰RNA（siRNA）抑制增殖。我们随后使用定量RT-PCR和原位杂交分析，以评估这些基因产物在人主动脉的表达。基因芯片和RT-PCR分析显示，E4 F1仅在ERα阳性的VSMCs中表达显著增加。使用siRNA阻断E4 F1抑制雌激素对ERα阳性VSMC增殖的抑制。绝经前女性主动脉E4 F1 mRNA表达丰富，动脉粥样硬化程度较轻。因此，E4 F1被认为是雌激素反应基因之一，参与ERα介导的VSMC增殖抑制，并可能在雌激素相关的动脉粥样硬化保护中发挥重要作用。
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 ERα, and the other for ERβ, 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 ERα-positive VSMCs by estrogens in both microarray and RT-PCR analyses. Blocking of E4F1 using siRNA suppressed estrogenic inhibition of ERα-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 ERα-mediated inhibition of VSMC proliferation and may play an important role in estrogen-related atheroprotection of human aorta.