Orimo A, Inoue S, Ikegami A, Hosoi T, Akishita M, Ouchi Y, Muramatsu M, Orimo H
Department of Geriatrics, Faculty of Medicine, University of Tokyo, Japan.
Biochem Biophys Res Commun. 1993 Sep 15;195(2):730-6. doi: 10.1006/bbrc.1993.2106.
ER mRNA was detected as 6.0 kb band by Northern blot analysis in vascular smooth muscle cells (VSMC) derived from rat aorta. The presence of ER mRNA in VSMC was confirmed by reverse transcriptase-polymerase chain reaction using specific primers for rat ER cDNA. In addition, the immunocytochemistry of ER was performed in VSMC using a monoclonal anti-ER antibody which recognizes DNA-binding domain of ER. The immunoreactivity was distributed in the cytoplasm as well as in the nuclei. Thus, the expression of ER in VSMC was demonstrated at both the protein and the mRNA level. Furthermore, the expression of c-fos mRNA in VSMC was found up-regulated by 17 beta-estradiol treatment within 30 min. The observation that VSMC possess ER and respond to estrogen supports the idea that estrogen may directly influence vascular cell system through the ER.
通过Northern印迹分析,在源自大鼠主动脉的血管平滑肌细胞(VSMC)中检测到ER mRNA为6.0 kb条带。使用针对大鼠ER cDNA的特异性引物,通过逆转录聚合酶链反应证实了VSMC中存在ER mRNA。此外,使用识别ER DNA结合域的单克隆抗ER抗体在VSMC中进行ER的免疫细胞化学分析。免疫反应性分布在细胞质和细胞核中。因此,在蛋白质和mRNA水平均证实了VSMC中ER的表达。此外,发现17β-雌二醇处理30分钟内VSMC中c-fos mRNA的表达上调。VSMC具有ER并对雌激素作出反应这一观察结果支持了雌激素可能通过ER直接影响血管细胞系统的观点。