Oviedo Pilar J, Hermenegildo Carlos, Tarín Juan J, Cano Antonio
Department of Pediatrics, Obstetrics and Gynecology, Faculty of Medicine, University of Valencia, Valencia, Spain.
Fertil Steril. 2007 Aug;88(2):326-32. doi: 10.1016/j.fertnstert.2006.11.185. Epub 2007 Apr 18.
To examine the proliferative effect of of raloxifene on human umbilical-vein endothelial cells (HUVECs), and to investigate whether there is an associated increased expression of some key regulators of the cell cycle.
Cell culture for different incubation times.
University research laboratory.
PATIENT(S): Sources of HUVECs.
INTERVENTION(S): Measurement of cell proliferation, of protein levels of cyclin A, cyclin B1, cyclin D1, cyclin-dependent protein kinase (CDK) 2, CDK4, and p27(Kip1), and of messenger RNA expression of cyclin A, cyclin B1, and p27(Kip1).
MAIN OUTCOME MEASURE(S): Cell proliferation was measured by the 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the bromo-2'-deoxyuridine assay, and the sodium 3'-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis (4-methoxy-6-nitro) benzene sulfonic acid hydrate assay. Changes in protein expression were measured by immunoblotting, and modifications in gene expression were measured by quantitative real-time polymerase chain reaction.
RESULT(S): Both 1 nM and 10 nM of either E(2) or raloxifene achieved a similar increase in cell proliferation. The pure antiestrogen ICI 182780 only blocked the E(2)-induced proliferative effect. Western blot experiments detected an increase in the expression of only cyclin A and B1, and a decreasing trend for p27(Kip1). Enhancements in gene expression were observed in response to E(2) and raloxifene for cyclin A and B1. No significant changes were found for p27(Kip1). The ICI 182780 effectively abrogated the increased gene expression associated with E(2) for cyclin B1, but not for cyclin A. In contrast, ICI 182780 was ineffective in the case of raloxifene.
CONCLUSION(S): Raloxifene increased the proliferation of HUVECs in association with enhanced gene expression of cyclins A and B1.
研究雷洛昔芬对人脐静脉内皮细胞（HUVECs）的增殖作用，并探讨细胞周期某些关键调节因子的表达是否相应增加。
不同孵育时间的细胞培养。
大学研究实验室。
HUVECs来源。
测量细胞增殖、细胞周期蛋白A、细胞周期蛋白B1、细胞周期蛋白D1、细胞周期蛋白依赖性蛋白激酶（CDK）2、CDK4和p27（Kip1）的蛋白水平，以及细胞周期蛋白A、细胞周期蛋白B1和p27（Kip1）的信使核糖核酸表达。
通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐试验、溴脱氧尿苷试验和3'-[1-（苯氨基羰基）-3,4-四氮唑]-双（4-甲氧基-6-硝基）苯磺酸水合物试验测量细胞增殖。通过免疫印迹法测量蛋白表达变化，通过定量实时聚合酶链反应测量基因表达变化。
1 nM和10 nM的雌二醇（E2）或雷洛昔芬均使细胞增殖有相似程度的增加。纯抗雌激素ICI 182780仅阻断E2诱导的增殖作用。蛋白质印迹实验检测到仅细胞周期蛋白A和B1的表达增加，p27（Kip1）有下降趋势。观察到E2和雷洛昔芬作用后细胞周期蛋白A和B1的基因表达增强。p27（Kip1）未发现显著变化。ICI 182780有效消除了与E2相关的细胞周期蛋白B1基因表达增加，但对细胞周期蛋白A无效。相比之下，ICI 182780对雷洛昔芬无效。
雷洛昔芬增加HUVECs的增殖，同时细胞周期蛋白A和B1的基因表达增强。
