Recombinant baculoviruses used to study estrogen receptor function in human osteosarcoma cells
Recombinant baculoviruses used to study estrogen receptor function in human osteosarcoma cells
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DOI：
10.1089/154065803772613435
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发表时间：
2003-12-01
影响因子：
1.8
通讯作者：
Lorenz, JJ
中科院分区：
文献类型：
--
作者：
Clay, WC;Condreay, JP;Lorenz, JJ
我们报告，修改杆状病毒，称为BacMam病毒，可以有效地将多个基因到哺乳动物细胞中产生异源转录因子/报告基因系统。使用人雌激素受体（ER）作为模型核受体，我们证明了这种方法可以成功地应用于检测开发Saos-2人骨肉瘤细胞。构建了含有人ER亚型ER α和ER β的全长cDNA的BacMam病毒，以及含有ER应答报告基因盒的第三种BacMam病毒。使用这些病毒，我们发现BacMam-ER表达/报告基因构建体可用于分析激动剂17 β-雌二醇和部分激动剂雷洛昔芬在人Saos-2细胞中的作用。使用基于BacMam的系统获得的测定数据与使用标准DNA转染获得的测定数据的比较表明，这两个系统在功能上是等效的，分别给出了雌激素和雌激素加雷洛昔芬治疗的相当的EC 50和IC 50值。我们的研究结果表明，BacMam介导的基因转移提供了一种新的和有效的方法，用于交付核受体和相关基因的哺乳动物细胞为基础的检测开发。
We report that modified baculoviruses, termed BacMam viruses, can efficiently deliver multiple genes into mammalian cells to generate a heterologous transcription factor/reporter gene system. Using human estrogen receptor (ER) as a model nuclear receptor, we demonstrate how this approach can be successfully applied to assay development in Saos-2 human osteosarcoma cells. BacMam viruses containing full-length cDNAs were constructed for both human ER subtypes, ERalpha and ERbeta, and a third BacMam virus containing an ER-responsive reporter gene cassette. Using these viruses, we found that BacMam-ER expression/reporter constructs could be used to profile the effects of the agonist 17beta-estradiol and the partial agonist raloxifene in human Saos-2 cells. A comparison of assay data obtained with the BacMam-based system with that using standard DNA transfections demonstrates that the two systems are functionally equivalent, giving comparable EC50 and IC50 values for estrogen and estrogen plus raloxifene treatments, respectively. Our results indicate that BacMam-mediated gene transfer offers a novel and efficient method for delivery of nuclear receptors and associated genes for mammalian cell-based assay development.