Druege P M, Klein-Hitpass L, Green S, Stack G, Chambon P, Ryffel G U
Nucleic Acids Res. 1986 Dec 9;14(23):9329-37. doi: 10.1093/nar/14.23.9329.
We introduced estrogen responsiveness as a new characteristic into rat hepatoma, mouse Ltk- and human HeLatk-cells by transfecting the human estrogen receptor (ER) cDNA. To measure the estrogen response we used Xenopus vitellogenin gene A2 constructs linked to the bacterial CAT gene. Transient cotransfections of the ER cDNA and the vitellogenin gene-CAT constructs containing the estrogen responsive element (ERE) lead to a hormone dependent induction of CAT activity whereas cotransfected vitellogenin gene constructs lacking the ERE are not inducible. Stable transfections of ER cDNA into Ltk- cells give rise to cell clones that are estrogen responsive as shown by transfection of various vitellogenin gene-CAT constructs. These results prove that the transfected ER is biologically active and is sufficient to make a cell estrogen responsive.
我们通过转染人雌激素受体（ER）cDNA，将雌激素反应性作为一种新特性引入大鼠肝癌细胞、小鼠Ltk - 细胞和人HeLa tk - 细胞。为了测量雌激素反应，我们使用了与细菌氯霉素乙酰转移酶（CAT）基因相连的非洲爪蟾卵黄蛋白原基因A2构建体。ER cDNA与含有雌激素反应元件（ERE）的卵黄蛋白原基因 - CAT构建体的瞬时共转染导致CAT活性的激素依赖性诱导，而缺乏ERE的共转染卵黄蛋白原基因构建体则不可诱导。将ER cDNA稳定转染到Ltk - 细胞中产生了细胞克隆，如通过转染各种卵黄蛋白原基因 - CAT构建体所示，这些克隆对雌激素有反应。这些结果证明，转染的ER具有生物学活性，并且足以使细胞对雌激素产生反应。
