Stephanou A, Handwerger S
Division of Endocrinology, Children's Hospital Medical Center, University of Cincinnati College of Medicine, OH.
Biochem Biophys Res Commun. 1995 Jan 5;206(1):215-22. doi: 10.1006/bbrc.1995.1030.
Transient transfection studies using deletion mutants of the hPL promoter indicate that the DNA elements for NF-IL6 responsiveness are located between -2.3 to -1.1 kb. Subsequent transfection studies using a hPL promoter fragment containing the region between -1376 to -1088 bp ligated to a heterologous SV40 CAT vector (NF-IL6/hPL-CAT) demonstrated that the NF-IL6/hPL-CAT construct is responsive to NF-IL6. Mobility shift assays using nuclear extracts from BeWo choriocarcinoma cells overexpressing NF-IL6 demonstrated specific binding of the extracts to a labeled oligonucleotide probe to this region of the hPL promoter. These studies therefore strongly suggest that the effect of IL-6 on hPL gene expression is mediated, at least in part, by the binding of NF-IL6 to a region of the hPL promoter that contains three NF-IL6 responsive elements.
使用人胎盘催乳素（hPL）启动子缺失突变体进行的瞬时转染研究表明，NF-IL6反应性的DNA元件位于-2.3至-1.1 kb之间。随后使用连接到异源SV40 CAT载体（NF-IL6/hPL-CAT）的包含-1376至-1088 bp区域的hPL启动子片段进行的转染研究表明，NF-IL6/hPL-CAT构建体对NF-IL6有反应。使用过表达NF-IL6的BeWo绒毛膜癌细胞的核提取物进行的凝胶迁移试验表明，提取物与hPL启动子该区域的标记寡核苷酸探针有特异性结合。因此，这些研究强烈表明，IL-6对hPL基因表达的影响至少部分是由NF-IL6与hPL启动子中包含三个NF-IL6反应元件的区域结合介导的。
