Li Z L, Abe H, Ueki K, Kumagai K, Araki R, Otsuki Y
Department of Anatomy, Osaka Medical College, Osaka, Japan.
J Histochem Cytochem. 2003 Dec;51(12):1601-9. doi: 10.1177/002215540305101204.
We describe the application of the biomolecular interaction (BIA) technique to detection of the interaction between protein (e.g., c-Jun) and DNA (e.g., two AP-1 motifs from bcl-2 promoter), compared with immunohistochemistry (IHC) of c-Jun. The specific binding assay for the interaction of c-Jun and activating protein-1 (AP-1) motifs was performed using a Biacore 2000 system. Intense immunoreactivity of c-Jun in glandular cells of the human uterine endometrium was observed in the proliferative phase, while c-Jun in stromal cells was expressed throughout the menstrual cycle. In contrast to the IHC of c-Jun, the specific binding of c-Jun to two separate AP-1 motifs in the bcl-2 promoter region was detected only in nuclear extracts of glandular cells, but not in stromal cells, during the proliferative phase. These results indicate that, while transmitting various signals, c-Jun enhances the transcription level of bcl-2, which in turn keeps glandular cells alive and proliferating in normal human endometrium during the proliferative phase. Moreover, the method involving real-time biomolecular interactions such as DNA-protein binding is novel for the study of transcription factors when combined with IHC.
我们描述了生物分子相互作用（BIA）技术在检测蛋白质（如c-Jun）与DNA（如来自bcl-2启动子的两个AP-1基序）之间相互作用中的应用，并与c-Jun的免疫组织化学（IHC）进行了比较。使用Biacore 2000系统对c-Jun与活化蛋白-1（AP-1）基序的相互作用进行了特异性结合测定。在增殖期观察到人类子宫内膜腺细胞中c-Jun有强烈的免疫反应性，而基质细胞中的c-Jun在整个月经周期均有表达。与c-Jun的免疫组织化学不同，仅在增殖期的腺细胞核提取物中检测到c-Jun与bcl-2启动子区域中两个独立的AP-1基序的特异性结合，而在基质细胞中未检测到。这些结果表明，c-Jun在传递各种信号的同时，增强了bcl-2的转录水平，进而在增殖期使正常人类子宫内膜中的腺细胞存活并增殖。此外，当与免疫组织化学结合时，涉及DNA-蛋白质结合等实时生物分子相互作用的方法对于转录因子的研究来说是新颖的。
