Observation of mechanisms for transcriptional regulation

1.通过化学发光或荧光免疫组织化学方法，在3 -甲基胆蒽刺激下，H4IIE细胞系和肝细胞切片的HSP90在细胞质中与芳烃受体（AhR）分离，但在细胞核中未分离。然而，FLET分析显示，这两种蛋白都与松散连接或互连因子相关。我们应用这项技术来检测一种新的蛋白的细胞内定位，这种蛋白只定位于内分泌细胞。通过高灵敏度、定量的CYP2B2 mRNA原位杂交和er3体视学分析，发现正常和苯巴比妥治疗大鼠肝细胞中CYP2B2的表达与内质网（ER）面积密切相关。当一个报告基因连接到CYP1A1基因的5'-区域时，我们可以通过光子成像检测该基因的定位。然而，在转录因子Sp1或CBP/p-300存在的情况下，通过光子检测和/或原子力显微镜观察，CYP1A1基因5 '区和AhR-Arnt的DNA结构没有明显差异。

1.By the chemiluminescent or fluorescent immunohistochemistry, HSP90 localized with arylhydrocarbon receptor(AhR) was dissociated from AhR in the cytoplasm but not in the nuclei after the stimulation of 3 -methylcholanthrene in the H4IIE cell lines and hepatocytes in sections from rat livers. However, FLET analysis revealed that the both proteins associated with a loose connection or with interconnecting factor(s). We applied this technique to examine the intracellular localization of a novel protein that localized exclusively in the endocrine cells.2.Aclose relationship was found between the expression of CYP2B2 and area of endoplasmic reticulum(ER) in hepatocytes from normal and phenobarbital-treated rats as revealed by a combination of highly-sensitive and quantitative in situ hybridization of CYP2B2 mRNA with stereological analysis of ER.3.When a reporter gene was connected to 5'-region of CYP1A1 gene, we could detect the localization of the gene by the photon imaging. However, no significant difference was found in the DNA structure if 5 '-region of CYP1A1 gene and AhR-Arnt were reacted under the presence of transcriptional factors, Sp1 or CBP/p-300 by the photon detection and/or atomic-force microscopy.