The mechanism of exocrine gland disorder through estrogen and/or endocrine disruptors

为了研究雌激素和内分泌干扰物对EBY再激活的可能性，我们分析了BZLF 1启动子测定和ZEBRA蛋白的免疫印迹。在B 95 -8和Akata细胞中，Escherichia coli抑制TPA诱导的ZEBRA表达。建立了稳定转染pZp 221-Luc和pZp 552-Luc的B 95 -8细胞系。在这些细胞中，雌二醇也抑制TPA激活的荧光素酶活性。在Hela、HSY和HSG细胞中，雌二醇抑制ZEBRA转染或TPA刺激诱导的Zp活性。雌激素的作用在30分钟内就显示出来，因此我们推测这些作用的一部分可能是通过膜ER或GPR 30的非基因组事件。雌激素对Zp活性的作用机制正在详细研究中。接下来，我们分析了内分泌干扰物的影响与相同的程序使用BPA，3-MC，B[a]P和TCDD。这些化学物质都没有改变B 94 -8和Akata细胞中的ZEBRA表达或Zp活性。与对B细胞的这些作用相反，3-MC和TCDD在Hela、HSG和HSY细胞中增强Zp活性。与单独转染AhR/Arnt相比，转染AhR/Arnt和ER后，Zp在Hela细胞中的活性降低，而在HSG和HSY细胞中的活性增强。这些结果表明，雌激素受体可以调节Zp活性的二恶英诱导依赖于细胞类型。在HSY细胞中，虽然ZEBRA反式激活Zp没有改变二恶英无AhR/Arnt，Zp的高活性时，观察到ZEBRA和AhR/Arnt共转染。我们推测，二恶英可能是一种增强剂的EBV再激活上皮细胞。为了确定AhR/Arnt与ZEBRA的相互作用，我们使用免疫沉淀、GST pull-down分析、凝胶迁移分析和足迹法进行了研究。

To study the possibility of EBY reactivation by estrogen and endocrine disruptors, we analyzed BZLF1 promoter assay and immunoblotting for ZEBRA protein. Estradiol inhibited ZEBRA expression induced by TPA in B95-8 and Akata cells. We established stable B95-8 cell lines which transfected pZp221-Luc and pZp552-Luc. In these cells estradiol also inhibited luciferase activity activated by TPA. In Hela, HSY and HSG cells, estradiol inhibited Zp activity induced by both ZEBRA transfection or TPA stimulation. The effect of estrogen was revealed in 30 minutes, thus we speculated a part of these effects might be a non-genomic event through membrane ER or GPR30. The mechanisms of estrogen on Zp activity are being searched in detail. Next, we analyzed the effects of endocrine disruptors with the same procedure using BPA,3-MC,B[a]P and TCDD. None of these chemicals changed the ZEBRA expression or Zp activity in B94-8 and Akata cells. In contrast to these effects on B cells, Zp activity was enhanced by 3-MC and TCDD in Hela, HSG and HSY cells. When we transfected AhR/Arnt and ER, the activity of Zp was decreased in Hela cells whereas enhanced in HSG and HSY cells compared with AhR/Arnt alone. These results indicated that ER could regulate the Zp activity induced by dioxins depending on the cell types. In HSY cells, although ZEBRA transactivation of Zp was not changed by dioxins without AhR/Arnt, high activity of Zp was observed when ZEBRA and AhR/Arnt were cotransfected. We speculated that dioxins might be an enhancer of EBV reactivation in epithelial cells. To determine the interaction between AhR/Arnt and ZEBRA, we are investigating using immunoprecipitation, GST pull-down assay, Gel shift assay and foot printing.