PLACENTAL/UTERINE EFFECTS OF CHLORINATED HYDROCARBONS

这项拟议的研究将调查 氯代烃2，3，7，8-四氯二苯并对二恶英和 多氯联苯对胎盘有破坏作用- 子宫功能。假设TCDD和多氯联苯改变胎盘- 子宫旁分泌和自分泌网络对 滋养层细胞的侵袭性、增殖和激素分泌 随着子宫细胞的生长。主要的网络包括表皮生长。 转化生长因子(EGF)及其受体 (TGFs)-α和-β、细胞因子IL-1B和 蛋白水解酶抑制物、纤溶酶原激活物抑制物(PAI)。印刷电路板 同系物3，3‘，4，4’-四氯和2，2‘，4，4’，5，5‘-六氯，以及 将对阿罗氯1254混合物进行研究，以代表不同水平的 啊，诱导活性。 特定目的#1：培养的人胎盘滋养层细胞的暴露 细胞系对TCDD和多氯联苯改变重要基因的表达 胎盘生长和内分泌功能？实验将具体地 评估：A)绒毛膜癌中EGF受体结合和激酶活性 确定JEG-3和BeWo细胞是否存在下调 细胞增殖或激素分泌改变的受体；b) 在其他人类细胞系中被认为受TCDD调控的基因， 包括转化生长因子-α、转化生长因子-β、白介素1β和纤溶酶原激活物；c)诱导 细胞色素P-450 1A1和多氯联苯代谢，以确定是否有剂量- 与EGF受体和/或生长变化平行的相关效应 通过ah受体表达的因子。 特定目的#2：培养的人子宫内膜暴露 腺癌细胞对多氯联苯和TCDD表达的影响 调控基因？实验将评估：a)EGF-R结合和激酶 活性与EGF刺激的细胞增殖；b)雌激素受体 以确定细胞对雌激素的反应是否 改变；c)转化生长因子-α、转化生长因子-β、IL-1B和纤溶酶原激活物的表达；d) 诱导细胞色素P450 1A1和多氯联苯代谢的剂量效应 与上述a)、b)和c)的关系。 特定目的#3：在怀孕的大鼠中，TCDD和 不同的多氯联苯改变胎盘P450 1A1、TGFs基因的表达- 胎儿胎盘生长与α-β、IL-1B、PAI的关系 智障？实验将评估：a)胎儿-胎盘生长晚期 B)转化生长因子-α、转化生长因子-β、白介素1β和纤溶酶原激活物的表达 胎盘和蜕膜；c)细胞色素P450 1A1诱导和多氯联苯代谢 与上述a)和b)的剂量反应关系。 特异性目的4：胎盘氨基酸转运系统的表达 妊娠胎盘发育迟缓与胎盘发育异常的相关性研究 用多氯联苯和TCDD治疗的大鼠？A)通过系统运输氨基酸 B、+、y+、XAG-和A将在以下分离的囊泡制剂中进行检测 将检测微绒毛和基底膜；b)信使核糖核酸水平 利用cDNA探针构建Y+和XAG-系统；c)细胞色素P450 1A1 对上述a)和b)的DoS-响应关系进行归纳。

The proposed research will investigate mechanisms by which the chlorinated hydrocarbons 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and polychlorinated biphenyls (PCBs) have disruptive effects on placental- uterine function. The hypothesis is that TCDD and PCBs alter placental- uterine paracrine and autocrine networks which are important for trophoblast invasiveness, proliferation and hormone secretion, as well as uterine cell growth. The major networks involve epidermal growth factor (EGF) and its receptor (EGF-R), transforming growth factors (TGFs)-alpha and -betas, the cytokine interleukin (IL) -1B and the protease inhibitor, plasminogen activator inhibitor (PAI). The PCB congeners 3,3',4,4'-tetrachloro and 2,2',4,4',5,5'-hexachloro, and the mixture Arochlor 1254 will be studied to represent different levels of AHH induction activity. SPECIFIC AIM #1: Does exposure of cultured human placental trophoblstic cell lines to TCDD and PCBs alter expression of genes important for placental growth and endocrine function? Experiments will specifically evaluate: a) EGF receptor binding and kinase activity in choriocarcinoma JEG-3 and BeWo cell lines to determine whether there is down-regulation of receptors with altered cell proliferation or hormone secretion; b) The genes recognized to be regulated by TCDD in other human cell lines, including TGF-alpha, TGF-betas, IL-1B, and PAI; c) Induction of cytochrome P-450 1A1 and PCB metabolism to determine if there are dose- related effects in parallel with changes in EGF receptors and/or growth factor expression through the Ah receptor. SPECIFIC AIM #2: Does exposure of cultured human endometrial adenocarcinoma cells to PCBs and TCDD alter expression of growth regulatory genes? Experiments will evaluate: a) EGF-R binding and kinase activity and EGF-stimulated cell proliferation; b) Estrogen receptor levels to determine whether the cellular response to estrogens is altered; c) Expression of TGF-alpha, TGF-betas, IL-1B, and PAI; d) Induction of cytochrome P450 1A1 and PCB metabolism for dose-response relationships with a), b) and c) above. SPECIFIC AIM #3: In pregnant rats, does administration of TCDD and the respective PCBs alter expression of placental genes for P450 1A1,TGFs- alpha and -beta, IL-1B and PAI in correlation with feto-placental growth retardation? Experiments will evaluate: a) Feto-placental growth in late gestation; b) Expression of TGF-alpah, TGF-betas, IL-1B, and PAI in placenta and decidua; c) Cytochrome P450 1A1 induction and PCB metabolism for dose response relationships with a) and b) above. SPECIFIC AIM #4: Is expression of placental amino acid transport systems altered in correlation with feto-placental growth retardation in pregnant rats treated with PCBs and TCDD? a) Amino acid transport via Systems B,+,y+,XAG-, and A will be examined in isolated vesicle preparations from the microvillous and basal membranes; b) mRNA levels will be examined utilizing cDNA probes to Systems y+ and XAG-; c) Cytochrome P450 1A1 induction for dos-response relationships with a) and b) above.