人类绒毛膜癌细胞JEG-3论文：四种手性农药对甾体类激素代谢影响的对映体选择性

1、人类绒毛膜癌细胞 JEG-3 论文： 四种手性农药对甾体类激素 代谢影响的对映体选择性【中文摘要】现代社会中手性农药的使用越来越普遍。 本文考察了 4 种典型手性农药(o,p -DDT 三氯杀虫酯、氯菊酯和对溴磷)外 消旋及其对映体对内分泌相关基因的干扰情况。探讨了以人类绒毛膜癌 JEG-3 细胞为模型，研究了这些手性农药外消旋及其对映体暴露对 产妇胎儿健康的潜在风险。研究通过采用实时定量PCR 勺方法,发现这些典型手性农药的外消旋及其对映体对促性腺释放激素(GnRHI,GnRHE )及其受体(GnRHR)胆固醇来源的雌激素合成以及胎儿在 怀孕期间母体免疫的相关基因(HLA-G)表达都有不同程

2、度的影响。实 验结果显示4种手性农药的对映体对这些内分泌干扰相关基因的干 扰影响的大小分别为:R-( - )-o,p -DDTS-(+)-o,p -DDT;S-(+)-AFR-( - )-AF;1R-cis-PM 和 1S-trans-PM 处理组大于1S-cis-PM 和 1R-trans-PM;(-)-Leptophos(+)-Leptophos(存在统计学意义上的显著性差异，pv0.05)。通过 o,p -DDT 的实验结果表 明,手性污染物对产生类固醇甾类基因的影响不仅仅是对雄性甾类激 素或者雌性甾类激素影响，同时也干扰其他体内的激素分泌。值得注 意的是,AF外消旋及其对映体的结果说明

3、了外消旋化合物与其对映体 对各个产生甾类激素的相关基因的影响不是简单的相加效应；(-)-Leptophos 相对于什)-Leptophos 具有更强的内分泌干扰效应。 首次有数据说明了哺乳动物和水生动物对手性农药对溴磷的对应体选择性是有差异的。这为今后纯手性新型的农药的开发和研究提供了 一定的参考价值。 o,p -DDT 及其对映体对 MCF-7 乳腺癌细胞侵润相 关基因，基质金属蛋白酶基因(MMP-2 和 MMP-9 和组织基质金属蛋白 酶抑制因子基因(TIMP1、3、4)的 mRNA 勺表达具有显著性的上调作用，同时也能够促进该癌细胞的侵润转移能力的增加。并且数据表明 R-( - )-o,

4、p -DDT与 S-(+)-o,p -DDT 处理组对侵润相关基因和侵 润转移能力的选择性均有统计学意义;o,p -DDT 及其对映体很有可 能同时通过 ER 介导和非经典的途径来作用于细胞侵润相关酶类，从 而影响 MCF-7 细胞侵润能力。这为今后纯手性新型的农药的开发和研 究提供了一定的参考价值。【英文摘要】 Chiral pesticides are used more and more popular. Inthis study, we employed typical chiral pesticides as model compo un ds,JEG-3 cell li nes a

5、s in vitro model and assessed enan tioselective disruption of these chiral pestisides towards the metabolic pathway ofGnRH-mediated and estroge n derived from cholesterol syn thesis.Besides, we desired to evaluate the pote ntial risks of these compo undsto maternal-fetal health. The results of real-

6、time PCR revealed that these4 typical chiral pesticides exhibited strong in flue nce on these gen es, includi ng GnRH and its receptor, genes corresponding to enzymes of thesteroidogenesispathwayand HLA-G. The enan tioselective pote ntial of en docri nedisruption effect caused by the four chiral pes

7、tisides to these genes wereas follows: R-(- )-o,p -DDT S-(+)-o,p -DDT,S-(+)-AF R-( - )-AF. The groups of 1R-cis-PM and 1S-trans-PM 1S-cis-PM and 1R-tra ns-PM, (-)-Leptophos (+)-Leptophos(p0.05).The results of o,p -DDTindicated that the effection of chiral contam inants were n ot merely on an droge n

8、 steroid hormones or estrogensteroid hormones, but also showeddisorder of endocrine regulation invivo. Data of AF concerning on the steroidogenesis genesindicated thattwo enantiomers were notacted as a simple additi on whe n mixed as equalmole. The en docri nedisrupti ng effects of (-)-Leptophos was

9、 much stron ger tha n(+)-Leptophos. To my best of my kno wledge, our data firstly in dicatedthat the enan tioseletive effect ofmammaand aquatic animal are different. o,p -DDT and its twoenantiomers exhibited significant up regulation on matrix metalloproteinases genes (MMP-2 and MMP-9) and tissue in hibitorofmetalloprote in asesgenes (TIMP 1, 3, 4). Besides,they also had the pote ntial to promote in vasive ness of breast cancer cell.Whaf s more, our data showedthat the disturbanee of o,p -DDT on