用MCF-7细胞检测有机磷农药拟雌激素样活性

目的 观察常用有机磷农药对MCF-7细胞增殖以及对雌激素受体基因和pS2基因表达的影响。方法 选用乐果、乙酰甲胺磷、久效磷、马拉硫磷、对硫磷、对氧磷6种有机磷农药进行MCF-7人乳腺癌细胞增殖实验和转录活化实验。结果 6种有机磷农药不能诱导MCF-7人乳腺癌细胞增殖,但乐果却能使pS2基因表达上调。结论 乐果可能具有拟雌激素样活性,且引起pS2基因表达上调不通过雌激素受体介导。     

有机磷农药环境雌激素样的作用

目的 初步探索几种常用有机磷农药的环境雌激素样作用,为有机磷农药对食品污染的综合防治提供依据。方法通过MCF-7人乳腺癌细胞增殖试验,对乐果(Dimethoate)、对硫磷(Parathion)、对氧磷(Paraoxon)3种有机磷农药进行雌激素样活性研究。结果 乐果10-5 mol·L-1、10-6mol·L-1剂量组对MCF-7乳腺癌细胞的增殖指数分别为1.31±0.07、1.56±0.08,较对照组的1.00±0.14有显著差异(P<0.05);对硫磷在10-6mol·L-1、10-7mol·L-1剂量组对MCF-7细胞的增殖指数分别为1.35±0.02、1.39±0.09,与对照组相比也有显著差异(P<0.05)。结论 乐果、对硫磷在适宜剂量范围内有拟雌激素样作用。     

竞争性受体结合试验检测农药拟雌激素样作用

【目的】应用竞争性受体结合试验检测常用六种农药的雌激素样活性。[方法]选用乐果、乙酰甲胺磷、久效磷、马拉硫磷、对硫磷和对氧磷6种有机磷农药进行大鼠子宫雌激素受体竞争性结合试验。【结果】应用多点饱和分析建立Scatchard图，采用10nmol／L^3 H-E。作单点饱和分析，结果显示这6种有机磷农药的结合率均未下降到50％。【结论】这6种有机磷农药不能与大鼠子宫雌激素受体的位点结合。     

邻苯二甲酸二丁酯的拟雌激素活性研究

目的探讨邻苯二甲酸二丁酯(DBP)的拟雌激素活性。方法对雌激素敏感的人乳腺癌MCF-7细胞在RPMI 1640培养基中采用开放式单层贴壁培养,开始实验前将细胞用PBS洗涤后改为在无酚红RPMI 1640培养基中培养5d,实验设溶剂对照、雌激素阳性对照和邻苯二甲酸二丁酯各剂量组(10^-7-10^-3mol／L),采用四唑盐(MTT)法、生长曲线、有丝分裂指数和细胞克隆形成试验对MCF-7细胞增殖情况进行分析。结果与溶剂对照组相比,10^-5mol／LDBP处理24h就可促进MCF-7细胞增殖,提高增殖指数;随着培养时间延长至96h,其他浓度DBP也表现出促进细胞增殖的效果,且浓度在10^-5mol／L时,细胞增殖活性达到最大;在对数生长期,DBP可提高MCF-7细胞的有丝分裂指数;10^-5mol／LDBP处理48h就可增强MCF-7细胞形成细胞克隆的能力。结论邻苯二甲酸二丁酯可促进雌激素依赖性乳腺癌MCF-7细胞的增殖,可能具有拟雌激素作用。     

48例拟除虫菊酯类农药中毒临床分析

拟除虫菊酯是目前在农业生产中应用最多、最广的一种新型农药,是在天然除虫菊酯化学结构的基础上开发的新型农药。农业上常用的有溴氰菊酯(敌杀死)氯氰菊酯(功天)氰戊菊酯(速灭杀丁)等等。由于农业上的广泛应用,中毒者常见。现总结1996～1998年我院收治的拟除虫菊酯类农药48例报告如下:1 一般资料 本组溴氰菊酯27例,氯氰菊酯13例,氰戊菊酯8例;中毒途径口服43例,皮肤接触5例;男19例,女29例,年龄14～59岁,服毒量1～5ml 7例,6～10ml 21例10～40ml 3例,40～100ml 5例,100ml以上4例,不详8例;口服至就诊时间1小时以内     

用MCF-7细胞检测多氯联苯拟雌激素样活性

目的观察多氯联苯（PCB）对MCF-7细胞增殖及生长曲线的影响。方法选用多氯联苯（1254）进行MCF-7人乳腺癌细胞增殖实验。结果PCB组细胞增殖指数均明显高于溶剂对照组，10ng／LPCB可诱导MCF-7细胞最大增殖（P〈Q01）。生长曲线分析也显示，PCB、E2生长曲线明显上移，有明显刺激MCF-7细胞增殖的类雌激素活性。结论提示PCB具有拟雌激素样活性。     

顺式和反式氯氰菊酯拟雌激素样作用探讨

目的通过体内和体外实验相结合的方法,探讨顺式氯氰菊酯和反式氯氰菊酯是否具有拟雌激素样作用。方法进行去势大鼠子宫增重实验、小鼠子宫增重实验和乳腺癌细胞株(MCF-7)细胞增殖实验。结果两种子宫增重实验中两种受试物均未观察到子宫质量增加,而阳性对照组可见到明显子宫增重作用。两种受试物在10-8~10-10mol/L均导致MCF-7细胞明显增殖,与溶剂对照组比较均有统计学意义(P<0.05)。结论两种受试物在体内实验中未见明显雌激素样作用;在体外细胞实验中似乎具有拟雌激素样作用。     

染料木黄酮对人乳腺癌细胞雌激素样作用机制研究

目的 探讨染料木黄酮(genistein,GEN)对人乳腺癌细胞增殖影响及作用机制.方法 MTT法测定GEN对雌激素依赖性乳腺癌细胞MCF-7和非雌激素依赖性乳腺癌细胞MDA-MB231增殖作用,应用雌激素受体拮抗剂IC1182780观察GEN的雌激素样作用途径,采用流式细胞术分析乳腺癌细胞周期情况.结果 GEN可促进MCF-7细胞的增殖,使G1期向S期转变,增殖作用可被雌激素受体拮抗剂所拮抗;对非雌激素依赖性MDA-MB231细胞增殖无影响.结论 GEN通过雌激素受体(ER)介导而发挥雌激素样活性作用.     

甘草次酸和甘草甜素雌激素样作用研究

目的探讨甘草次酸和甘草甜素两种三萜类成分的雌激素样作用,为开发新的活性植物雌激素成分奠定基础。方法采用溴化噻唑蓝四氮唑(MTT)法观察两种三萜成分对雌激素受体(ER)阳性人乳腺癌细胞系MCF-7细胞增殖的影响,Western Blot法和半定量RT-PCR法测定两种三萜成分对MCF-7细胞ER亚型蛋白及mRNA表达的影响。实验数据(±s)采用SPSS 23.0进行统计分析,基于单因素方差分析的方法,组间比较用SNK法和LSD法。结果 100μmol/L和10μmol/L甘草甜素能够抑制MCF-7细胞的增殖;100μmol/L、1μmol/L和10-2μmol/L甘草次酸能够促进MCF-7细胞的增殖;10μmol/L的甘草次酸可诱导MCF-7细胞ERα、ERβ蛋白表达,mRNA表达均显著升高(P0.05);加入1×10~(-7)μmol/L的雌激素受体拮抗剂ICI182780均能抑制甘草次酸的以上效应。结论甘草次酸具有促进MCF-7细胞增殖作用,其作用机制可能是上调MCF-7细胞ERα、ERβ基因和蛋白的表达,显示有雌激素样作用,且其雌激素样作用可能是ER介导产生的。抗雌激素样作用的甘草甜素的作用机制还有待于深入研究。     

杀螟硫磷体外环境雌激素活性及机制研究

目的 检测杀螟硫磷的环境雌激素活性，并探讨其作用机制。方法 人乳腺癌细胞株（MCF-7）细胞在含10％小牛血清的RPMI1640培养基中进行半开放式贴壁培养，加受试物前2周，改在含10％去激素小牛血清的无酚红RPMI1640培养基中培养。试验设溶剂对照组、阳性对照组及杀螟硫磷5个不同浓度实验组，采用MCF-7细胞增殖试验、四甲基偶氮噻唑蓝（MTT）试验和生长曲线观察杀螟硫磷对MCF-7细胞增殖的影响，并通过雌激素受体（ER）结合试验分析杀螟硫磷与ER的结合情况。结果 杀螟硫磷在10^-9～10^-6mol／L剂量范围内可诱导MCF-7细胞增殖，差异有统计学意义（P〈0．05），且存在剂量-效应关系。杀螟硫磷能与ER结合，抑制^125I-E2与ER结合。结论 杀螟硫磷具有环境雌激素活性，其拟雌激素作用由ER介导而产生。     

汞、铬和锰化合物雌激素样作用的实验研究

目的为评价汞、铬和锰化合物雌激素样作用。方法选用氯化汞、硫酸锰、氯化铬、三氧化铬4种化合物进行MCF7人乳腺癌细胞增殖试验和雌激素受体竞争结合试验。结果10-9×10-5molL氯化汞能诱导MCF7人乳腺癌细胞增殖,10-7molL氯化汞使MCF7人乳腺癌细胞增殖达最大,其增殖作用可被雌激素受体拮抗剂ICI182780完全阻断;但不影响雌二醇与雌激素受体结合。硫酸锰、氯化铬、三氧化铬均不能诱导MCF7人乳腺癌细胞增殖,且不影响雌二醇与雌激素受体结合。结论氯化汞可能通过雌激素受体介导而表现出雌激素样作用;硫酸锰、氯化铬、三氧化铬不具有雌激素样作用。     

The implementation of a battery of in vivo and in vitro bioassays to assess river water for estrogenic endocrine disrupting chemicals

Previous research has shown that accurate evaluation of environmental water samples for estrogenic activity requires a panel of in vitro and in vivo bioassays, which are based on different molecular and cellular action mechanisms. In the current study, a test battery containing four assays was used to analyze water from the Eerste River, South Africa for estrogenicity. Three sites were used for analysis, namely Jonkershoek (control site situated in the mountains at the origin of the Eerste River), sewage effluent from Stellenbosch sewage treatment works and Spier site (sampling site on the Eerste River downstream from Stellenbosch). Estrogenicity was determined using an estrone enzyme linked immuno sorbent assay (ELISA), estrogen induced proliferation of human breast cancer adenocarcinoma cells (MCF-7) also known as the E-SCREEN, estrogen induced suppression of estrogen receptor alpha protein expression (ER-α) in MCF-7 cells (ERα assay) and by monitoring estrogen induced vitellogenin (VTG) synthesis in juvenile Oreochromis mossambicus (VTG assay). Low concentrations of estrone (ranging between 1.4 and 2.2 ng/l) near the detection limit of the assay were detected in samples collected from Jonkershoek. Water from this site shows no estrogenicity in the E-SCREEN, ERα assay or VTG synthesis bioassay. The estrone concentrations in the sewage effluent extracts, as well as Spier site extracts, ranged between 14.7 and 19.4 ng/l. The assays using ERα induction by the MCF-7 cell line, MCF-7 proliferation and in vivo VTG synthesis by juvenile tilapia showed that these samples are estrogenic. The results obtained for the assays in the battery are comparable.     

A novel endocrine-disrupting agent in corn with mitogenic activity in human breast and prostatic cancer cells

Housing adult rats on ground corncob bedding impedes male and female mating behavior and causes acyclicity in females. The suppressive effects on ovarian cyclicity are mimicked by a mitogenic agent purified from the ground corncob bedding material (corn mitogen; CM), which stimulates the proliferation of estrogen receptor (ER)-positive (MCF-7 cells) and ER-negative (MDA-MD-231 cells) breast cancer cells. Purified CM does not compete for [(3)H]estradiol binding to ER or nuclear type II sites, and its effects on MCF-7 breast cancer cell proliferation are not blocked by the antiestrogen ICI-182,780. These results suggest that the active component is unlikely to be a phytoestrogen, bioflavonoid, mycotoxin, or other known endocrine-disrupting agent that modifies cell growth via ER or type II [(3)H]estradiol binding sites. CM also stimulates the proliferation of PC-3 human prostatic cancer cells in vitro, and the growth rate of PC-3 cell xenografts is accelerated in nude male mice housed on ground corncob as opposed to pure cellulose bedding. Consequently, this endocrine-disrupting agent in ground corncob bedding may influence behavioral and physiologic reproductive response profiles and malignant cell proliferation in experimental animals. Fresh corn (kernels and cob) or corn tortillas also contain CM, indicating that human exposure is likely; consequently, CM and/or related mitogens in corn products may influence human health and development.     

二氯二苯二氯乙烯的类雌激素活性的实验研究

目的研究二氯二苯二氯乙烯(DDE)的类雌激素活性对人乳腺癌细胞株MCF-7增殖的影响。方法采用了体外MCF-7细胞增殖试验检测了DDE(3×10-5、3×10-6、3×10-7、3×10-8、3×10-9 mol/L)的类雌激素活性,并用生长曲线分析对其作用机制进行了初步探讨。结果3×10-6 mol/LDDE染毒组吸光度值高于溶剂对照组且MST-7细胞增殖的生长曲线与10-9 mol/L17β-雌二醇染毒组相似。结论DDE具有刺激MCF-7细胞增殖的类雌激素活性,其机制可能与17β-雌二醇相同。     

Specific in vitro toxicity of crude and refined petroleum products: 3. Estrogenic responses in mammalian assays

Current petroleum risk assessment considers only narcosis as the mode of action, but several studies have demonstrated that oils contain compounds with dioxin-like, estrogenic or antiestrogenic, and androgenic or antiandrogenic activities. The present study is the third in a series investigating the specific toxic effects of 11 crude oils and refined products. By employing recombinant mammalian cells stably transfected with the human estrogen receptor alpha (ERα) or beta (ERβ), and expressing the luciferase protein (ERα-U2OS-Luc and ERβ-U2OS-Luc assay), the estrogenicity or antiestrogenicity of oils was studied. All oils, except for two refined oils and one crude oil, induced estrogenic responses. The calculated estrogenic potencies of the oils were six to nine orders of magnitude lower than the potency of 17β-estradiol (E2). Upon coexposure to a fixed concentration of E2 and increasing concentrations of oils, additive, antagonistic, and synergistic effects were revealed. One nautical fuel oil was tested in the human breast carcinoma cell line MCF-7, in which it induced cell proliferation up to 70% relative to the maximal induction by E2. At its minimum effect concentration of 25?mg/L, the oil was also capable of inducing mRNA expression of the estrogen-dependent protein pS2 by a factor of two. The present results indicate that oils naturally contain potentially endocrine-disrupting compounds that are able to influence the estrogenicity of other compounds and may cause biological responses beyond receptor binding.     

氯化汞雌激素样作用及其机制的实验研究

目的评价氯化汞(HgCl2)的雌激素样作用及其作用机制.方法从体内和体外两个水平,观察不同浓度的HgCl2对MCF-7人乳腺癌细胞增殖作用、去卵巢SD大鼠子宫的诱导增生作用、过氧化物活力的变化及对雌激素结合雌激素受体的影响.结果 10-6～10-9 mol/L的HgCl2均可引起MCF-7人乳腺癌细胞增殖,10-7 mol/L的HgCl2使MCF-7人乳腺癌细胞增殖达最大.每天0.04、0.20及1.00 mg/kg连续3 d腹腔注射HgCl2能刺激大鼠子宫增生和过氧化物酶活力增加;但HgCl2不影响雌二醇(E2)与雌激素受体结合.结论 HgCl2可能通过雌激素受体介导而表现出雌激素样作用,但不与E2竞争结合雌激素受体.     

有机磷农药与再生障碍性贫血发病关系的Meta分析

目的 评价有机磷农药与再生障碍性贫血发病的关系,为一级预防提供科学依据。方法 检索1990年1月至2014年8月公开发表在中国生物医学文献数据库(CBM)、中国知网(CNKI)、PubMed和EMBASE关于有机磷农药与再生障碍性贫血发病关系的病例对照研究。制定纳入和排除标准,对符合纳入标准的文献进行质量评价,利用Review Manager 5.0软件计算有机磷农药的合并OR值及其95%CI,并对文献的研究人群和有机磷农药使用方式进行亚组分析。结果 最终共纳入9篇文献,5 833例研究对象,其中病例组1 404人,对照组4 429人。结果显示有机磷农药可增加再生障碍性贫血的发病风险(OR=1.97,95%CI:1.60～2.44)。亚组分析显示亚洲人群再生障碍性贫血的发病风险(OR=2.01,95%CI:1.52～2.66)高于欧美人群(OR=1.93,95%CI:1.39～2.67);使用单一有机磷类农药(OR=2.15,95%CI:1.60～2.88)比使用有机磷类农药的混配制剂(OR=1.82,95%CI:1.34～2.47)更易引发再生障碍性贫血。结论 有机磷农药可能是再生障碍性贫血的危险因素。在日常生活和工农业生产中,减少有机磷农药的暴露,可以预防再生障碍性贫血的发生。     

Estrogenic and antiestrogenic activities of flavonoid phytochemicals through estrogen receptor binding-dependent and -independent mechanisms

Members of the flavonoid class of phytochemicals have previously been demonstrated to possess estrogenic activity in a number of hormonally responsive systems. We have performed the present study to characterize the estrogenic and antiestrogenic activity of flavonoids in the estrogen receptor (ER)-positive MCF-7 human breast cancer cell line. Using an ER-dependent reporter gene assay and an ER competition binding assay, we have identified phytochemicals possessing estrogenic and antiestrogenic activities, which appeared to correlate directly with their capacity to displace [3H]estradiol from ER. Several flavonoids, including kaempferide, apigenin, and flavone, were distinct, in that their antiestrogenic activity did not appear to correlate with binding to ER, and therefore their suppression of estrogen-mediated gene transactivation and proliferation may occur independent of direct antagonism of the receptor. Further examination in HEK-293 cells transfected with ERalpha or ERbeta demonstrated potent antagonism with kaempferide and apigenin, while flavone was weakly antagonistic only toward ERP. These results suggest that the receptor binding-independent antiestrogenic chemicals may function through alternate signaling pathways as indirect ER modulators in a receptor- and cell type-specific manner. We conclude that antiestrogenic activities of flavonoid phytochemicals may occur through ER binding-dependent and -independent mechanisms and that the binding-independent antiestrogen activity of certain flavonoids is biologically significant in regulation of breast cancer cell proliferation.