| | | 农药咪鲜安雌激素样作用机制体外研究
| | 免费小论文【关键词】 咪鲜安；雌激素样作用；MCF-7细胞；雌激素受体
摘要： 目的免费小论文 用体外试验检测农药咪鲜安的免费小论文雌激素样作用，并对其可能的作用机制进行探讨。方法 将人乳腺癌细胞株（MCF-7）细胞在含10%小牛血清的RPM11640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RPMI 1640培养基中培养。设溶剂对照组、阳性对照组及5个不同浓度的咪鲜安样品组，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咪鲜安对MCF-7细胞增殖的影响，并通过雌激素受体（ER）结合试验分析咪鲜安与ER的结合情况，探讨可能的作用机制。结果 咪鲜安在10-7～10-5mol/L剂量范围内可诱导MCF-7细胞增殖，差异有统计学意义（P<005或P<001），且存在剂量-效应关系；咪鲜安能竞争性地与ER结合，从而抑制125I-E2与ER结合。结论 咪鲜安具有雌激素样作用，其作用机制在于其竞争性地与ER的结合。
关键词：咪鲜安；雌激素样作用；MCF-7细胞；雌激素受体
Study on estrogenic activity and mechanism of prochloraz in vitro
Abstrat:Objective To examine the estrogenic-like activity of the pesticide prochloraz by tests in vitro and study the possible mechanism.Methods MCF-7 cells were maintained in RPMI1640 medium containing 10% bovine serum.Two weeks before the addition of the test compound the cells were transformed into phenol red-free RPMI1640 medium containing 10% charcoal/dextran treated bovine serum.There are seven cell groups deal with different treatments:a vehicle control group,a positive group and five proliferation test,MTT test and growth curve were used to assess the effect on the proliferation of MCF-7 cells by prochloraz,and estrogen receptor(ER)binding test was used to analyze the binding ability of prochloraz with ER. Results Prochloraz can induce the proliferation of MCF-7 cells at 10-7～10-5mol/L with dose-effect relationship,and this effect is significantly different from vehicle control(P<0.05 or P<0.01).Our study also show that prochloraz can bind with ER and inhibit the bind of125I-E2 with ER.Conclusions Prochloraz has estrogenic activity which caused by its bind with ER.
Key words: prochloraz;estrogenic activity;MCF-7 cell;estrogen receptor
农药咪鲜安(prochloraz)，系咪唑类广谱杀菌剂，Vinggaard等曾报道过咪鲜安的抗雄激素活性〔1〕。本研究曾通过体内试验证明咪鲜安具有雌激素样作用〔2〕。本文在体外实验条件下观察咪鲜安对人乳腺癌细胞株MCF-7细胞增殖的影响及其与雌激素受体(ER)竞争性结合的能力，对其雌激素作用进行探讨，并研究其可能的作用机制。
1 材料与方法
11 细胞培养 人乳腺癌MCF-7细胞在本实验室常规传代培养。
12 受试物与试剂 咪鲜安原药(纯度98%，浙江绍兴东湖生化有限公司)；17-β-雌二醇(E2)(美国Sigma公司)；125I-E2及分离试剂(北京北方生物制品研究所)；MTT(上海华舜生物工程有限公司)；二甲基亚矾(DMSO，天津市化学试剂一厂)；RPMI1640培养基、无酚红RPMI1640培养基产品(美国Hyclone公司)；高融合型细胞培养用小牛血清、去激素小牛血清(上海纬群生物工程有限公司)。
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