有机氯类农药在残留剂量下联合诱导乳腺癌MCF-7细胞增殖的机制研究Organochlorine Pesticides Combined to Induce Proliferation of Breast Cancer MCF-7 Cells at the Residual Dose: A Mechanism Study
郎朗;董晓琪;狄静波;
摘要(Abstract):
有机氯类农药为已确定的环境雌激素,易在体内蓄积产生慢性毒性危害人体健康。选择单一存在时无明显雌激素效应的残留剂量进行联合试验,观察联合作用后雌激素效应的增减。选取乳腺癌MCF-7细胞作为研究对象,对残留剂量下六氯苯(HCB)、β-六六六(β-BHC)和p,p’-滴滴涕(p,p’-DDT)联合后的雌激素效应机制进行了研究。首先,运用MTT法观察3种有机氯类农药残留剂量下单独和联合作用对MCF-7细胞生长的影响;然后,运用流式细胞仪测定有机氯类农药联合对MCF-7细胞周期的影响;之后,采用Western Blot法测定有机氯类农药联合后ERα、ERβ、ERK1/2、p-ERK1/2、Ki67、c-Myc和Cyclin D1蛋白表达的变化。结果显示,β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组作用24 h后细胞增殖率为115.0%、120.1%,作用48 h后增殖率为121.2%、126.3%,其他联合组与对照相比未发生明显变化;β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组中MCF-7细胞的G1期细胞比例下降,S期细胞比例上升,细胞呈高增殖状态;β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组和雌二醇(E2)组作用48 h后,均降低ERα的蛋白表达,升高ERβ的蛋白表达,促进p-ERK1/2、Ki67、Cyclin D1和c-Myc的蛋白表达。研究表明,在单一作用时不具有雌激素效应的残留剂量下,仅β-BHC+p,p’-DDT联合组和HCB+β-BHC+p,p’-DDT联合组具有明显雌激素效应,其效应机制为经雌激素受体途径,激活ERβ蛋白表达,抑制ERα蛋白表达,促进p-ERK1/2、Ki67、c-Myc和Cyclin D1等相关蛋白的表达。
关键词(KeyWords): 有机氯类农药;MCF-7细胞;环境雌激素效应;联合作用;残留剂量
基金项目(Foundation): 2016年哈尔滨商业大学青年创新人才支持项目(2016QN058);; 黑龙江省教育厅科学技术研究项目(12541188);; 哈尔滨市青年项目(2013RFQXJ054);; 2017年哈尔滨商业大学研究生创新科研项目(YJSCX2017-459HSD);; 2018年哈尔滨商业大学研究生创新科研项目(YJSCX2018-549HSD);; 大学生创新创业训练计划项目(201910240025)
作者(Author): 郎朗;董晓琪;狄静波;
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