摘要
目的 观察对 -壬基酚 (p- NP)对 MCF- 7人类乳腺癌细胞株雌激素受体 (ER)及 ERαm RNA表达的影响 ,探讨其雌激素样活性。方法 以 RPMI 16 4 0培养液 (含 10 %小牛血清 )对 MCF- 7细胞进行半开放式贴壁培养 ,试验前以 DMEM(不含酚红 )培养液洗涤细胞 ,以含 3% C/D胎牛血清的 DMEM(不含酚红 )培养液继续培养 3d。试验设溶剂对照、阳性对照和 p- NP 5个浓度组 ,采用免疫组织化学法和定量 RT- PCR法分别对 MCF- 7细胞 ER蛋白和 ERαm RNA表达情况进行分析。结果 1× 10 - 5m ol/L p- NP作用于 MCF- 7细胞 2 4 h下调 ER蛋白表达 ;1× 10 - 6 mol/L～ 1× 10 - 5m ol/L p- NP作用于 MCF- 7细胞 2 h和 2 4 h下调 ERαm RNA表达。p- NP各浓度组均表现出 ER蛋白和 ERαm RNA表达随 p- NP作用于 MCF- 7细胞时间的延长而进一步下降的趋势。结论 p- NP可模拟雌激素下调 MCF- 7细胞 ER蛋白和 ERαm RNA表达 。
Objective To investigate the estrogenicity of p nonylphenol (p NP) through observation of its effects on expression of estrogen receptor (ER) and ER αmRNA of MCF 7 human breast cancer cells. Methods MCF 7 cells were maintained in RPMI 1640 medium containing 10% bovine serum. Before addition of p NP, the cells were washed by phenol red free DMEM medium and then maintained in phenol red free DMEM medium containing 3% charcoal/dextran treated fetal bovine serum for 3 days. The effects of five concentrations of p NP on expression of ER protein and ER αmRNA were assayed using semi quantitative immunohistochemical method and quantitative RT PCR respectively, vehicle control and positive control were used. Results Expression of ER protein in MCF 7 cells was down regulated by 1×10 -5 mol/L p NP when cells were treated for 24 h. Expression of ER αmRNA in MCF 7 cells was down regulated by 1×10 -6 mol/L 1×10 -5 mol/L p NP when cells were treated for 2 h and 24 h. Expression of both ER protein and ER αmRNA in MCF 7 cells showed a tendency of decline when the time of exposure to different concentrations of p NP was elongated. Conclusion p NP could down regulate the expression of both ER protein and ER αmRNA in MCF 7 cells, which mimicked the effects of estradiol and thus revealed the estrogenicity of p NP.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2003年第4期641-644,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金重点课题 (批准号 3 0 0 3 0 12 0 )资助
