siRNA沉默雌激素受体β表达和雌二醇干预对A549细胞的影响  

Effect of silencing estrogen receptorβby siRNA and E_2 intervention on A549 cells

作　　者：朱恪岩 古兆森[1] 闫明[2] 郭茂峰[3] 王淑玲[3] ZHU Keyan;GU Zhaosen;YAN Ming(Department of Integrated Traditional and Western Medicine,First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,CHINA)

机构地区：[1]郑州大学第一附属医院中西医结合科,河南450052 [2]河南省肿瘤医院胸外科 [3]郑州大学基础医学院

出　　处：《江苏医药》2018年第11期1225-1230,共6页Jiangsu Medical Journal

基　　金：国家自然科学基金(81473497);河南省科技厅社发攻关项目(132102310106);河南省教育厅项目(12A320077)

摘　　要：目的探讨小干扰RNA(siRNA)沉默雌激素受体β(ERβ)表达和雌二醇(E2)干预对非小细胞肺癌细胞系A549的作用及其可能分子机制。方法siRNA转染实验分为空白对照组(Control组)、siRNA-NC转染组(NC组)、Lipofectamine 2000转染组(Lipo组)和siRNA-ERβ转染组(siRNA组);E2干预实验分为0μmol/L组、10μmol/L组、50μmol/L组和100μmol/L组。采用Western blot和RT-PCR分别检测A549细胞中ERβ、表皮生长因子受体(EGFR)、磷脂酰肌醇3-激酶(PI3K)、Akt的蛋白和mRNA表达,CCK-8法和划痕实验分别检测A549细胞活性和迁移能力的变化。结果与Control组、NC组和Lipo组相比,siRNA组ERβ、EGFR、PI3K、Akt蛋白和mRNA表达下调,细胞活性和迁移能力降低(P<0.05)。与0μmol/L组相比,10μmol/L组ERβ、EGFR、PI3K、Akt蛋白和mRNA表达上调,细胞活性和迁移能力增加(P<0.05);而50μmol/L组ERβ、EGFR、PI3K、Akt蛋白和mRNA表达下调,细胞活性和迁移能力降低(P<0.05);100μmol/L组各指标均无明显变化(P>0.05)。结论 E2及其受体ERβ可能通过EGFR/PI3K/Akt信号通路影响A549细胞的活性及迁移能力,从而参与非小细胞肺癌的发生和发展过程。Objective To investigate the effect and underlying molecular mechanism of silencing estrogen receptorβ(ERβ)by small interfering RNA(siRNA)and estradiol(E2)intervention on nonsmall cell lung cancer cell line A549.Methods A549 cells were transfected with siRNA-NC(group NC),Lipofectamine 2000(group Lipo)and siRNA-ERβ(group siRNA),which without intervention were taken as group Control.Besides,A549 cells were incubated with different concentrations of E2 0,10,50 and 100μmol/L.The protein and mRNA expressions of ERβ,EGFR,PI3 Kand Akt were detected by Western blot and RT-PCR,respectively.The cell activity and migration were determined by CCK-8 and scratch assays.Results The protein and mRNA expressions of ERβ,EGFR,PI3 Kand Akt and the activity and migration of A549 cells were significantly lower than those in groups of Control,NC and Lipo(P<0.05).Compared with group 0μmol/L,the protein and mRNA expressions of ERβ,EGFR,PI3 Kand Akt and the activity and migration of A549 cells were all increased in group10μmol/L(P<0.05),which were decreased in group 50μmol/L(P<0.05)and not obviously changed in group 100μmol/L(P>0.05).Conclusion E2 and its receptor ERβmay affect the activity and migration of A549 cells through EGFR/PI3 K/Akt signaling pathway,and participate in the development and progress of non-small cell lung cancer.

关 键 词：非小细胞肺癌 雌二醇 雌激素受体Β 

分 类 号：R734[医药卫生—肿瘤]