ERRα过度表达对子宫内膜癌Ishikawa细胞内分泌治疗的影响

Over expression of estrogen receptor-related receptor α provide a potential resistant mechanism to endocrine therapy

目的探讨孤儿受体-雌激素受体相关受体α(ERRα)在子宫内膜癌Ishikawa细胞中过度表达对内分泌治疗的影响。方法重组并构建含有绿色荧光蛋白(GFP)和G418耐药筛选标记的真核表达质粒pEGFP-N1-3FLAG-ERRα,瞬时转染子宫内膜癌细胞株Ishikawa后采用实时荧光定量PCR技术和蛋白印迹法检测ERRαmRNA和蛋白的表达情况。使用不同浓度的ICI182780处理子宫内膜癌Ishikawa细胞、Ishikawa-空载体细胞(Ishikawa-空)和ERRα过度表达的(Ishikawa-ERRα)细胞,流式细胞技术检测分析10-6mol/LICI182780对细胞诱导的凋亡情况。结果瞬时转染pEGFP-N1-ERRα质粒后,Ishikawa在mRNA和蛋白水平均能检测到ERRα的表达增加。流式细胞仪细胞凋亡分析表明,ERRα过度表达导致子宫内膜癌细胞Ishikawa耐受ICI182780的诱导凋亡作用(P<0.05)。结论 ERRα过度表达为激素依赖性的子宫内膜癌细胞株Ishikawa提供了一种耐受内分泌治疗的机制。

Objective To discuss the function of over-expression of ERRα in endometrial cancer cells and the potential role of ERRα in the resistant mechanism to endocrine therapy.Methods Plasmid pEGFP-N1-3FLAG-ERRα was reconstructed and transiently transfected into endometrial cancer cell Ishikawa.Real-time quantitative PCR and western-blot was used to analyze the mRNA and protein expression of ERRα.After treated with 10-6 mol/L ICI182780,the cellular apoptosis was analyzed in the Ishikawa cells,Ishikawa cells with empty vector（Ishikawa-vector）as well as the cells with ERRα over-expression（Ishikawa-ERRα）by using Annexin V-plus kit and flowcytomertry.Results After transfection,the cell lines were detected an increasing expression of ERRα in mRNA levels and protein levels.In endometrial cancer cells,over-expression of ERRα achieved a resistance to the cell apoptosis induced by high dose of ICI182780（10-6mol/L,pure anti-ER）.Conclusions Over-expression of ERRα may provide a molecular mechanism to resist the pure anti-ER endocrine therapy by repressing ER function.ERRα may be the next endocrine therapy target in hormone-related cancer.