摘要
目的探讨雌激素受体相关受体α(ERRα)在雌激素受体(ER)阴性及阳性的子宫内膜癌细胞中的作用。方法将真核表达质粒 pSG-ERRα(0.5、1.0、1.5、2.5μg)瞬时转染子宫内膜癌细胞株 HEC-1A(ER 阴性)、HEC-1B(ER 阴性)、Ishikawa(ER 阳性),采用定量 RT-PCR 技术和蛋白印迹法(western blot)检测 ERRα mRNA 和蛋白的表达情况;采用流式细胞仪分析细胞周期,并计数细胞的增殖情况。结果转染 pSG-ERRα质粒后,HEC-1A、HEC-1B、Ishikawa 细胞在 mRNA 和蛋白水平均能检测到 ERRα的表达增加。HEC-1B、HEC-1A、Ishikawa 细胞末转染时 ERRα mRNA 的表达水平分别为2104.2、2870.6、1476.8 copies/ng,转染后3者 ERRα mRNA 的表达水平分别为9835.3、9644.4、8008.6 copies/ng,分别与各自未转染的细胞比较,差异均有统计学意义(P 值分别为0.004、0.002、0.002)。HEC-1A、HEC-1B、Ishikawa 细胞未转染时 ERRα蛋白的表达水平分别为0.823、0.192、0.673,转染后3者 ERRα蛋白的表达水平分别为1.128、1.104、1.008,分别与各自未转染的细胞比较,差异均有统计学意义(P<0.05)。随着转染 pSG-ERRα质粒质量的增加,HEC-1A、HEC-1B 细胞的 S 期和 G_2/M 期细胞比例明显上升(P<0.01)。HEC-1B、HEC-1A 细胞转染0.5、1.0μg pSG-ERRα质粒后,细胞在转染后24～96 h 间生长速度显著加快(P<0.05)。结论 ERRα过度表达是 ER 阴性的子宫内膜癌细胞株 HEC-1A、HEC-1B 的一种细胞增殖机制。
Objective To investigate the role of human estrogen receptor-related receptor （ERR） α, a submember of orphan receptors, in the tumorigenesis of endometrial cancer. Methods Plasmid of pSG-ERRα was transfected into endometrial cancer cell lines HEC-1A, HEC-1B, and Ishikawa. Real-time quantitative RT-PCR and western blot were used to analyze the mRNA and protein expression of ERRor in endometrial cancer cell. Flow cytometry was used to analyze the cellular growth. Results Expressions of the ERRor were significantly increased in the endometrial cancer cells transfected with pSG-ERRα plasmid; expression of the ERRor mRNA in HEC-1A cell was 9644.4 copies/ng, HEC-1 B： 9835.3 copies/ng, and Ishikawa： 8008. 6 copies/ng （ P 〈 0.01 ） ; expression of the ERRor protein in HEC-1A cell was 1. 128, HEC-1B ： 1. 104, and Ishikawa ： 1. 008 （ P 〈 0. 05 ）. Flow cytometry showed over-expression of ERRor was accompanied by increased HEC-1A and HEC-1B cells in S and G2/M phase （P 〈0.01 ） , while this could not be observed in the estrogen receptor （ER） positive endometrial cancer cell line Ishikawa. Furthermore, cellular growth analysis showed that over-expression of ERRor induced cell growth increase of the ER negative endometn＇al cancer cells HEC-1A and HEC-1B （P 〈 0. 05）. Conclusion Over-expression of ERRor could stimulate ER negative endometrial cancer cell proliferation independent of estrogen-ER pathway.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2007年第6期408-411,共4页
Chinese Journal of Obstetrics and Gynecology
基金
国家自然科学基金(3060666)
关键词
子宫内膜肿瘤
受体
雌激素
细胞分裂
Endometrial neoplasms
Receptor, estrogen
Cell division
