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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 ERRα in estrogen receptor (ER) negative and positive endometrial cancer cells. Methods Eukaryotic expression plasmid pSG-ERRα (0.5, 1.0, 1.5 and 2.5 μg) was transiently transfected into endometrial carcinoma cell line HEC-1A (ER negative), HEC-1B (ER negative), Ishikawa (ER positive) The expression of ERRα mRNA and protein was detected by quantitative RT-PCR and western blot. The cell cycle was analyzed by flow cytometry, and the proliferation of cells was counted. Results After transfection with pSG-ERRα plasmid, the expression of ERRα was detected at mRNA and protein levels in HEC-1A, HEC-1B and Ishikawa cells. The expression levels of ERRα mRNA at the end of transfection of HEC-1B, HEC-1A and Ishikawa cells were 2104.2, 2870.6 and 1476.8 copies / ng, respectively. The expression levels of ERRα mRNA were 9835.3, 9644.4 and 8008.6 copies / , Respectively, compared with their respective untransfected cells, the differences were statistically significant (P values were 0.004,0.002,0.002). The expression levels of ERRα protein in HEC-1A, HEC-1B and Ishikawa cells were 0.823, 0.192 and 0.673, respectively. The expression levels of ERRα protein were 1.128, 1.104 and 1.008 respectively after transfection, Dyed cells, the differences were statistically significant (P <0.05). With the increase of plasmid pSG-ERRα, the percentage of S phase and G 2 / M phase of HEC-1A and HEC-1B cells increased significantly (P <0.01). After transfection with 0.5 and 1.0 μg pSG-ERRα plasmid, the growth rate of HEC-1B and HEC-1A cells was significantly increased from 24 h to 96 h after transfection (P <0.05). Conclusion ERRα overexpression is a cell proliferation mechanism of ER-negative endometrial carcinoma cell lines HEC-1A and HEC-1B.