摘要目的 通过转染GFP-C1-ERα质粒建立稳定表达ERα的MDA-MB-23l细胞株,观察ERα对该细胞株白细胞介素(IL)-8表达的影响.方法 pEGFP-C1-ERα质粒经酶切和测序后转染MDA-MB-231细胞,使用G418筛选出稳定表达的克隆并鉴定.使用荧光逆转录-聚合酶链反应(RT-PCR)分别测定稳定转染ERα的MDA-MB-231细胞、MDA-MB-231细胞及MCF-7细胞的IL-8mRNA的表达,使用酶联免疫吸附试验(ELISA)法测定细胞上清液IL-8的浓度.结果 成功建立ERα阳性表达的MDA-MB-231细胞株,转染ERα的细胞株IL-8 mRNA的合成(105±16)ng/L明显低于MDA-MB-231细胞(195±32)ng/L(P<0.05),但仍然高于MCF-7细胞(32±4)ng/L(P<0.05),转染后细胞上清液IL-8浓度较未转染细胞明显降低,分别为(3499±312)ng/L和(6788±427)ng/L(P<0.05),但仍然高于MCF-7细胞(1846±44)ng/L(P<0.05).结论 稳定转染ERα基因抑制了MDA-MB-231细胞的IL-8的合成和分泌.

abstractsObjective To construct stable MDA-MB-231 breast cancer cell line expressing estrogen receptor-α (ER-α) ,and explore the effect of ERα on interleukin (IL) -8 expression. Methods The GFP-C1-ERα plasmids were stably transfected into ER-negative MDA-MB-231 cells. Forty-eight hours posttransfection, the media was replaced with C418-containing media. Individual colonies were picked following 2 weeks of selection. The expression of IL-8 mRNA and the IL-8 secretion concentration were assayed in MDA-MB-231 ,ER-α transfected MDA-MB-231 and MCF-7 cell lines. Results The ERα stable transfectants in MDA-MB-231 cells were constructed successfully according Western blotting of ERα protein. The level of IL-8 mRNA and IL-8 secretion in ERα tranfected MDA-MB-231 cells were lower than which in MDA-MB-231 cells [(105 ±16) vs (195 ±32) ng/L,and (3499 ±312) vs (6788 ±427) ng/L,P <0. 05]but were still higher than which in MCF-7 cells [(32 ± 4) ng/L and (1846 ± 44) ng/L, P <0. 05]. Conclusion ERα stable transfection inhibits the expression and secretion of IL-8.