雌激素受体β1在MDA-MB-231人乳腺癌细胞中高表达对雷诺昔芬抗肿瘤作用的影响  

Anticancer activity of raloxifene impacted by high expression of ERβ1 in MDA-MB-231 cell

作　　者：姜华[1] 李梓雄 程林 王佳妮[1] 黄勇[1] 李玺[1] 刘仁斌[1] 

机构地区：[1]中山大学附属第三医院甲状腺乳腺外科,广州510630

出　　处：《中华普通外科学文献（电子版）》2018年第1期14-18,共5页Chinese Archives of General Surgery(Electronic Edition)

基　　金：广东省医学科学技术研究基金资助项目(A2017506)

摘　　要：目的探究雷诺昔芬(RAL)对乳腺癌中过表达雌激素受体(ER)β1的MDAMB-231细胞的抑制作用。方法用含ERβ1基因的HIV慢病毒转染乳腺癌MDA-MB-231细胞,获得稳定表达ERβ1的乳腺癌细胞株。采用RT-PCR法检测转染细胞中ERβ1 m RNA的表达水平,Western blotting法检测细胞中ERβ1蛋白的表达水平;各细胞亚型分别于0、1、10μmol/L的RAL共培养48 h,观察RAL对ERβ1高表达细胞株及对照细胞株治疗作用的差异。结果 HIV慢病毒转染后细胞绿色荧光表达效率达到90%以上;RT-PCR检测显示阳性转染组MDA-MB-231/ERβ1细胞中ERβ1 m RNA水平较阴性转染组提高了12.9倍(P<0.05);阳性转染组细胞中ERβ1蛋白水平也相应提高;细胞实验表明高浓度(10μmol/L)的RAL对MDA-MB-231细胞有明显的增殖抑制作用,特别是对ERβ1高表达细胞株的抑制作用更加明显(F=9.273,P=0.015)。结论在乳腺癌MDA-MB-231细胞中,ERβ1高表达可以使细胞对RAL更加敏感。Objective To up-regulate ERβ1 m RNA in breast cancer cells,and to explore the inhibiting effect of raloxifene（RAL） on MDA-MB-231 cell which has the high expression of ERβ1.Methods We used HIV lentivirus containing ERβ1 gene to infect breast cancer cell line of MDAMB-231,and established cell-line stably over-expressing ERβ1 gene.Fluorescence microscopy was used to show the transfected cells of which the e GFP was positive.Then we identified the ERβ1 gene expression on the levels of m RNA and protein by RT-PCR and Western blotting.All cell subtypes were cultured with increasing concentration of RAL（0,1,10 μmol/L） for 48 h,the difference of therapeutic effects among the MDA-MB-231/ERβ1,MDA-MB-231/e GFP and MDA-MB-231 were detected.Results Fluorescence microscopy showed that more than 90% of transfected cells by the HIV lentivirus were green fluorescence positive.RT-PCR test showed that the levels of ERβ1 m RNA in MDA-MB-231/ERβ1 cells were increased 12.9 times respectively compared with parental cells（P〈0.05）.Western blotting showed that ERβ1 protein levels were also increased correspondingly.Cell experiment showed that high dose（10 μmol/L） of RAL inhibited proliferation of all MDA-MB-231 sub-lines,and the greater extent was observed when ERβ1 overexpressed（F=9.273,P=0.015）.Conclusion The sensitivity to RAL for breast cancer cell of MDA-MB-231 can be increased by up-regulating ERβ1 level.

关 键 词：乳腺肿瘤 雌激素受体Β 选择性雌激素受体调节剂 MDA-MB-231细胞 雷诺昔芬 

分 类 号：R737.9[医药卫生—肿瘤]