乳腺癌耐药细胞ER表达状态影响细胞对屈洛昔芬和阿霉素的敏感性  

Status of Estrogen Receptor Affects the Drug Sensitivity of Drug Resistant MCF-7/Adr Human Breast Cancer Cells to Droloxifene and Adriamycin

作　　者：高海东[1] 孙靖中[1] 毕冬松[1] 马榕[1] 

机构地区：[1]山东大学齐鲁医院普通外科,山东济南250012

出　　处：《癌症》2003年第4期376-379,共4页Chinese Journal of Cancer

摘　　要：背景与目的:雌激素受体(estrogenreceptor,ER)阳性乳腺癌细胞株来源的耐药细胞株中,ER表达缺失或下降,且细胞生长速度减慢。本文的目的是研究MCF-7/Adr乳腺癌耐药细胞株中ER表达状态与细胞对屈洛昔芬(droloxifene,Dro)和阿霉素(Adriamycin,Adr)敏感性之间的关系。方法:Westernblot法检测MCF-7/Adr及其亲本MCF-7细胞中ER蛋白的表达,构建ER的真核细胞表达质粒(pCER),利用LipofectAMINETM将ER基因导入MCF-7/Adr细胞,经G418抗性筛选获得阳性克隆(MTER/Adr),PCR、Westernblot法鉴定并检测ER基因的整合和蛋白表达,流式细胞仪检测细胞周期变化,MTT法检测Dro及Adr对细胞增殖的影响。结果:在MCF-7细胞中可检测到ER蛋白的表达,而MCF-7/Adr细胞中使用Westernblot检测不到ER蛋白的表达。成功构建真核细胞表达质粒pCER并转染MCF-7/Adr细胞,阳性克隆MTER/Adr整合了ER基因并获得表达。经MTT分析,10μmol/LDro对MCF-7细胞的生长有明显的抑制作用,而到20μmol/L时对MCF-7/Adr细胞的生长有抑制作用。ER转染MCF-7/Adr细胞后,15μmol/L的Dro对其生长出现抑制作用,使细胞多分布于G0/G1期。同时细胞对Adr的敏感性下降。结论:MCF-7/Adr细胞中ER的丢失引起细胞对以ER为靶点的Dro治疗不敏感。BACKGROUND ＆OBJECTIVE: Loss or decreased expression of estrogen receptor (ER) and decreased growth rate regularly occur in drug resistant breast cancer cells. This study was designed to investigate the effect of estrogen receptor status on the drug resistance to droloxifene (Dro) and Adriamycin (Adr) of drug resistant MCF 7/Adr human breast cancer cells. METHODS: The expression of ER in MCF 7 and MCF 7/Adr cells was determined using Western blot analysis. ER expression plasmid was constructed and introduced into MCF 7/Adr cells using LipofectAMINETM. After G418 screening, the positive clone (MTER/Adr) was obtained. The integration and expression of ER gene were analyzed by polymerase chain reaction (PCR) and Western blot. The cell cycle distribution was investigated by flow cytometry. The effects of droloxifene and Adriamycin on the growth of cells were investigated by MTT assay. RESULTS: Western blot analysis showed that ER was positive in MCF 7 cells, but was negative in MCF 7/Adr cells. The ER expression plasmid was constructed and introduced into MCF 7/Adr cells. The integration and expression of ER gene were successful in positive clone MTER/Adr cells. Droloxifene inhibited the growth of MCF 7 at the concentration of 10 20 μmol/L and the MCF 7/Adr only at concentration of 20 μmol/L. Droloxifene inhibited the growth of MTER/Adr at the concentration of 15 μmol/L, and the percentage of MTER/Adr cells increased in G0/G1 phase. The sensitivity of MTER/Adr cells to Adriamycin increased. CONCLUSION: The insensitivity of MCF 7/Adr human breast cancer cells to droloxifene was associated with the loss of ER. MTER/Adr cells partially restore the sensitivity to droloxifene and Adriamycin.

关 键 词：乳腺肿瘤 雌激素受体 耐药性 内分泌治疗 

分 类 号：R737.9[医药卫生—肿瘤] R730.54[医药卫生—临床医学]