人三阴性乳腺癌细胞耐药株的建立及特性研究  

Establishment of drug-resistant cell lines of human triple negative breast cancer and study on their characteristics

作　　者：吴月琴[1] 钟山亮[2] 张晓慧[2] 唐金海[3] 赵建华[2] 

机构地区：[1]南京医科大学研究生学院,南京211166 [2]南京医科大学附属江苏省肿瘤医院临床检验中心,南京210009 [3]江苏省肿瘤医院普外科,南京210009

出　　处：《国际检验医学杂志》2016年第15期2049-2051,2054,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(81272470)

摘　　要：目的建立人三阴性乳腺癌细胞株MDA-MB-231的多西紫杉醇(Doc)耐药模型(MDA-MB-231/Doc)和表阿霉素(Epi)耐药模型(MDA-MB-231/Epi),探讨其生物学特性。方法采用Doc和EPi低浓度逐步加量诱导法历时12个月分别建立MDA-MB-231/Doc和MDA-MB-231/Epi耐药细胞株。通过细胞形态学观察、MTT法和流式细胞术分析、比较其生物学特性,实时荧光定量PCR检测多药耐药基因(MDR1)mRNA表达,Western Blot法检测P糖蛋白(P-gp)、雌激素受体(ER)、孕激素受体(PR)和人表皮生长因子受体2(Her-2)的表达状况。结果所构建的MDA-MB-231/Doc和MDA-MB-231/Epi耐药株可分别在12nmol/L Doc和800nmol/L Epi中稳定生长,在相同的药物浓度下,耐药细胞株的生长增殖率明显高于亲代细胞,其耐药指数分别为亲代敏感细胞的8.32倍和64.93倍,且相互呈交叉耐药状态。与亲代细胞相比,两株耐药细胞处于G1期和G2期的细胞增加、处于S期的细胞减少,随撤药时间的延长,细胞的增殖速度加快。两株耐药株的MDR1基因表达水平增高,分别为亲代细胞的4.05倍和5.96倍,P-gp表达为阳性。与MCF-7细胞株相比,MDA-MB-231细胞株ER、PR、HER2表达阴性,是典型的三阴性乳腺癌细胞株。结论成功建立MDA-MB-231/Doc和MDA-MB-231/Epi的耐药细胞株,其生长及耐药性稳定。Objective To establish docetaxel（Doc）resistant MDA-MB-231/Doc model and epirubicin（Epi）resistant MDAMB-231/Epi mode from triple negative breast cancer cell line MDA-MB-231 and to explore their biological characteristics.Methods The MDA-MB-231/Doc and MDA-MB-231/Epi drug-resistant cell lines were respectively established by gradually increasing Doc or Epi concentrations induction method in 12 months.The biological characteristics of the cell lines were compared by the cell morphological observation,MTT and flow cytometry;the real-time fluorescent quantitative PCR was used to detect multi-drug resistance gene（MDR1）mRNA expression;the expression of P glycoprotein（P-gp）,estrogen receptor（ER）,progesterone receptor（PR）and human epidermal growth factor receptor 2（Her-2）was detected by Western Blot.Results After the 12-month induction,the established MDA-MB-231/Doc could grow stably in the medium containing 12nmol/L Doc,and MDA-MB-231/Epi could grow stably in the medium containing 800nmol/L Epi;in the same drug concentration,the growth proliferation rate of the drug resistant cell line was significantly higher than that of the parental generation cells,their drug resistance indexes were 8.32 times and 64.93 rimes of parental generation sensitive cells,moreover which showed the mutual cross drug resistant status.Compared to the parental generation cells,the cells of stage G1 and G2in two cell lines were increased and the cells of stage S were decreased,with the prolongation of drug withdrawal time,the cell proliferation speed was accelerated.The expression level of MDR1 gene was increased in the two drug-resistant cell lines,which were 4.05 times and 5.96 times of parental generation cells respectively,P-gp protein expression was positive.Compared with the MCF-7cell line,ER,PR and Her2 expression in the MDA-MB-231 cell line was negative and typical triple negative breast cancer cell line.Conclusion The drug resistance cell lines of MDA-MB-231/Doc and MDA-MB-231/Epi are successfully established

关 键 词：乳腺癌细胞 多西紫杉醇 表阿霉素 多药耐药 

分 类 号：R737.9[医药卫生—肿瘤]