| 表达BCRP的卵巢癌耐药细胞系3AO/BCRP的建立及其生物学特征 |
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| 引用本文： | 李文通,周庚寅,宋现让,迟伟玲,任瑞美,王兴武. 表达BCRP的卵巢癌耐药细胞系3AO/BCRP的建立及其生物学特征[J]. 基础医学与临床, 2006, 26(1): 61-65 |
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| 作者姓名： | 李文通 周庚寅 宋现让 迟伟玲 任瑞美 王兴武 |
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| 作者单位： | 山东大学,医学院,病理教研室,山东,济南,250012;山东省肿瘤医院,基础研究中心,山东,济南,250117;山东大学,医学院,分子生物学教研室,山东,济南,250012 |
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| 基金项目： | 中国科学院资助项目;高等学校博士学科点专项科研项目 |
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| 摘 要： | 目的探讨乳腺癌耐药蛋白(BCRP)在卵巢癌中的作用和逆转其介导的MDR,建立表达BCRP的耐药细胞系3AO/BCRP。方法提取MCF-7细胞的总RNA,设计BCRP基因上下游引物,RT-PCR克隆出BCRP基因全长并连接到pcDNA3.1(-)上,转染3AO细胞后以G418筛选存活细胞。采用米托蒽醌外排实验、细胞毒性实验、细胞群体倍增时间、定量PCR、Western blot等方法鉴定构建的耐药细胞系。结果3AO/BCRP对米托蒽醌耐药指数增加至11.58;耐药细胞外排米托蒽醌的作用增强;细胞中BCRPmRNA的水平升高,细胞能表达一定量的BCRP。结论3AO/BCRP细胞具有BCRP的耐药表型,可作为进一步研究BCRP的生物学特性以及建立其介导的耐药逆转模型。
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| 关 键 词： | 乳腺癌耐药蛋白 多药耐药 卵巢癌 基因表达 定量PCR |
| 文章编号： | 1001-6325(2006)01-0061-05 |
| 收稿时间： | 2004-11-22 |
| 修稿时间： | 2005-01-07 |
Establisment of BCRP expressing ovary cancer cell line 3AO/BCRP and its biological profile |
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| Lii Wen-tong,ZHOU Geng-yin,SONG Xian-rang,CHI Wei-ling,REN Rui-mei,WANG Xing-wu. Establisment of BCRP expressing ovary cancer cell line 3AO/BCRP and its biological profile[J]. Basic Medical Sciences and Clinics, 2006, 26(1): 61-65 |
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| Authors: | Lii Wen-tong ZHOU Geng-yin SONG Xian-rang CHI Wei-ling REN Rui-mei WANG Xing-wu |
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| Affiliation: | 1. Department of Pathology, Medical School, Shandong University, Jinan 250012 ; 2. Department of Cancer Research Center, Shandong Tumor Hospital and Institute, Jinan 250117; 3.Department of Department of Biochemistry and Molecular Biology, Medical School, Shandong University, Jinan 250012, China |
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| Abstract: | Objective To explore the function of BCRP in ovary cancer and establisment of BCRP expressing ovary cancer cell line 3AO/BCRP.Methods We construct the BCRP expressing cell line 3AO/BCRP.Extract the total RNA of MCF-7 cells,clone the whole length of BCRP gene by RT-PCR and ligate the gene to pcDNA3.1(-),transfect the positive clones to 3AO cells and select positive cell clones with G418,identify the survival cells by MTT,efflux assay,western blot.Results The multidrug resistance index of 3AO/BCRP to mitoxantrone increased to 11.58 times;the efflux of 3AO/BCRP to mitoxantrone enhanced;the relative level of BCRP mRNA in 3AO/BCRP increased and western blot indicate that the 3AO/BCRP cells can express more protein than 3AO cells.Conclusion 3AO/BCRP is a reliable BCRP expressing cell model,the establishment of this cell line has laid basis for further study of the mechanism of BCRP. |
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| Keywords: | BCRP MDR ovary cancer gene expression real-time PCR |
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