Snail沉默和过表达对ADM诱导MDR作用的研究  

Effect of silence and highexpression of Snail on MDR induced by ADM

作　　者：王辉[1] 张义军[1] 陈伟娟[2] 王晓杰[2] 李莎莎[1] 丁保清[1] 

机构地区：[1]潍坊医学院药理学教研室,山东潍坊261041 [2]潍坊医学院病理学教研室,山东潍坊261041

出　　处：《中华肿瘤防治杂志》2010年第9期651-654,672,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省自然科学基金(Y2008C75)

摘　　要：目的:研究乳腺癌细胞MCF-7中Snail表达与多柔比星(ADM)诱导的多药耐药(MDR)的关系,揭示上皮-间质转化(EMT)对乳腺癌细胞多药耐药的影响。方法:构建Snail真核表达载体pCDNA-Snail和RNA干扰载体pSUPER-siRNA,将载体转染乳腺癌细胞MCF-7;用多柔比星对细胞进行诱导。利用细胞毒性实验和多柔比星外排实验,对细胞的耐药状况进行评价;通过流式细胞术测量耐药细胞中P-糖蛋白(P-gp)表达,Real-TimePCR测量耐药细胞中MDR1、SnailmRNA的表达。结果:细胞毒性实验和多柔比星外排实验结果显示,细胞转染pCDNA-Snail和pSUPER-siRNA后经多柔比星诱导相对耐药指数(RR)分别为111.3和13.7,细胞内药物荧光强度分别为9.30±0.97和87.40±9.13;Real-TimePCR显示相对于未转染细胞,转染pCDNA-Snail后MDR1,SnailmRNA的表达明显升高,P<0.01,P-gp表达也显著升高,P<0.01;转染pSUPER-siRNA细胞则出现相反变化(P<0.01)。结论:Snail沉默和过表达分别使乳腺癌细胞的耐药能力降低或升高。OBJECTIVE：To explore the relationship between Snail and multidrug-resistance（MDR）induced by adriamycin（ADM）in breast cancer cell lines MCF-7,and reveal the effect of epithelial-mesenchymal transformation（EMT）on MDR in breast cancer cells.METHODS：The eukaryotic expression vector pCDNA-Snail and Snail siRNA vector pSUPER-siRNA were constructed;and then transfected into breast cancer cell line,induced by ADM.Cell cytotoxicity assay and ADM exclusive assay were used to measure the ability of drug resietance.The positive rate of P-glucoprotein（P-gp）of the two cell lines was detected by flow cytometry;the mRNA of MDR1 and Snail were evaluated by Real-Time PCR.RESULTS：Compared with un-transfected cells,after transfected with pCDNA-Snail,the expression of MDR1,Snail mRNA in cell lines significantly increased（P0.01）;and the expression of P-gp increased too（P0.01）;after transfected with pSUPER-siRNA,the results were opposite（P0.01）;the relative drug resistance（RR）values were 111.3 and 13.7 respectively,and fluorescence intensity intracellular values were 9.30±0.97 and 87.40±9.13 respectively.CONCLUSION：Silence and highexpression of Snail upgrade or downgrade the ability of drug resietance of breast cancer cell respectively.

关 键 词：乳腺肿瘤 RNA 小分子干扰 RNA 转染 

分 类 号：R737.9[医药卫生—肿瘤]