Snail增强乳腺癌细胞MCF-7中P-gp介导的多药耐药

目的探讨乳腺癌细胞中Snail过表达与P-gp之间的关系,揭示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pCDNA3.1-Snail,将载体转染人乳腺癌细胞MCF-7后用阿霉素对细胞进行诱导。利用细胞毒性实验(MTT)、阿霉素外排实验对耐药细胞系的耐药状况进行评价;通过流式细胞术和Real-time PCR分别测量耐药细胞系中P-gp和MDR1 mRNA,Snail mRNA的表达。结果由细胞毒性实验和阿霉素外排实验的结果显示,MCF-7/Snail细胞经阿霉素诱导后相对耐药指数升高至109.2,细胞内荧光强度降至7.1(P<0.05);Real-time PCR显示相对于MCF-7,MCF-7/Snail细胞中的MDR1 mRNA,Snail mRNA的表达明显升高与流式细胞术显示的P-gp升高相一致。结论转染Snail真核表达载体后,MCF-7/Snail细胞的耐药性较MCF-7细胞明显升高。

Highexpression of Snail leads to the P-gp modulateds MDR in breast cancer cell MCF-7

Aim To explore the relationship between Snail and P-gp in breast cancer cell,and to reveal the effect of epithelial-mesenchymal transformation(EMT)on the multidrug-resistance(MDR)of breast cancer cell.Methods The eukaryotic expression vector pCDNA3.1-Snail was constructed, and then transfected into human breast cancer cell line MCF-7 to constuct MCF-7/Snail.Both cell lines MCF-7 and MCF-7/Snail were induced by adriamycin(ADM).Cell cytotoxicity assay and ADM efflux assay were used to measure the ability of drug resistance.The positive rate of P-gp of the two cell lines was detected by flow cytometry;the mRNA of MDR1 and Snail was evaluated by real-time PCR.Results MCF-7,the expression of MDR1 mRNA and Snail mRNA in MCF-7/Snail cell lines significantly increased;the expression of P-gp was increased too;the RR increased to 109.2;fluorescence intensity intracellular was reduced to 7.1.Conclusion After transfected the eukaryotic expression vector,the capacity of MCF-7/Snail strongly increases compared with that of MCF-7.