靶向肿瘤多药耐药基因MDR1 siRNA的设计与筛选  

Design and Screening of siRNA Targeting to Multi-drug Resistance Gene MDR1

作　　者：问天娇 刘东璐[2] 田勤 崔京霞[1] 

机构地区：[1]河北医科大学药学院,河北石家庄050017 [2]河北医科大学第四医院,河北石家庄050017 [3]石家庄人民医学高等专科学校,河北石家庄050017

出　　处：《中国医药工业杂志》2016年第2期158-162,共5页Chinese Journal of Pharmaceuticals

基　　金：河北省应用基础研究计划重点基础研究项目(12966417D);河北省自然科学基金(H2012206039)

摘　　要：本研究主要考察siRNA对于逆转P-糖蛋白介导的肿瘤多药耐药的作用。以MDR1基因为靶标,在线设计MDR1基因的siRNA序列并利用生物学软件进行优化,经化学合成后转染入体外培养的乳腺癌耐药细胞系MCF-7/ADR中,经实时定量荧光PCR和Western blot定量分析MDR1基因的表达,以MTT法检测转染细胞对多柔比星的敏感性。最终设计并合成了8条MDR1基因siRNA序列,其中4条能有效抑制MDR1基因mRNA的表达。siRNA1沉默效率最高,可特异性地沉默MDR1基因,有效逆转乳腺癌细胞的多药耐药性。The effect of siRNA on the reversal of multi-drug resistance of cancer cells which was mediated by P-glycoprotein was investigated. The human MDR1 gene was selected as the target. si RNA candidate sequences were obtained via online design and further optimized by biological software. The chemically synthesized si RNAs were transfected into drug-resistant breast cancer cell line MCF-7/ADR. MDR1 gene expression were quantitatively analyzed by real-time PCR and Western blot, respectively. MTT experiment was carried out to assay the sensitivity of transfected cells to doxorubicin. Ultimately, eight MDR1 gene si RNA sequences were designed and synthesized, of which four sequences could effectively inhibit the expression of MDR1 gene. The si RNA1 had the highest interference efficiency and could specifically silence MDR1 gene, effectively reversing the multi-drug resistance in breast cancer.

关 键 词：小干扰RNA P-糖蛋白 多药耐药 乳腺癌 基因沉默 

分 类 号：Q78[生物学—分子生物学]