摘要
目的筛选有效的MRP1 siRNA序列,为RNAi的体内外研究提供依据。方法利用Dharmacon公司的RNA在线工具,设计了4对针对MRP1基因不同区域的siRNA序列,分别转染至U251等三种肿瘤细胞,采用荧光转染试剂观察转染效率;采用RT-PCR和Western blot检测MRP1 mRNA和蛋白的表达。结果 siRNAl、siRNA2、siRNA3、siRNA4四对序列分别转染上述三种肿瘤细胞后的RT-PCR实验结果说明siRNA2和siRNA4对MRP mRNA有较强的抑制作用。结论 siRNA2、siRNA4是可有效抑制MRP基因表达的序列。
Objective Selecting transfected siRNAs and try to find a novel way to overcome multidrug resistance （MDR） in cancer therapy. Methods Four different MRPl-targeted siRNA duplexes were designed and synthesized. In vitro, three tumor cell lines in exponential phase of growth were transfected with the four siRNA duplexes as directed by the manufacturer protocol for lipofectamine TM2000-based transfection. Silencing was examined 24 and 48 hours after transfeetion by reverse transcription-PCR （RT-PCR）. Results After transfected to three tumor cell lines, the results of RT-PCR showed that the four siRNA duplexes significantly reduced the levels of the MRP1 mRNA and siRNA2 and siRNA4 had the better silencing effect. Conclusions Our research showed that transfection of siRNA targeted MRP1 gene can effectively inhibit the mRNA of MRP.
出处
《临床神经外科杂志》
CAS
2011年第6期281-284,共4页
Journal of Clinical Neurosurgery
基金
2005年南京市卫生科技发展重点项目(ZKX05002)
2006年南京市卫生科技发展重点项目(ZKM06056)
