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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 To screen effective MRP1 siRNA sequences and provide basis for RNAi in vitro and in vivo. Methods Four siRNA sequences against different regions of MRP1 gene were designed by Dharmacon RNA online tool and transfected into three kinds of tumor cells such as U251 respectively. The transfection efficiency was observed by fluorescent transfection reagent. The transfection efficiency was analyzed by RT-PCR and Western blot Detection of MRP1 mRNA and protein expression. Results RT-PCR results showed that siRNA2 and siRNA4 had a strong inhibitory effect on MRP mRNA after transfection of the three tumor cell lines with siRNA4, siRNA2, siRNA3 and siRNA4 respectively. Conclusion siRNA2 and siRNA4 are sequences that can effectively inhibit MRP gene expression.