Shao Shu-Li, Sun Ying-Yu, Li Xu-Yan, Zhang Wei-Wei, Fu Bo, Yun Dong-Ze, Zuo Ming-Xue
College of Life Science, Beijing Normal University, 19 Xinjiekouwai Street, Beijing 100875, China.
Cell Biol Int. 2008 Aug;32(8):893-8. doi: 10.1016/j.cellbi.2008.03.021. Epub 2008 Apr 8.
Overexpression of P-glycoprotein (P-gp), the mdr1 gene product, confers multidrug resistance (MDR) to tumor cells and often limits the efficacy of chemotherapy. This study evaluated RNAi for specific silencing of the mdr1 gene and reversion of multidrug resistance. Three different short hairpin RNAs (shRNAs) were designed and constructed in a pSilencer 3.1-H1 neo plasmid. The shRNA recombinant plasmids were transfected into HT9 leukemia cells. The RNAi effect was evaluated by real-time PCR, Western blotting and cell cytotoxicity assay. In the cell, shRNAs can specifically down-regulate the expression of mdr1, mRNA and P-gp. Resistance against harringtonine, doxorubicin and curcumin was decreased. The study indicated that shRNA recombinant plasmids could modulate MDR in vitro.
P-糖蛋白（P-gp）即mdr1基因产物的过表达赋予肿瘤细胞多药耐药性（MDR），并常常限制化疗效果。本研究评估了RNA干扰对mdr1基因的特异性沉默作用以及多药耐药性的逆转。在pSilencer 3.1-H1 neo质粒中设计并构建了三种不同的短发夹RNA（shRNA）。将shRNA重组质粒转染至HT9白血病细胞。通过实时PCR、蛋白质免疫印迹法和细胞毒性试验评估RNA干扰效果。在细胞中，shRNA能够特异性下调mdr1、mRNA和P-gp的表达。对三尖杉酯碱、阿霉素和姜黄素的耐药性降低。该研究表明，shRNA重组质粒在体外可调节多药耐药性。
