Reversal of P-glycoprotein-mediated multidrug resistance by CD44 antibody-targeted nanocomplexes for short hairpin RNA-encoding plasmid DNA delivery
Reversal of P-glycoprotein-mediated multidrug resistance by CD44 antibody-targeted nanocomplexes for short hairpin RNA-encoding plasmid DNA delivery
复制标题
通过 CD44 抗体靶向纳米复合物逆转 P-糖蛋白介导的多药耐药性，用于短发夹 RNA 编码质粒 DNA 递送。
DOI：
10.1016/j.biomaterials.2014.12.030
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发表时间：
2015-03-01
期刊：
影响因子：
14
通讯作者：
Sha, Xianyi
中科院分区：
文献类型：
--
作者：
Gu, Jijin;Fang, Xiaoling;Sha, Xianyi
多药耐药(MDR)仍然是许多化疗药物在癌症治疗中疗效降低的主要原因之一。三磷酸腺苷结合盒(ABC)转运蛋白P-糖蛋白(MDR1/P-gp)是一种具有异常活性的外排蛋白，与肿瘤的多药耐药有关。为利用RNA干扰(RNAi)技术逆转MDR，构建了编码抗MDR11P-gp短发夹状RNA(ShRNA)分子的U6-RNA基因启动子驱动的表达载体(简称PDNA-iMDR1-shRNA)。本研究探讨以Pluronic P123-偶联的聚丙基亚胺(PPI)树枝状大分子(P123-PPI)作为PDNA-iMDR1-shRNA载体的可行性，以克服乳腺癌细胞对肿瘤的耐药性。抗CD44单抗(CD44受体靶向配体)修饰的P123-PPI(抗CD44-P123-PPI)可以有效地将PDNA凝聚成纳米复合体，从而实现PDNA的高效传递、肿瘤特异性和长循环。体外实验系统地评价了P123-PPI和抗CD44-P123-PPI对PDNA-iMDR1-shRNA转运和P-gp下调的影响。体外实验结果表明，P123-PPI/PDNA和抗CD44-P123-PPI/PDNA纳米复合体具有较低的细胞毒性，与PPI/PDNA纳米复合体和脂质体(TM)2000相比，在有血清存在的情况下具有更高的转染率。负载抗P-gp的pDNA-iMDR1-shRNA的纳米复合体可以在mRNA和蛋白水平逆转MDR，同时抑制MDR1/P-gp的表达，并改善阿霉素(ADR)在MCF-7/ADR多药耐药细胞系中的内化和细胞毒作用。以荷MCF-7/ADR的BALB/c裸鼠为移植瘤模型，评价其体内抗肿瘤作用。结果表明，抗CD44-P123-PPI/PDNA-iMDR1-shRNA纳米复合体与ADR联合应用能更有效地抑制肿瘤生长。ADR的疗效增强可能与ADR在耐药肿瘤细胞内蓄积增加有关。因此，这些结果表明，使用pDNA-iMDR1-shRNA纳米复合体可能是逆转MDR和提高化疗效果的一种有前途的基因输送策略。(C)2014爱思唯尔有限公司。保留所有权利。
Multidrug resistance (MDR) remains one of the major reasons for the reductions in efficacy of many chemotherapeutic agents in cancer therapy. As a classical MDR phenotype of human malignancies, the adenosine triphosphate binding cassette (ABC)-transporter P-glycoprotein (MDR1/P-gp) is an efflux protein with aberrant activity that has been linked to multidrug resistance in cancer. For the reversal of MDR by RNA interference (RNAi) technology, an U6-RNA gene promoter-driven expression vector encoding anti-MDR11P-gp short hairpin RNA (shRNA) molecules was constructed (abbreviated pDNA-iMDR1-shRNA). This study explored the feasibility of using Pluronic P123-conjugated polypropylenimine (PPI) dendrimer (P123-PPI) as a carrier for pDNA-iMDR1-shRNA to overcome tumor drug resistance in breast cancer cells. P123-PPI functionalized with anti-CD44 monoclonal antibody (CD44 receptor targeting ligand) (anti-CD44-P123-PPI) can efficiently condense pDNA into nanocomplexes to achieve efficient delivery of pDNA, tumor specificity and long circulation. The in vitro studies methodically evaluated the effect of P123-PPI and anti-CD44-P123-PPI on pDNA-iMDR1-shRNA delivery and P-gp downregulation. Our in vitro results indicated that the P123-PPI/pDNA and anti-CD44-P123-PPI/pDNA nanocomplexes with low cytotoxicity revealed higher transfection efficiency compared with the PPI/pDNA nanocomplexes and Lipofectamine (TM) 2000 in the presence of serum. The nanocomplexes loaded with pDNA-iMDR1-shRNA against P-gp could reverse MDR accompanied by the suppression of MDR1/P-gp expression at the mRNA and protein levels and improve the internalization and cytotoxicity of Adriamycin (ADR) in the MCF-7/ADR multidrug-resistant cell line. BALB/c nude mice bearing MCF-7/ADR tumor were utilized as a xenograft model to assess antitumor efficacy in vivo. The results demonstrated that the administration of anti-CD44-P123-PPI/pDNA-iMDR1-shRNA nanocomplexes combined with ADR could inhibit tumor growth more efficiently than ADR alone. The enhanced therapeutic efficacy of ADR may be correlated with increased accumulation of ADR in drug-resistant tumor cells. Consequently, these results suggested that the use of pDNA-iMDR1-shRNA-loaded nanocomplexes may be a promising gene delivery strategy to reverse MDR and improve the effectiveness of chemotherapy. (C) 2014 Elsevier Ltd. All rights reserved.