[GGI作为基因载体向A549/DDP细胞递送MDR1 siRNA以逆转多药耐药]。

药学学报 Pub Date : 2017-02-01
Zi-rui Wang, Feng Bai, Xiao-ying Zhang, Jia-min Wu, Ling Guo, Zhi Li, Min Feng
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引用次数: 0

摘要

本研究旨在通过RNA干扰(RNAi)技术下调MDR1基因,逆转肺癌细胞的多药耐药。合成了一种新型的可生物降解阳离子聚合物(PEG- b- PLG-g-PEIs,简称GGI),并用1H NMR对其进行了表征。采用动态光散射法(DLS)测定了样品的粒径和zeta电位。采用MTT法检测A549和A549/DDP细胞系GGI的细胞活力。利用流式细胞术(FCM)研究了GGI聚合物向细胞递送siRNA的效率和强度。GGI/MDR1 siRNA转染后,采用RT-PCR和Western blot检测mRNA和P-gp的表达。采用MTT法和Annexin V-FITC/PI法检测转染GGI/MDR1 siRNA复合物后顺铂给药敏感性。结果表明,GGI/siRNA的粒径为150 ~ 200 nm, zeta电位为16 ~ 28 mV。GGI具有比PEI 25K更低的细胞毒性和更高的siRNA递送效率,显著降低A549/DDP细胞MDR1 mRNA和P-gp的表达,增加A549/DDP细胞对顺铂的敏感性。GGI作为一种新型阳离子聚合物,在基因传递方面具有很大的潜力,值得进一步研究。
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[GGI as a gene carrier delivering MDR1 siRNA to A549/DDP cells for reversal of multidrug resistance].

This study was designed to reverse multidrug resistance of lung cancer cells by downregulating MDR1 genes through RNA interference (RNAi) technology. A novel biodegradable cationic polymer (PEG- b- PLG-g-PEIs, GGI) was synthesized and characterized by 1H NMR. The particle size and zeta potential were measured by dynamic light scattering (DLS). The cell viability profile of GGI was tested by MTT method with both A549 and A549/DDP cell lines. Flow cytometry (FCM) technology was used to investigate the efficiency and intensity of delivering siRNA to cells by GGI polymer. RT-PCR and Western blot were used to detect the mRNA and P-gp expression after GGI/MDR1 siRNA transfection assay. The sensitivity of cisplatin administration after transfecting GGI/MDR1 siRNA polyplexs was performed with MTT and Annexin V-FITC/PI methods. The results suggest that the particle size and zeta potential of GGI/siRNA were 150 −200 nm and 16−28 mV. GGI exhibited a lower cell cytotoxity than PEI 25K and higher efficiency of delivering siRNA, which dramatically decreased the expression of MDR1 mRNA and P-gp of A549/DDP cells and increased much sensitivity to cisplatin in A549/DDP cells. GGI holds a great potential in gene delivery as a novel cationic polymer for further investigation.

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来源期刊
药学学报
药学学报 Pharmacology, Toxicology and Pharmaceutics-Pharmacology, Toxicology and Pharmaceutics (all)
CiteScore
1.20
自引率
0.00%
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0
期刊介绍: Acta Pharmaceutica Sinica B (APSB) is a bimonthly English peer-reviewed online journal in ScienceDirect, which publishes significant original research articles, communications and high quality reviews of recent advances. APSB encourages submissions from all areas of pharmaceutical sciences, including pharmacology, pharmaceutics, medicinal chemistry, natural products, pharmacognosy, pharmaceutical analysis and pharmacokinetics. APSB is a part of the series Acta Pharmaceutica Sinica, which was founded in 1953. The journal is co-published by Elsevier B.V., in association with the Institute of MateriaMedica, Chinese Academy of Medical Sciences and Chinese Pharmaceutical Association.
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