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http://dx.doi.org/10.7314/APJCP.2014.15.24.10597

RNAi-based Knockdown of Multidrug Resistance-associated Protein 1 is Sufficient to Reverse Multidrug Resistance of Human Lung Cells  

Shao, Shu-Li (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Cui, Ting-Ting (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Zhao, Wei (Beijing systerm for Diseases Control and Prevention)
Zhang, Wei-Wei (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Xie, Zhen-Li (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Wang, Chang-He (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Jia, Hong-Shuang (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Liu, Qian (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.15, no.24, 2015 , pp. 10597-10601 More about this Journal
Abstract
Up-regulation of multidrug resistance-associated protein 1 (MRP1) is regarded as one of the main causes for multidrug resistance (MDR) of tumor cells, leading to failure of chemotherapy-based treatment for a multitude of cancers. However, whether silencing the overexpressed MRP1 is sufficient to reverse MDR has yet to be validated. This study demonstrated that RNAi-based knockdown of MRP1 reversed the increased efflux ability and MDR efficiently. Two different short haipin RNAs (shRNAs) targeting MRP1 were designed and inserted into pSilence-2.1-neo. The shRNA recombinant plasmids were transfected into cis-dichlorodiamineplatinum-resistant A549 lung (A549/DDP) cells, and then shRNA expressing cell clones were collected and maintained. Real time PCR and immunofluorescence staining for MRP1 revealed a high silent efficiency of these two shRNAs. Functionally, shRNA-expressing cells showed increased rhodamine 123 retention in A549/DDP cells, indicating reduced efflux ability of tumor cells in the absence of MRP1. Consistently, MRP1-silent cells exhibited decreased resistance to 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DDP, suggesting reversal of MDR in these tumor cells. Specifically, MRP1 knockdown increased the DDP-induced apoptosis of A549/DDP cells by increased trapping of their cell cycling in the G2 stage. Taken together, this study demonstrated that RNAi-based silencing of MRP1 is sufficient to reverse MDR in tumor cells, shedding light on possible novel clinical treatment of cancers.
Keywords
Multidrug resistance; MRP1; ShRNA; protein expression; tumor cells;
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