Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells

Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells

Bao-an Chen1, Pei-pei Mao1, Jian Cheng1, Feng Gao1, Guo-hua Xia1, Wen-lin Xu2, Hui-lin Shen2, Jia-hua Ding1, Chong Gao1, Qian Sun1, Wen-ji Chen1, Ning-na Chen1, Li-jie Liu3, Xiao-mao Li4, Xue-mei Wang51Department of Hematology, The Affiliated Zhongda Hospital, Clinical Medical School, Southeast Univ...

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Autores principales: Bao-an Chen, Pei-pei Mao, Jian Cheng, et al
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2010
Materias:
Medicine (General)
R5-920
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spelling oai:doaj.org-article:b28c766a8ac7409d9bfdddbbda3b64142021-12-02T02:30:50ZReversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells1176-91141178-2013https://doaj.org/article/b28c766a8ac7409d9bfdddbbda3b64142010-06-01T00:00:00Zhttp://www.dovepress.com/reversal-of-multidrug-resistance-by-magnetic-fe3o4-nanoparticle-a4629https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Bao-an Chen1, Pei-pei Mao1, Jian Cheng1, Feng Gao1, Guo-hua Xia1, Wen-lin Xu2, Hui-lin Shen2, Jia-hua Ding1, Chong Gao1, Qian Sun1, Wen-ji Chen1, Ning-na Chen1, Li-jie Liu3, Xiao-mao Li4, Xue-mei Wang51Department of Hematology, The Affiliated Zhongda Hospital, Clinical Medical School, Southeast University, Nanjing, People’s Republic of China; 2Department of Hematology, The Affiliated People’s Hospital, Jiangsu University, Zhenjiang, People’s Republic of China; 3Institution of Physiology, Southeast University, Nanjing, People’s Republic of China; 4Department of Physics, University of Saarland, Saarbruecken, Germany; 5State Key Lab of Bioelectronics (Chien-Shiung Wu Laboratory), Southeast University, Nanjing, People’s Republic of ChinaAbstract: In many instances, multidrug resistance (MDR) is mediated by increasing the expression at the cell surface of the MDR1 gene product, P-glycoprotein (P-gp), a 170-kD energy-dependent efflux pump. The aim of this study was to investigate the potential benefit of combination therapy with magnetic Fe3O4 nanoparticle [MNP (Fe3O4)] and MDR1 shRNA expression vector in K562/A02 cells. For stable reversal of “classical” MDR by short hairpin RNA (shRNA) aiming directly at the target sequence (3491–3509, 1539–1557, and 3103–3121 nucleotide) of MDR1 mRNA. PGC silencer-U6-neo-GFP-shRNA/MDR1 called PGY1–1, PGY1–2, and PGY1–3 were constructed and transfected into K562/A02 cells by lipofectamine 2000. After transfected and incubated with or without MNP (Fe3O4) for 48 hours, the transcription of MDR1 mRNA and the expression of P-gp were detected by quantitative real-time PCR and Western-blot assay respectively. Meanwhile intracellular concentration of DNR in K562/A02 cells was detected by flow cytometry (FCM). PGC silencer-U6-neo-GFP-shRNA/MDR1 was successfully constructed, which was confirmed by sequencing and PGY1–2 had the greatest MDR1 gene inhibitory ratio. Analysis of the reversal ratio of MDR, the concentration of daunorubicin (DNR) and the transcription of MDR1 gene and expression of P-gp in K562/A02 showed that combination of DNR with either MNP (Fe3O4) or PGY1–2 exerted a potent cytotoxic effect on K562/A02 cells, while combination of MNP (Fe3O4) and PGY1–2 could synergistically reverse multidrug resistance. Thus our in vitro data strongly suggested that a combination of MNP (Fe3O4) and shRNA expression vector might be a more sufficient and less toxic anti-MDR method on leukemia. Keywords: K562/A02 cell line, multidrug resistance, magnetic nanoparticle of Fe3O4, recombinant plasmid vector PGY1–2 Bao-an ChenPei-pei MaoJian Chenget alDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2010, Iss default, Pp 437-444 (2010)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Bao-an Chen
Pei-pei Mao
Jian Cheng
et al
Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
description Bao-an Chen1, Pei-pei Mao1, Jian Cheng1, Feng Gao1, Guo-hua Xia1, Wen-lin Xu2, Hui-lin Shen2, Jia-hua Ding1, Chong Gao1, Qian Sun1, Wen-ji Chen1, Ning-na Chen1, Li-jie Liu3, Xiao-mao Li4, Xue-mei Wang51Department of Hematology, The Affiliated Zhongda Hospital, Clinical Medical School, Southeast University, Nanjing, People’s Republic of China; 2Department of Hematology, The Affiliated People’s Hospital, Jiangsu University, Zhenjiang, People’s Republic of China; 3Institution of Physiology, Southeast University, Nanjing, People’s Republic of China; 4Department of Physics, University of Saarland, Saarbruecken, Germany; 5State Key Lab of Bioelectronics (Chien-Shiung Wu Laboratory), Southeast University, Nanjing, People’s Republic of ChinaAbstract: In many instances, multidrug resistance (MDR) is mediated by increasing the expression at the cell surface of the MDR1 gene product, P-glycoprotein (P-gp), a 170-kD energy-dependent efflux pump. The aim of this study was to investigate the potential benefit of combination therapy with magnetic Fe3O4 nanoparticle [MNP (Fe3O4)] and MDR1 shRNA expression vector in K562/A02 cells. For stable reversal of “classical” MDR by short hairpin RNA (shRNA) aiming directly at the target sequence (3491–3509, 1539–1557, and 3103–3121 nucleotide) of MDR1 mRNA. PGC silencer-U6-neo-GFP-shRNA/MDR1 called PGY1–1, PGY1–2, and PGY1–3 were constructed and transfected into K562/A02 cells by lipofectamine 2000. After transfected and incubated with or without MNP (Fe3O4) for 48 hours, the transcription of MDR1 mRNA and the expression of P-gp were detected by quantitative real-time PCR and Western-blot assay respectively. Meanwhile intracellular concentration of DNR in K562/A02 cells was detected by flow cytometry (FCM). PGC silencer-U6-neo-GFP-shRNA/MDR1 was successfully constructed, which was confirmed by sequencing and PGY1–2 had the greatest MDR1 gene inhibitory ratio. Analysis of the reversal ratio of MDR, the concentration of daunorubicin (DNR) and the transcription of MDR1 gene and expression of P-gp in K562/A02 showed that combination of DNR with either MNP (Fe3O4) or PGY1–2 exerted a potent cytotoxic effect on K562/A02 cells, while combination of MNP (Fe3O4) and PGY1–2 could synergistically reverse multidrug resistance. Thus our in vitro data strongly suggested that a combination of MNP (Fe3O4) and shRNA expression vector might be a more sufficient and less toxic anti-MDR method on leukemia. Keywords: K562/A02 cell line, multidrug resistance, magnetic nanoparticle of Fe3O4, recombinant plasmid vector PGY1–2
format article
author Bao-an Chen
Pei-pei Mao
Jian Cheng
et al
author_facet Bao-an Chen
Pei-pei Mao
Jian Cheng
et al
author_sort Bao-an Chen
title Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
title_short Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
title_full Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
title_fullStr Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
title_full_unstemmed Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells
title_sort reversal of multidrug resistance by magnetic fe3o4 nanoparticle copolymerizating daunorubicin and mdr1 shrna expression vector in leukemia cells
publisher Dove Medical Press
publishDate 2010
url https://doaj.org/article/b28c766a8ac7409d9bfdddbbda3b6414
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AT jiancheng reversalofmultidrugresistancebymagneticfe3o4nanoparticlecopolymerizatingdaunorubicinandmdr1shrnaexpressionvectorinleukemiacells
AT etal reversalofmultidrugresistancebymagneticfe3o4nanoparticlecopolymerizatingdaunorubicinandmdr1shrnaexpressionvectorinleukemiacells
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