Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.

<h4>Background</h4>The BCRP/ABCG2 transporter, which mediates drug resistance in many types of cells, depends on energy provided by ATP hydrolysis. Here, a retrovirus encoding a shRNA targeting the ATP-binding domain of this protein was used to screen for highly efficient agents that cou...

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Autores principales:	Ni Xie, Lisha Mou, Jianhui Yuan, Wenlan Liu, Tingting Deng, Zigang Li, Yi Jing, Zhangli Hu
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/43a98db68abb48c88b579509a6b92559
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spelling	oai:doaj.org-article:43a98db68abb48c88b579509a6b925592021-11-25T06:06:39ZModulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.1932-620310.1371/journal.pone.0103463https://doaj.org/article/43a98db68abb48c88b579509a6b925592014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25076217/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>The BCRP/ABCG2 transporter, which mediates drug resistance in many types of cells, depends on energy provided by ATP hydrolysis. Here, a retrovirus encoding a shRNA targeting the ATP-binding domain of this protein was used to screen for highly efficient agents that could reverse drug resistance and improve cell sensitivity to drugs, thus laying the foundation for further studies and applications.<h4>Methodology/principal findings</h4>To target the ATP-binding domain of BCRP/ABCG2, pLenti6/BCRPsi shRNA recombinant retroviruses, with 20 bp target sequences starting from the 270th, 745th and 939th bps of the 6th exon, were constructed and packaged. The pLenti6/BCRPsi retroviruses (V-BCRPi) that conferred significant knockdown effects were screened using a drug-sensitivity experiment and flow cytometry. The human choriocarcinoma cell line JAR, which highly expresses endogenous BCRP/ABCG2, was injected under the dorsal skin of a hairless mouse to initiate a JAR cytoma. After injecting V-BCRPi-infected JAR tumor cells into the dorsal skin of hairless mice, BCRP/ABCG2 expression in the tumor tissue was determined using immunohistochemistry, fluorescent quantitative RT-PCR and Western blot analyses. After intraperitoneal injection of BCRP/ABCG2-tolerant 5-FU, the tumor volume, weight change, and apoptosis rate of the tumor tissue were determined using in situ hybridization. V-BCRPi increased the sensitivity of the tumor histiocytes to 5-FU and improved the cell apoptosis-promoting effects of 5-FU in the tumor.<h4>Conclusions/significance</h4>The goal of the in vivo and in vitro studies was to screen for an RNA interference recombinant retrovirus capable of stably targeting the ATP-binding domain of BCRP/ABCG2 (V-BCRPi) to inhibit its function. A new method to improve the chemo-sensitivity of breast cancer and other tumor cells was discovered, and this method could be used for gene therapy and functional studies of malignant tumors.Ni XieLisha MouJianhui YuanWenlan LiuTingting DengZigang LiYi JingZhangli HuPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 7, p e103463 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ni Xie Lisha Mou Jianhui Yuan Wenlan Liu Tingting Deng Zigang Li Yi Jing Zhangli Hu Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
description	<h4>Background</h4>The BCRP/ABCG2 transporter, which mediates drug resistance in many types of cells, depends on energy provided by ATP hydrolysis. Here, a retrovirus encoding a shRNA targeting the ATP-binding domain of this protein was used to screen for highly efficient agents that could reverse drug resistance and improve cell sensitivity to drugs, thus laying the foundation for further studies and applications.<h4>Methodology/principal findings</h4>To target the ATP-binding domain of BCRP/ABCG2, pLenti6/BCRPsi shRNA recombinant retroviruses, with 20 bp target sequences starting from the 270th, 745th and 939th bps of the 6th exon, were constructed and packaged. The pLenti6/BCRPsi retroviruses (V-BCRPi) that conferred significant knockdown effects were screened using a drug-sensitivity experiment and flow cytometry. The human choriocarcinoma cell line JAR, which highly expresses endogenous BCRP/ABCG2, was injected under the dorsal skin of a hairless mouse to initiate a JAR cytoma. After injecting V-BCRPi-infected JAR tumor cells into the dorsal skin of hairless mice, BCRP/ABCG2 expression in the tumor tissue was determined using immunohistochemistry, fluorescent quantitative RT-PCR and Western blot analyses. After intraperitoneal injection of BCRP/ABCG2-tolerant 5-FU, the tumor volume, weight change, and apoptosis rate of the tumor tissue were determined using in situ hybridization. V-BCRPi increased the sensitivity of the tumor histiocytes to 5-FU and improved the cell apoptosis-promoting effects of 5-FU in the tumor.<h4>Conclusions/significance</h4>The goal of the in vivo and in vitro studies was to screen for an RNA interference recombinant retrovirus capable of stably targeting the ATP-binding domain of BCRP/ABCG2 (V-BCRPi) to inhibit its function. A new method to improve the chemo-sensitivity of breast cancer and other tumor cells was discovered, and this method could be used for gene therapy and functional studies of malignant tumors.
format	article
author	Ni Xie Lisha Mou Jianhui Yuan Wenlan Liu Tingting Deng Zigang Li Yi Jing Zhangli Hu
author_facet	Ni Xie Lisha Mou Jianhui Yuan Wenlan Liu Tingting Deng Zigang Li Yi Jing Zhangli Hu
author_sort	Ni Xie
title	Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
title_short	Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
title_full	Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
title_fullStr	Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
title_full_unstemmed	Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.
title_sort	modulating drug resistance by targeting bcrp/abcg2 using retrovirus-mediated rna interference.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/43a98db68abb48c88b579509a6b92559
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_version_	1718414157761478656

Modulating drug resistance by targeting BCRP/ABCG2 using retrovirus-mediated RNA interference.

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