Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line

Pardis Mokhtary,1,2 Bita Javan,1,3 Mahrokh Sharbatkhari,4 Alireza Soltani,5 Vahid Erfani-Moghadam1,2 1Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran; 2Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan Un...

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Autores principales:	Mokhtary P, Javan B, Sharbatkhari M, Soltani A, Erfani-Moghadam V
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2018
Materias:	Cationic vehicles shRNA UCA1 Apoptosis Gene delivery Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/5e442412b05746f6941ed03e21768cbc
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spelling	oai:doaj.org-article:5e442412b05746f6941ed03e21768cbc2021-12-02T07:24:27ZCationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line1178-2013https://doaj.org/article/5e442412b05746f6941ed03e21768cbc2018-11-01T00:00:00Zhttps://www.dovepress.com/cationic-vesicles-for-efficient-shrna-transfection-in-the-mcf-7-breast-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Pardis Mokhtary,1,2 Bita Javan,1,3 Mahrokh Sharbatkhari,4 Alireza Soltani,5 Vahid Erfani-Moghadam1,2 1Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran; 2Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran; 3Department of Molecular Medicine, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran; 4R&D Section, Arya Tina Gene Biopharmaceutical Company, Gorgan, Iran; 5Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Introduction: Novel and safe delivery solutions for RNAi therapeutics are essential to obtain the full potential of cancer gene therapy. Methods: In this study, cationic vesicular nanocarrier was applied for delivering lnc urothelial carcinoma-associated 1 (lnc UCA1) shRNA expression vector to MCF-7 cells. The physicochemical characteristics, cytotoxicity, and transfection efficiency of cationic vesicles prepared from various molar ratios of amphiphilic surfactant Tween 80 (T), squalene (S), cationic charge lipid didodecyldimethylammonium bromide, and polyethylenimine were investigated. The particle sizes of the vesicles in the nanosize range were determined by dynamic light scattering and transmission electron microscopy. Results: Gel protection assay with agarose gel electrophoresis showed cationic vesicles can protect the shRNA plasmid from DNase 1 enzyme. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt result showed no significant cytotoxicity was caused in MCF-7 cancer cell line by (T:S):polyethylenimine cationic vesicles. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt assay, fluorescence microscope images, and flow cytometry analyses confirmed that (T:S)1,040 µM with 4.3 µg/mL of PEI vesicles provided effective transfection without significant cytotoxicity. Furthermore, we found efficient UCA1 shRNA transfection and significant (P<0.05) cell cycle arrest and apoptosis in MCF-7 cancer cells. Conclusion: The novel nonviral vesicular nanocarrier, (T:S)1,040 µM with 4.3 µg/mL of PEI, might be safe and efficient for cancer gene therapy and can be used in further in vitro and in vivo studies. Keywords: cationic vesicles, shRNA, UCA1, apoptosis, gene deliveryMokhtary PJavan BSharbatkhari MSoltani AErfani-Moghadam VDove Medical PressarticleCationic vehiclesshRNAUCA1ApoptosisGene deliveryMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 13, Pp 7107-7121 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cationic vehicles shRNA UCA1 Apoptosis Gene delivery Medicine (General) R5-920
spellingShingle	Cationic vehicles shRNA UCA1 Apoptosis Gene delivery Medicine (General) R5-920 Mokhtary P Javan B Sharbatkhari M Soltani A Erfani-Moghadam V Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
description	Pardis Mokhtary,1,2 Bita Javan,1,3 Mahrokh Sharbatkhari,4 Alireza Soltani,5 Vahid Erfani-Moghadam1,2 1Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran; 2Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran; 3Department of Molecular Medicine, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran; 4R&D Section, Arya Tina Gene Biopharmaceutical Company, Gorgan, Iran; 5Golestan Rheumatology Research Center, Golestan University of Medical Sciences, Gorgan, Iran Introduction: Novel and safe delivery solutions for RNAi therapeutics are essential to obtain the full potential of cancer gene therapy. Methods: In this study, cationic vesicular nanocarrier was applied for delivering lnc urothelial carcinoma-associated 1 (lnc UCA1) shRNA expression vector to MCF-7 cells. The physicochemical characteristics, cytotoxicity, and transfection efficiency of cationic vesicles prepared from various molar ratios of amphiphilic surfactant Tween 80 (T), squalene (S), cationic charge lipid didodecyldimethylammonium bromide, and polyethylenimine were investigated. The particle sizes of the vesicles in the nanosize range were determined by dynamic light scattering and transmission electron microscopy. Results: Gel protection assay with agarose gel electrophoresis showed cationic vesicles can protect the shRNA plasmid from DNase 1 enzyme. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt result showed no significant cytotoxicity was caused in MCF-7 cancer cell line by (T:S):polyethylenimine cationic vesicles. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium, inner salt assay, fluorescence microscope images, and flow cytometry analyses confirmed that (T:S)1,040 µM with 4.3 µg/mL of PEI vesicles provided effective transfection without significant cytotoxicity. Furthermore, we found efficient UCA1 shRNA transfection and significant (P<0.05) cell cycle arrest and apoptosis in MCF-7 cancer cells. Conclusion: The novel nonviral vesicular nanocarrier, (T:S)1,040 µM with 4.3 µg/mL of PEI, might be safe and efficient for cancer gene therapy and can be used in further in vitro and in vivo studies. Keywords: cationic vesicles, shRNA, UCA1, apoptosis, gene delivery
format	article
author	Mokhtary P Javan B Sharbatkhari M Soltani A Erfani-Moghadam V
author_facet	Mokhtary P Javan B Sharbatkhari M Soltani A Erfani-Moghadam V
author_sort	Mokhtary P
title	Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
title_short	Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
title_full	Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
title_fullStr	Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
title_full_unstemmed	Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line
title_sort	cationic vesicles for efficient shrna transfection in the mcf-7 breast cancer cell line
publisher	Dove Medical Press
publishDate	2018
url	https://doaj.org/article/5e442412b05746f6941ed03e21768cbc
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_version_	1718399457728397312

Cationic vesicles for efficient shRNA transfection in the MCF-7 breast cancer cell line

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