The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro

Ying Wu1, Weiwei Wang2, Yinting Chen1, Kaihong Huang1, Xintao Shuai2, Qikui Chen1, Xuexian Li1, Guoda Lian11Department of Gastroenterology, The Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China; 2BME Center, School of Chemistry and Chemical Engineering, Sun Yat-Sen University, G...

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Autores principales:	Ying Wu, Weiwei Wang, Yinting Chen, et al
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2010
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/e54a6d164bdf4d309b14f6b46d9cc794
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spelling	oai:doaj.org-article:e54a6d164bdf4d309b14f6b46d9cc7942021-12-02T08:07:38ZThe investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro1176-91141178-2013https://doaj.org/article/e54a6d164bdf4d309b14f6b46d9cc7942010-03-01T00:00:00Zhttp://www.dovepress.com/the-investigation-of-polymer-sirna-nanoparticle-for-gene-therapy-of-ga-a4041https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Ying Wu1, Weiwei Wang2, Yinting Chen1, Kaihong Huang1, Xintao Shuai2, Qikui Chen1, Xuexian Li1, Guoda Lian11Department of Gastroenterology, The Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China; 2BME Center, School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou, ChinaAbstract: Small interfering RNA (siRNA) molecules have significant therapeutic promise for the genetic treatment of cancer. To overcome instability and low transfection efficiency, polyethylene glycol-polyethyleneimine (PEG-PEI) was synthesized and investigated as a non-viral carrier of siRNA targeting CD44v6 in gastric carcinoma cells. The size, surface charge using zeta potential, and morphology via scanning electron microscopy (SEM) of PEG-PEI/siRNA nanoparticles was characterized, and their cytotoxicity, transfection efficiency, and interaction with SGC7901 human gastric carcinoma cells was evaluated. The transfection efficiency of PEG-PEI/siRNA nanocomplexes was dependant on the charge ratio between amino groups of PEG-PEI and phosphate groups of siRNA (N/P) values, which reflected the molar ratio of PEG-PEI to siRNA during complex formation. The transfection efficiency of PEG-PEI/siRNA at N/P 15 was 72.53% &plusmn; 2.38%, which was higher than that observed using Lipofectamine 2000 and PEI as delivery carriers. Cytotoxicity of PEG-PEI was determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and was obviously lower than that of PEI. Moreover, when N/P was below 15, PEG-PEI/siRNA was less toxic than Lipofectamine 2000/siRNA. RT-PCR (real time polymerase chain reaction) and Western blot analyses of CD44v6 expression demonstrated the gene silencing effect of PEG-PEI/siRNA at N/P 15. These data indicate that PEG-PEI may be a promising non-viral carrier for altering gene expression in the treatment of gastric cancer with many advantages, such as relatively high gene transfection efficiency and low cytotoxicity.Keywords: siRNA, PEG-PEI, nanoparticles, CD44v6 gene, gastric carcinoma cells Ying WuWeiwei WangYinting Chenet alDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2010, Iss default, Pp 129-136 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Ying Wu Weiwei Wang Yinting Chen et al The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
description	Ying Wu1, Weiwei Wang2, Yinting Chen1, Kaihong Huang1, Xintao Shuai2, Qikui Chen1, Xuexian Li1, Guoda Lian11Department of Gastroenterology, The Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China; 2BME Center, School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou, ChinaAbstract: Small interfering RNA (siRNA) molecules have significant therapeutic promise for the genetic treatment of cancer. To overcome instability and low transfection efficiency, polyethylene glycol-polyethyleneimine (PEG-PEI) was synthesized and investigated as a non-viral carrier of siRNA targeting CD44v6 in gastric carcinoma cells. The size, surface charge using zeta potential, and morphology via scanning electron microscopy (SEM) of PEG-PEI/siRNA nanoparticles was characterized, and their cytotoxicity, transfection efficiency, and interaction with SGC7901 human gastric carcinoma cells was evaluated. The transfection efficiency of PEG-PEI/siRNA nanocomplexes was dependant on the charge ratio between amino groups of PEG-PEI and phosphate groups of siRNA (N/P) values, which reflected the molar ratio of PEG-PEI to siRNA during complex formation. The transfection efficiency of PEG-PEI/siRNA at N/P 15 was 72.53% &plusmn; 2.38%, which was higher than that observed using Lipofectamine 2000 and PEI as delivery carriers. Cytotoxicity of PEG-PEI was determined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and was obviously lower than that of PEI. Moreover, when N/P was below 15, PEG-PEI/siRNA was less toxic than Lipofectamine 2000/siRNA. RT-PCR (real time polymerase chain reaction) and Western blot analyses of CD44v6 expression demonstrated the gene silencing effect of PEG-PEI/siRNA at N/P 15. These data indicate that PEG-PEI may be a promising non-viral carrier for altering gene expression in the treatment of gastric cancer with many advantages, such as relatively high gene transfection efficiency and low cytotoxicity.Keywords: siRNA, PEG-PEI, nanoparticles, CD44v6 gene, gastric carcinoma cells
format	article
author	Ying Wu Weiwei Wang Yinting Chen et al
author_facet	Ying Wu Weiwei Wang Yinting Chen et al
author_sort	Ying Wu
title	The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
title_short	The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
title_full	The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
title_fullStr	The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
title_full_unstemmed	The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro
title_sort	investigation of polymer-sirna nanoparticle for gene therapy of gastric cancer in vitro
publisher	Dove Medical Press
publishDate	2010
url	https://doaj.org/article/e54a6d164bdf4d309b14f6b46d9cc794
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The investigation of polymer-siRNA nanoparticle for gene therapy of gastric cancer in vitro

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