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Ultrasound-targeted microbubble destruction (UTMD) assisted delivery of shRNA against PHD2 into H9C2 cells
Journal article   Open access   Peer reviewed

Ultrasound-targeted microbubble destruction (UTMD) assisted delivery of shRNA against PHD2 into H9C2 cells

Li Zhang, Zhenxing Sun, Pingping Ren, Robert J Lee, Guangya Xiang, Qing Lv, Wei Han, Jing Wang, Shuping Ge and Mingxing Xie
PloS one, v 10(8), pp e0134629-e0134629
2015
DOI: https://doi.org/10.1371/journal.pone.0134629
PMID: 26267649
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1371/journal.pone.0134629View
Published, Version of Record (VoR)CC BY V4.0,  Open

Abstract

Cell Line Gene Expression - genetics Gene Transfer Techniques Genetic Therapy Green Fluorescent Proteins - genetics Humans Hypoxia-Inducible Factor-Proline Dioxygenases - antagonists & inhibitors Hypoxia-Inducible Factor-Proline Dioxygenases - biosynthesis Hypoxia-Inducible Factor-Proline Dioxygenases - genetics Microbubbles Plasmids - genetics Polyethyleneimine - administration & dosage Polyethyleneimine - chemistry RNA, Small Interfering - administration & dosage RNA, Small Interfering - genetics Transforming Growth Factor beta - genetics Ultrasonics Transfection
Gene therapy has great potential for human diseases. Development of efficient delivery systems is critical to its clinical translation. Recent studies have shown that microbubbles in combination with ultrasound (US) can be used to facilitate gene delivery. An aim of this study is to investigate whether the combination of US-targeted microbubble destruction (UTMD) and polyethylenimine (PEI) (UTMD/PEI) can mediate even greater gene transfection efficiency than UTMD alone and to optimize ultrasonic irradiation parameters. Another aim of this study is to investigate the biological effects of PHD2-shRNA after its transfection into H9C2 cells. pEGFP-N1 or eukaryotic shPHD2-EGFP plasmid was mixed with albumin-coated microbubbles and PEI to form complexes for transfection. After these were added into H9C2 cells, the cells were exposed to US with various sets of parameters. The cells were then harvested and analyzed for gene expression. UTMD/PEI was shown to be highly efficient in gene transfection. An US intensity of 1.5 W/cm2, a microbubble concentration of 300μl/ml, an exposure time of 45s, and a plasmid concentration of 15μg/ml were found to be optimal for transfection. UTMD/PEI-mediated PHD2-shRNA transfection in H9C2 cells significantly down regulated the expression of PHD2 and increased expression of HIF-1α and downstream angiogenesis factors VEGF, TGF-β and bFGF. UTMD/PEI, combined with albumin-coated microbubbles, warrants further investigation for therapeutic gene delivery.

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#3 Good Health and Well-Being

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Domestic collaboration
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Web of Science research areas
Pharmacology & Pharmacy
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