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Title: Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG{sub 2} cells

Abstract

Ultrasound-targeted microbubble destruction had been employed in gene delivery and promised great potential. Liver has unique features that make it attractive for gene therapy. However, it poses formidable obstacles to hepatocyte-specific gene delivery. This study was designed to test the efficiency of therapeutic gene transfer and expression mediated by ultrasound/microbubble strategy in HepG{sub 2} cell line. Air-filled albumin microbubbles were prepared and mixed with plasmid DNA encoding low density lipoprotein receptor (LDLR) and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured HepG{sub 2} cells under variable ultrasound conditions. Transfection rate of the transferred gene and cell viability were assessed by FACS analysis, confocal laser scanning microscopy, Western blot analysis and Trypan blue staining. The result demonstrated that microbubbles with ultrasound irradiation can significantly elevate exogenous LDLR gene expression and the expressed LDLRs were functional and active to uptake their ligands. We conclude that ultrasound-targeted microbubble destruction has the potential to promote safe and efficient LDLR gene transfer into hepatocytes. With further refinement, it may represent an effective nonviral avenue of gene therapy for liver-involved genetic diseases.

Authors:
 [1];  [2];  [2];  [3];  [3];  [3];  [3];  [1];  [2];  [2];  [4];  [5];  [5];  [5]
  1. Institute of Reproductive Medicine, Nanjing Medical University, Nanjing 210029 (China)
  2. (China)
  3. Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029 (China)
  4. Institute of Reproductive Medicine, Nanjing Medical University, Nanjing 210029 (China) and Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029 (China) and Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029 (China). E-mail: lmfan@njmu.edu.cn
  5. Cardiovascular Division, Wuxi No.1 Hospital, Wuxi 214002 (China)
Publication Date:
OSTI Identifier:
20798932
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 343; Journal Issue: 2; Other Information: DOI: 10.1016/j.bbrc.2006.02.179; PII: S0006-291X(06)00480-3; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ALBUMINS; GENE THERAPY; GENES; HEREDITARY DISEASES; IRRADIATION; LIGANDS; LIPOPROTEINS; LIVER; LIVER CELLS; MICROSCOPY; RECEPTORS; TRYPAN BLUE; UPTAKE

Citation Formats

Guo Dongping, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Li Xiaoyu, Sun, Ping, Tang Yibo, Chen Xiuying, Chen Qi, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Fan Leming, Zang Bin, Shao Lizheng, and Li Xiaorong. Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG{sub 2} cells. United States: N. p., 2006. Web.
Guo Dongping, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Li Xiaoyu, Sun, Ping, Tang Yibo, Chen Xiuying, Chen Qi, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Fan Leming, Zang Bin, Shao Lizheng, & Li Xiaorong. Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG{sub 2} cells. United States.
Guo Dongping, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Li Xiaoyu, Sun, Ping, Tang Yibo, Chen Xiuying, Chen Qi, Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029, Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029, Fan Leming, Zang Bin, Shao Lizheng, and Li Xiaorong. Fri . "Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG{sub 2} cells". United States. doi:.
@article{osti_20798932,
title = {Ultrasound-targeted microbubble destruction improves the low density lipoprotein receptor gene expression in HepG{sub 2} cells},
author = {Guo Dongping and Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029 and Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029 and Li Xiaoyu and Sun, Ping and Tang Yibo and Chen Xiuying and Chen Qi and Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029 and Provincial Laboratory of Human Functional Genomics, Nanjing Medical University, Nanjing 210029 and Fan Leming and Zang Bin and Shao Lizheng and Li Xiaorong},
abstractNote = {Ultrasound-targeted microbubble destruction had been employed in gene delivery and promised great potential. Liver has unique features that make it attractive for gene therapy. However, it poses formidable obstacles to hepatocyte-specific gene delivery. This study was designed to test the efficiency of therapeutic gene transfer and expression mediated by ultrasound/microbubble strategy in HepG{sub 2} cell line. Air-filled albumin microbubbles were prepared and mixed with plasmid DNA encoding low density lipoprotein receptor (LDLR) and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured HepG{sub 2} cells under variable ultrasound conditions. Transfection rate of the transferred gene and cell viability were assessed by FACS analysis, confocal laser scanning microscopy, Western blot analysis and Trypan blue staining. The result demonstrated that microbubbles with ultrasound irradiation can significantly elevate exogenous LDLR gene expression and the expressed LDLRs were functional and active to uptake their ligands. We conclude that ultrasound-targeted microbubble destruction has the potential to promote safe and efficient LDLR gene transfer into hepatocytes. With further refinement, it may represent an effective nonviral avenue of gene therapy for liver-involved genetic diseases.},
doi = {},
journal = {Biochemical and Biophysical Research Communications},
number = 2,
volume = 343,
place = {United States},
year = {Fri May 05 00:00:00 EDT 2006},
month = {Fri May 05 00:00:00 EDT 2006}
}