title: Experimental endostatin-GFP gene transfection into human retinal vascular endothelial cells using ultrasound-targeted cationic microbubble destruction.
abstract: OBJECTIVE
  The purpose of this study was to investigate whether ultrasound-targeted cationic microbubble destruction could effectively deliver endostatin-green fluorescent protein (ES-GFP) plasmids to human retinal vascular endothelial cells (HRECs).
  
  
  METHODS
  Cationic microbubbles (CMBs) were prepared and then compared with neutral microbubbles (NMBs) and liposomes. First, the two types of microbubbles were characterized in terms of size and zeta potential. The cell viability of the HRECs was measured using the 3-(4,5-dimthylthiazol-2-yl)-2,5 diphenyl-tetrazolium bromide (MTT) assay. The transcription and expression of endostatin, VEGF, Bcl-2, and Bcl-xl were measured via quantitative real-time PCR (qPCR) and western blotting, respectively.
  
  
  RESULTS
  CMBs differed significantly from NMBs in terms of the zeta potential, but no differences in size were detected. Following ultrasound-targeted microbubble destruction (UTMD)-mediated gene therapy, the transcription and expression of endostatin were highest in the CMB group (p<0.05), while the transcription and expression of VEGF, Bcl-2, and Bcl-xl were lowest compared with the other groups. Moreover, the inhibition of HREC growth was enhanced following treatment with CMBs compared with NMBs or liposomes in vitro (p<0.01).
  
  
  CONCLUSIONS
  This study demonstrated that ultrasound-mediated cationic microbubbles could enhance the transfection efficiency of ES-GFP, which had obvious impacts on the inhibition of the growth process of HRECs in vitro. These results suggest that the combination of UTMD and ES-GFP compounds might be a useful tool for gene therapy targeting retinal neovascularization.
authors: Institute of Ultrasound Imaging and Department of Ultrasound, Second Affiliated Hospital of Chongqing Medical University; Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing, P.R. China.
authors: Yan
authors: Y
authors: Xu
authors: Department of Anesthesiology, Chongqing City Hospital of Traditional Chinese Medicine, Chongqing, P. R. China.
authors: Zongyuan
authors: Z
authors: Xie
authors: Institute of Ultrasound Imaging and Department of Ultrasound, Second Affiliated Hospital of Chongqing Medical University; Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing, P.R. China.
authors: Yu
authors: Y
authors: Zhou
authors: Department of Ophthalmology, Second Affiliated Hospital of Chongqing Medical University, Chongqing, P. R. China.
authors: Xiyuan
authors: X
authors: Zhou
authors: Institute of Ultrasound Imaging and Department of Ultrasound, Second Affiliated Hospital of Chongqing Medical University; Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing, P.R. China.
authors: Pan
authors: P
authors: Li
authors: Institute of Ultrasound Imaging and Department of Ultrasound, Second Affiliated Hospital of Chongqing Medical University; Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing, P.R. China.
authors: Zhigang
authors: Z
authors: Wang
authors: Institute of Ultrasound Imaging and Department of Ultrasound, Second Affiliated Hospital of Chongqing Medical University; Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing, P.R. China.
authors: Qunxia
authors: Q
authors: Zhang
mesh_terms: D002412:Cations; D049109:Cell Proliferation; D043169:Endostatins / Q000235:genetics* / Q000627:therapeutic use; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D045423:Microbubbles / Q000627:therapeutic use; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use; D015861:Retinal Neovascularization / Q000473:pathology / Q000628:therapy; D012171:Retinal Vessels / Q000166:cytology / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics
chemical_list: D002412:Cations; D043169:Endostatins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins