Hypertension, Vol 21, 894-899, Copyright © 1993 by American Heart Association
R Morishita, GH Gibbons, Y Kaneda, T Ogihara and VJ Dzau
Although many in vitro gene transfer methods already exist, such as calcium
phosphate precipitation, electroporation, or cationic liposomes, these
methods cause significant cell injury and cell death. The study of the
biology of endogenous autocrine-paracrine vasoactive systems such as the
renin-angiotensin system in vascular cells is limited by the lack of a
suitable gene transfer method with high efficiency of transfection and
expression that will permit cell biology studies. Recently, the Sendai
virus (hemagglutinating virus of Japan, HVJ)-liposome-mediated gene
transfer method has been shown to be an efficient and nontoxic method of
gene transfer. In this study, we characterized the efficiency and
suitability of the HVJ method for vascular biology research. Using SV40
T-antigen complementary DNA (cDNA), we initially compared the efficiency of
the HVJ method and lipofection for transfection of cultured vascular smooth
muscle cells (VSMCs). We observed that after 35 minutes of incubation, the
HVJ method exhibited a 10-fold higher efficiency of transfection than
lipofection. We used this method to study vascular angiotensin converting
enzyme (ACE) expression in cultured VSMCs and cultured rat carotid arteries
in vitro. The HVJ method of transfection of human ACE cDNA into VSMCs and
COS cells was significantly more efficient than lipofection. Using this
method, we demonstrated that transfection of ACE cDNA resulted in increased
DNA synthesis, which was inhibited by the specific angiotensin II receptor
antagonist DuP 753 (10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS)
ARTICLES
Novel in vitro gene transfer method for study of local modulators in vascular smooth muscle cells
Division of Cardiovascular Medicine, Falk Cardiovascular Research Center, Stanford University School of Medicine, Calif. 94305-5246.
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