Expression of Reverse
Cholesterol Transport Pathway Associated Protein Genes in Skeletal Muscle
FAN Le-Ming, ZHANG Hui, CHEN Qi, WEI En-Hui, CAI Hai-Jiang
( Atherosclerosis Research Center, Nanjing Medical University, Nanjing 210029,
China )
Abstract Viral and
nonviral vectors containing apoAI, apoE or LCAT
genes were constructed and transfected into myogenic cells in vitro or
injected directly into mouse skeletal muscle. The expression efficiencies of
these vectors were assaied to investigate the possibility of ectopic expression
of these genes in skeletal muscle and to develop a safe and convenient gene
therapy method for atherosclerosis. The results showed that the primary
cultured mouse myoblasts, C2C12 cells transfected with pCMVapoE3
expressed human apoE3 successfully and the expressed product was secreted into
the medium. Mouse skeletal muscle efficiently expressed apoE3 in vivo
after direct plasmid injection. The expression level of Ad-RSV-apoAI
in primary cultured mouse myoblasts was correlated with virus titer. Human
apoAI was synthesized in mouse skeletal muscle by direct injection of
recombinant virus and was secreted into blood continuously up to 30 days； functional LCAT was expressed
by C2C12 and 293 cells transfected with conventional vector or recombinant AAV
plasmid DNA. The expression efficiency of recombinant AAV plasmid DNA was 2―5
times higher than that of conventional plasmid vector. The above results
provided experimental data for further studying and developing a gene therapy
method for atherosclerosis by enhancement of reverse cholesterol transport
using skeletal muscle as target.
Key words apolipoprotein AI； apolipoprotein E； lecithin-cholesterol
acyltransferase； reverse cholesterol transport； gene transfer
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