Aim To investigate the possibility of ectopic expression of apolipoprotein AI (apo AI) and lecithin cholesterol acyltrasferase(LCAT) by myogenic cells and developing a new approach of gene therapy for atherosclerosis (As). Methods Recombinant replication-deficient viral particles were prepared with polycistronic retrovirus vectors containing apoAI cDNA, LCAT cDNA and the neomycine phosphotransferase gene (NEO), Mice primary cultured myoblasts and myogenic cell line C₂C₁₂ were transfected by these viruses. The efficiency of transfection and the state of integration were detected by PCR, while the expression of apo AI and LCAT were measured by ELISA and immunohistochemical method. Results All transfected mice myoblasts and C₂C₁₂ cells gained the ability of co-expressing human apo AI and LCAT. Stable transfected C₂C₁₂ cell line selected by G418 maintained the ability of co-expressing apo AI and LCAT for 60 days. PCR shown the apo AI cDNA and IRES sequence were integrated into genomes of target cells effectively. Conclusions These finds indicated mouse primary myoblasts and C₂C₁₂ myoblasts transduced with recombinant retroviral vectors could efficiently express and secrete human apo AI and LCAT. It suggested that the use of polycistronic retrovirus vectors containing human apo AI and LCAT cDNA to genetically modify myoblasts in vitro and then implantation back to skeletal muscle to high efficiently, long-term express apo AI and LCAT in vivo, might be a strategy to prevent or treat hypercholesterolemia and As by promoting the "reverse cholesterol transport" pathway.