MENG Fan-ping,YANG Gang-yi,LI Ke,LI Ling,QI Xiao-ya,LIU Jian-lei,LI Rong.Effects of recombinant adenovirus-mediated siRNA on adiponectin expression in 3T3-L1 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(14):1312-1315.
MENG Fan-ping,YANG Gang-yi,LI Ke,LI Ling,QI Xiao-ya,LIU Jian-lei,LI Rong.Effects of recombinant adenovirus-mediated siRNA on adiponectin expression in 3T3-L1 cells[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(14):1312-1315.DOI:
Objective To construct the RNA interfered adenovirus expression vector specific for Adiponectin (Acrp30) gene and observe its effect on the Acrp30 expression in 3T3-L1 cells. Methods Firstly
fragments U6-Acrp30-1 and U6-Acrp30-3 were obtained by using KpnⅠ+NotⅠ to digest the plasmids pSilencer1.0-U6-Acrp30-1 and pSilencer1.0- U6-Acrp30-3. Two adenovirus vector plasmids
Ad-Acrp30-1 and Ad-Acrp30-3 were constructed according to a two-step transformation protocol. Two newly constructed plasmids were transfected into 293 packaging cells to produce adenovirus
which were further multiplied and purified. Then 3T3-L1 cells were infected with the 2 recombinant adenoviruses respectively. The mRNA expression and protein levels of Acrp30 in these cells were evaluated by semi-quantitative RT-PCR and ELISA. Results Two recombinant adenoviral plasmids Ad-Acrp30-1 and Ad-Acrp30-3 were successfully constructed. They-remarkably down-regulated the expression of Acrp30 at both the mRNA and protein levels in transfected 3T3-L1 cells. Conclusion The siRNA eukaryotic expression vectors against Acrp30 mRNA effectively inhibit the expression of Acrp30 in 3T3-L1 adipocytes.