Experimental Study on Knockdowning of Angiotensin-Converting Enzyme in Rat Vascular Endothelial Cells
Zhou Hua
Zhang Cuifan
Cheng Ruirui
Gao Fen
Abstract：Objective The adenovirus vector was constructed to carry the short hairpin RNA (shRNA) of angiotensin-converting enzyme (ACE),and the ACE expression was selectively knockdown in rat vascular endothelia1 cells (ECs) by RNA interference.Methods The segments ACE-shRNA was obtained from plasmid p-ACE-shRNA that was constructed at an earlier date,and then was cloned into the pDC316 shuttle plasmid to form the vector pDC316-EGFP-ACE-shRNA by RT-PCR.To package the recombinant adenovirus,the plasmid pDC316-EGFP-ACE-shRNA was cotransfected with pBHGlox-E1,3Cre genomic plasmid into 293 cells.The recombinant adenovirus was transfected into the primary cultured rat vascular ECs.ACE mRNA expression were analyzed from rat vascular ECs before transfection and 24 hours,48 hours,72 hours after transfection by real-time fluorescence quantitative PCR.Results Recombinant adenoviral vector Ad EGFP-ACE-shRNA was constructed successfully that was confirmed by restriction enzyme digestion,PCR and GFP expression.Expression of ACE mRNA in rat vascular ECs were significantly reduced 48 hours after Ad-EGFP-ACE-shRNA transfection,and was nearly undetectable after 72 hours.Conclusion The recombinant adenoviral vector which carry ACE-shRNA is successfully constructed.RNA interference can knockdown ACE expression in cultured ECs,which maybe provide a new gene therapy idea for cardiovascular diseases.
Keywords：RNA interferenceendothelial cellsengiotensin-converting enzyme
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 859-864 )